Triple Infection Research Articles

Phylogenetic studies of bacteria (Rickettsia, Coxiella, and Anaplasma) in Amblyomma and Dermacentor ticks in Thailand and their co-infection

In this study, we attempted to detect Rickettsia, Coxiella and Anaplasma bacteria in one hundred and fourteen-Dermacentor and thirty three-Amblyomma unfed adult ticks that were collected from under leaves along animal trails at different places across Thailand. PCR amplification was used to identify bacterial infection with general conserved sequences of bacteria. The results revealed single infection in Amblyomma testudinarium ticks with Rickettsia (24%) and Coxiella (6%). Anaplasma bacteria were often detected in Dermacentor auratus ticks (32%). Coxiella spp. were detected in Dermacentor atrosignatus (6%) and D. auratus ticks (3%) in this study. Moreover, we found co-infection by Coxiella and Rickettsia bacteria (39%) in Am. testudinarium. In contrast, D. atrosignatus ticks were co-infected with Coxiella and Anaplasma bacteria (3%) and Dermacentor compactus ticks were co-infected with Rickettsia and Anaplasma spp. (25%). Interestingly, Am. testudinarium ticks (12%) were found for the first time to exhibit triple infection by these three bacteria. Phylogenetic studies showed the rickettsiae from ticks causing both single and multiple infections had sequence similarity with spotted fever group rickettsial strains, including Rickettsia massilliae, R. raoultii and R. tamurae. In addition, the phylogenetic analysis of the 16S rRNA gene of Coxiella bacteria showed that they were closely grouped with Coxiella endosymbionts in both Dermacentor and Amblyomma. Moreover, the Anaplasma identified in a D. auratus tick was grouped in the same clade with the pathogenic bacterium Anaplasma phagocytophilum. Bacterial co-infections in Dermacentor and Amblyomma ticks may cause co-transmission of some tick-borne microorganisms (pathogen and endosymbiont, whether enhance or reduce) in humans and animals and they could affect medical and veterinary health.

Infection by HTLV-1 Is Associated With High Levels of Proinflammatory Cytokines in HIV-HCV–Coinfected Patients

HIV, hepatitis C virus (HCV), and human T-cell lymphotropic virus type 1 (HTLV-1) share the same routes of infection, making coinfection by these viruses a frequent finding in endemic areas. However, there is scarce information on the clinical/immunological consequences of triple infection. Coinfection by HTLV-1 is able to modulate cytokine's production in patients with HIV, but there are no data on the immune response of HIV-HCV-HTLV-1-infected patients. We compared the plasma levels of 25 different cytokines in patients with HIV-HCV, according to their serostatus to HTLV-1 infection. Eligible patients should be on stable highly active antiretroviral therapy and have undetectable HIV-1 plasma viral load for, at least, 12 months. Cytokines levels were also evaluated by CD4 cells count, rates of sustained virological response (SVR) to previous HCV treatment, frequency of spontaneous HCV clearance, and HCV/IFN-λ3 genotypes. Twenty-five patients (15 coinfected by HIV and HCV, 10 coinfected by HIV, HCV, and HTLV-1) were evaluated. Among the triply infected group, 3 had undetectable HCV viremia (spontaneous clearance). All but one remaining patients were previously treated for HCV, with similar SVR rates (∼29%). Cytokines levels did not differ per HCV/IFN-λ3 genotypes, mean CD4 cells count, age, sex, or SVR. However, patients coinfected by HTLV-1 showed significantly higher levels of IL-1b, IL-2, TNF-α, IFN-γ, MIP-1α, RANTES, and interferon-induced protein 10 (IP-10) than HIV-HCV-coinfected ones. Patients presenting HCV spontaneous clearance had the highest levels of cytokines. Coinfection by HTLV-1 increases the plasma levels of proinflammatory cytokines of patients with HIV-HCV and can influence the outcomes of coinfected patients.

Evaluation of efficacy of serological methods for detection of HCV infection in blood donors: A single centre experience.

Background and Objective:Blood transfusion is an essential and life-saving medical intervention. Despite multiple preventive measures transfusion-transmitted hepatitis C virus (HCV) infection continues to be a major healthcare issue in Pakistan. This study was conducted at National Institute of Blood Diseases & Bone Marrow Transplantation to evaluate the frequency of active HCV infection with or without co-infection in blood donors and also to determine comparative efficacy of Multisure HCV antibody assay (MHAA); a new serological device.Methods:A total of 14652 blood donors visiting National Institute of Blood Diseases & Bone Marrow Transplantation (NIBD) Blood Bank from January 2013 to July 2014 were enrolled and screened for a range of blood borne infections such as HBV, HCV, HIV, malaria and syphilis. The HCV was screened simultaneously by Abbot Architect anti-HCV assay (CLIA) and MHAA. The active HCV infection was confirmed by nucleic acid testing (NAT) in reactive donors. Later; for determination of comparative efficacy of MHAA; all NAT positive samples were further tested using Monolisa™, HCV blot 3.0, Anti-HCV plus V2 and Anti-HCV-MPBIO-EIA.Results:The HCV reactive sera were observed in 1.563% (226) donors. The NAT confirmed active HCV infection in 138 donors. Overall 27.84% of HCV positive donors exhibited co-infection either with HBV (2.57%), syphilis (22.78%). Triple infection was not observed in any donor. The efficacy of MHAA is comparable to all the serological tests with a sensitivity of about 96.89%.Conclusion:Active HCV infection was present in 0.94% donors. With a sensitivity of 96.89% (95% CI: 95.66-98.12) the multi-parametric device MHAA can effectively detect HCV infection in donors. Thus, it can be used in limited health care settings for HCV screening.

Effect of some factors on intestinal parasitic infections

The present study has included collection of 1001 fecal samples of children aged seven years old and less than that age who visited different hospitals in Thi – Qar Province and in different area to ensure study for all province during the period from the first of October 2010 to the end of July 2011 to find out extent of the relationship between infection with intestinal parasites and factors age, sex, month, nature of water used for drinking, economic status, Location of residence, type of feeding , Education level of parents and Signs or clinical symptoms associated with infection of intestinal parasites . The fecal samples were examined by two methods: direct and indirect by floatation . The results have showed that percentage of infected children with parasites was % 33.3 . The single infections were prevailing with % 32 and Entamoeba histolytica formed high percentage % 28.8 in it , while the double infections reached % 1.2 as for the triple infection was found in one case in male aged four years old . Significant differences weren't found between intestinal parasitic infection and factors age , six , months, location of residence, type of feeding and educational level of parents while the differences were significant for economic factor and the water used for drinking and Signs or clinical symptoms associated with infection of intestinal parasites .

Detection and identification of blood-borne infections in dogs in Nigeria using light microscopy and the polymerase chain reaction.

Many sick dogs brought to the University of Ibadan Veterinary Teaching Hospital (UIVTH) are infested with ticks and are anemic. Up until recently, light microscopy (LM) has been the only available means used for detection of blood-borne infections. In other parts of the world, PCR-based assays been used as a gold standard for accurate diagnosis of blood-borne infections. In this study, we used LM and broad-spectrum rRNA gene PCR-based assays on 116 blood samples from dogs brought to the UIVTH for detection of the 18S rRNA gene of Babesia and the 16S rRNA genes of Ehrlichia and hemotropic mycoplasmas. The relationship between clinicopathological findings and PCR results was evaluated. Age, sex, presence of ticks, anemia, co-infection status, and fever were also assessed in relation to PCR positivity to determine the risk factors using stepwise logistic regression analyses. Light microscopic examination revealed an overall prevalence of infection of 14.7% (17/116). Organisms detected were Babesia canis (3.5%), Ehrlichia canis (10.3%) and Trypanosoma congolense (0.9%) and a single co-infection with Babesia canis and Ehrlichia canis (0.9%). PCR analysis revealed 89/116 (76.7%) positive samples. Infections with 1, 2 and 3 infectious agents occurred in 49 (55.1%), 36 (40.4%) and 4 (4.5%) samples, respectively. Specifically, among the 89 PCR positive samples, Babesia spp. (85.4%) was the most abundant infection followed by Ehrlichia spp. (46.1%) and hemoplasmas (13.5%). Sequencing of PCR products identified two samples (1.7%) that contained Hepatozoon canis DNA. Sequencing of hemoplasma positive samples identified 'Candidatus Mycoplasma haemobos' in 0.8% of dogs. Using PCR, a 5-fold higher prevalence of blood-borne infections was found in the dogs (76.7%, 89/116) than with LM (14.7%, 17/116) alone" Dogs between 1 and 12months were the most frequently infected with multiple agents (47.2% double and 50.0% triple infections). Male dogs had the highest prevalence of infection (80.4%) and more triple infections (75.0%). A total of 57.3% of infected dogs were anemic. Anemic dogs were 2.77 times more likely to test positive for Ehrlichia spp. (OR: 2.77 95% CI: 1.25-6.16) and dogs with ticks were 3.6 times more likely to test positive for hemoplasmas (OR=3.60 95% CI: 1.05-12.38). This study underscores the abundance of blood-borne infections in dogs in Ibadan, Nigeria, which is underestimated using light microscopy. This is also the first evidence of existence of 'Candidatus Mycoplasma haemobos' in a dog in Nigeria and in Africa. Consequently there is a need for molecular diagnostic facilities for routine screening of sick animals, as multiple infections were not found by light microscopy.

Etiological structure of intrauterine infections in pregnant and newborns with a complicated course of the early neonatal period

Aim of research – the study of the etiological structure of intrauterine infection in pregnant and estimation of their influence on the early neonatal period.Materials and methods. The material for the study was venous blood of pregnant, umbilical blood, breast maternal milk, mother’s saliva, newborn’s saliva of 114 pairs “mother-newborn”, divided in two groups – control one of physiological course of pregnancy, childbirth and early neonatal period and the main one with clinical symptoms of IUI. The diagnosis was proved by laboratory serological examinations for determining concentrations of IgM to Mycoplasma hominis, Chlamydia trahomatis, Ureaplasma urealyticum, to the herpes simplex virus and concentration and avidity index of specific IgG to HSV, to the virus of 6 type herpes, to the cytomegalovirus. The cytoscopic method was used together with serological ones to diagnose the cytomegaloviral infection.Research results. The increased concentration of IgM antibodies to M. hominis was revealed in the biological material (venous and umbilical blood) of pairs “woman-newborn) with the complicated course of the neonatal period in 9,1 % of examined persons, in 45,4 % of pairs – IgM to U. urealyticum, and in 81,8 % -to Ch. Trahomatis. IgM to the virus of herpes simplex virus (HVS) were revealed in 19,35 % of samples of venous blood, in 9,67 % of saliva samples and in 6,45 % of samples of breast milk of women; and IgM concentrations to HVS were (0,342±0,06) IU/ml, (0,117±0,04) IU/ml) and (0,438±0,001) IU/ml, respectively. Most samples of the biomaterial included low-avid IgG antibodies to the herpes simplex virus, 76 % of samples of women’s venous blood, 64 % of samples of breast milk and 38,7 % contained low-avid IgG to 6 type herpes virus. The cytomegaloviral infection in this group was diagnosed in 24 % of women and 32 % of newborns. The associated bacterial and viral infection were revealed in 65 % of examined pairs “mother-newborn” with the complicated course of the neonatal period, the triple infection with chlamydias, herpes simplex virus and cytomegalovirus was noted in 14 %; the association of herpes simplex virus, cytomegalovirus and ureaplasma was detected in 5 %. The double chamydia and herpes viral infection of 6 type were observed in 15 % of examined persons, at that these infections were added with ureoplasma in 7 %, with mycoplasma – in 5 %. The mix-infection of urogenital agents – chlamydias and ureaplasma was associated with the cytomegalovirus in 4 % of cases.Conclusions1. As a result of the realized study, the diagnosis was confirmed in all newborns with a suspected intrauterine infection.2. Mix-infections – Chlamydia, associated with viruses of the herpetic group occupy the leading place in the etiological structure of IUI agents, and the neonatal period is most complicated in children with double or triple infection.3. The leading clinical symptoms in children with IUI are the intrauterine development delay, intrauterine pneumonia, conjugated icterus, conjunctivitis

Prevalence of Human Papillomavirus in Women from the State of Michoacan, Mexico, Showed High Frequency of Unusual Virus Genotypes.

Human papillomaviruses (HPVs), the leading cause of cervical cancer, are distributed worldwide, with high prevalence in developing countries. The objective of the study is to know the prevalence and genotypes of HPV in women from the state of Michoacán and the Women's Hospital in Morelia, Michoacán. Cervical smear samples (159,288) were subjected to HPV detection by hybrid capture 2. A subsample of 484 patients from the Women's Hospital was studied by Papanicolaou test and linear array HPV genotyping, and when positive, patients were also examined by colposcopy and histopathology. The overall prevalence for HPV in Michoacán State was 7.74%; 7.11% in 2009, 6.46% in 2010, 9.58% in 2011, and 8.43% in 2012. The highest prevalence was found in the age groups < 25 and 25-34 years. The prevalence at the Women's Hospital was 8.51%. Cytological examination revealed normal cytology in 64.44% of samples, 26.66 % with low-grade and 8.88 % with high-grade squamous intraepithelial lesion (HSIL). However, by colposcopy, normal tissue appearance was found only in 26.66%; 51% were reclassified as low-grade squamous intraepithelial lesion, 17.77% as HSIL, and in 4.4% atrophy was observed. The most prevalent genotype in single infections was HPV59, followed by HPV51 and HPV45. Double infections occurred with the following genotypes: 52-53, 51-59, 61-67, 66-11, 16-62, 53-62, 59-CP6108, 45-66, and 45-51. Triple infections were identified as: 6-31-39, 51-59-62, 51-62-81, 54-55-59, 16-58-71, and 16-59-62. The prevalent genotype found among women from Michoacán, HPV59, was different to the rest of the country. The high prevalence of HPV59 could be due to cases imported to Michoacán by agricultural workers migrating to the USA or may be associated to ethnicity differences. Implications of this finding for immunization programs should be explored.

Seroprevalence of hepatitis B and delta viruses among HIV-infected population attending anti-retroviral clinic in selected health facilities in Abuja, Nigeria

ABSTRACTIntroduction: Triple infection with human immunodeficiency virus (HIV), hepatitis B virus (HBV), and hepatitis D virus (HDV) is rare. There is limited data on the seroprevalence of HIV/HBV/HDV tri-infection especially in Nigeria. The aim of this study was to determine the seroprevalences of HBsAg and HDV among HIV-infected individuals attending anti-retroviral (ARV) clinics in Abuja, Nigeria.Methods: In this cohort study, blood samples were collected from 1102 (male = 450; female = 652), with age range <20 to ≥51 years (mean age = 34.0; SD = 11.5), consenting HIV-infected population attending ARV clinics at selected health facilities in Abuja, Nigeria, between April and October 2016. A well-structured questionnaire was used to capture demographic information from the respondents. Enzyme-linked immunosorbent assay (ELISA) was used to determine the seroprevalence of hepatitis B surface antigen and anti-HDV. The result was interpreted according to manufacturer’s instruction. Statistical data were analyzed using SPSS software version 21, and chi-square (χ2) test was used to determine association with P < 0.05 considered significant.Results: Overall seroprevalences of 10.3%, 7.1%, and 0.7% for HBV, HBV/HDV, and HIV/HBV/HDV, respectively, were found among the study population. The infection rate (13.3%) peaked at age range of 31–40 years for HBV (P = 0.002), 50% at <20 years for HBV/HDV (P = 0.049), and 1.5% at 31–40 years for HIV/HBV/HDV (P = 0.202). By gender, the rate was higher in males (10.9%, 10.2%, 1.1%) than females (9.8%, 4.9%, 0.5%) for HBV, HBV/HDV, and HIV/HBV/HDV infections, respectively. However, there was no significant association between infection rate and gender.Conclusion: This study has established that HBV and HDV prevalence is still high in the population studied and that the rate of triple infection is low. We advocate for more robust control measures for HBV which should be extended to HDV in HIV population through screening and vaccination.

Occurrence and public health implications of gastrointestinal parasites of domesticated pigs (Sus scrofa domesticus) in Billiri Local Government Area, Gombe State, Nigeria

A cross-sectional study was designed, from February, 2016 to October, 2016 using systematic random sampling technique. For the determination of (GIT) parasites, faecal samples were collected from the study animals and subjected to faecal floatation and sedimentation technique. A.suum has the highest prevalence of 147 (24.5%), then followed by Trichuris suis 96 (16.0%), Emeria species 73 (12.2%), Oesophagostomum species 58 (9.7%), strongyloides spp 41 (6.8%), Fasciola species 28 (4.7%), Isospora suis 17 (2.8%), paragonimus suis 11 (1.8%), schistosoma suis 8 (1.3%) and Taenia solum 28 (0.3%) respectively. Pigs with single infection are (53.2%), double infections (18.5%) and triple infections (80.2%). For sex boars have prevalence of (34.0%) and sows have (46.2%) with P-value of 0.0007 which is statistically significant and relative risk of 1.208. For age the young have prevalence of (27.2%) and the adults (53.0%) with P-value of 0.0066 which is statistically significant and relative risk of 1.166. For management system the extensive have a prevalence of (64.7%) while the semi intensive (15.5%) with p-value of 0.0001 which is statistically significant and relative risk of 0.7884. Dry season has prevalence of (30.2%) and (50.0%) for rainy season with p-value of 0.0001 which is statistically significant and relative risk of 0.8017. The overall prevalence for households is (66.7%) out of 485 sample pigs while for abattoir is (13.5%) out of 115 sample pigs. The overall prevalence from this study is (80.2%) out of 600 pigs sampled.

'Candidatus Phytoplasma wodyetiae', a new taxon associated with yellow decline disease of foxtail palm (Wodyetia bifurcata) in Malaysia.

Landscape-grown foxtail palm (Wodyetia bifurcata A. K. Irvine) trees displaying symptoms of severe foliar chlorosis, stunting, general decline and mortality reminiscent of coconut yellow decline disease were observed in Bangi, Malaysia, during 2012. DNA samples from foliage tissues of 15 symptomatic palms were analysed by employing a nested PCR assay primed by phytoplasma universal ribosomal RNA operon primer pairs, P1/P7 followed by R16F2n/R2. The assay yielded amplicons of a single band of 1.25 kb from DNA samples of 11 symptomatic palms. Results from cloning and sequence analysis of the PCR-amplified 16S rRNA gene segments revealed that, in three palms, three mutually distinct phytoplasmas comprising strains related to 'Candidatus Phytoplasma asteris' and 'Candidatus Phytoplasma cynodontis', as well as a novel phytoplasma, were present as triple infections. The 16S rRNA gene sequence derived from the novel phytoplasma shared less than 96 % nucleotide sequence identity with that of each previously describedspecies of the provisional genus 'Ca. Phytoplasma', justifying its recognition as the reference strain of a new taxon, 'Candidatus Phytoplasma wodyetiae'. Virtual RFLP profiles of the R16F2n/R2 portion of the 16S rRNA gene and the pattern similarity coefficient value (0.74) supported the delineation of 'Ca. Phytoplasma wodyetiae' as the sole representative subgroup A member of a new phytoplasma ribosomal group, 16SrXXXVI.

Enteric viruses in HIV-1 seropositive and HIV-1 seronegative children with diarrheal diseases in Brazil.

Diarrheal diseases (DD) have distinct etiological profiles in immune-deficient and immune-competent patients. This study compares detection rates, genotype distribution and viral loads of different enteric viral agents in HIV-1 seropositive (n = 200) and HIV-1 seronegative (n = 125) children hospitalized with DD in Rio de Janeiro, Brazil. Except for group A rotavirus (RVA), which were detected through enzyme immunoassay, the other enteric viruses (norovirus [NoV], astrovirus [HAstV], adenovirus [HAdV] and bocavirus [HBoV]) were detected through PCR or RT-PCR. A quantitative PCR was performed for RVA, NoV, HAstV, HAdV and HBoV. Infections with NoV (19% vs. 9.6%; p<0.001), HBoV (14% vs. 7.2%; p = 0.042) and HAdV (30.5% vs. 14.4%; p<0.001) were significantly more frequent among HIV-1 seropositive children. RVA was significantly less frequent among HIV-1 seropositive patients (6.5% vs. 20%; p<0.001). Similarly, frequency of infection with HAstV was lower among HIV-1 seropositive children (5.5% vs. 12.8%; p = 0.018). Among HIV-1 seropositive children 33 (16.5%) had co-infections, including three enteric viruses, such as NoV, HBoV and HAdV (n = 2) and NoV, HAstV and HAdV (n = 2). The frequency of infection with more than one virus was 17 (13.6%) in the HIV-1 negative group, triple infection (NoV + HAstV + HBoV) being observed in only one patient. The median viral load of HAstV in feces was significantly higher among HIV-1 positive children compared to HIV-1 negative children. Concerning children infected with RVA, NoV, HBoV and HAdV, no statistically significant differences were observed in the medians of viral loads in feces, comparing HIV-1 seropositive and HIV-1 seronegative children. Similar detection rates were observed for RVA, HAstV and HAdV, whilst NoV and HBoV were significantly more prevalent among children with CD4+ T lymphocyte count below 200 cells/mm3. Enteric viruses should be considered an important cause of DD in HIV-1 seropositive children, along with pathogens more classically associated with intestinal infections in immunocompromised hosts.

Potential-Resolved Electrochemiluminescence for Simultaneous Determination of Triple Latent Tuberculosis Infection Markers.

A novel electrochemiluminescence (ECL) immunosensor based on the potential-resolved strategy was first developed for simultaneous determination of triple latent tuberculosis infection (LTBI) markers with high sensitivity, interferon-gamma (IFN-γ), tumor necrosis factor-alpha (TNF-α), and interleukin (IL)-2. In this work, luminol and self-prepared carbon quantum dots and CdS quantum dots were integrated onto gold nanoparticles and magnetic beads in sequence to fabricate potential-resolved ECL nanoprobes with signal amplification. IFN-γ-antibody (Ab)1, TNF-α-Ab1, and IL-2-Ab1 were separately immobilized on three spatially resolved areas of a patterned indium tin oxide electrode to capture the corresponding LTBI markers, which were further recognized by IFN-γ-Ab2, TNF-α-Ab2, and IL-2-Ab2-functionalized ECL nanoprobes. The binding reaction of antibody-functionalized ECL nanoprobes and the captured LTBI markers will generate three sensitive and potential-resolved ECL signals during one potential scanning, and the ECL intensities reflect the concentrations of IFN-γ, TNF-α, and IL-2 in the range of 1.6-200 pg mL-1. Therefore, the multiplexed ECL immunosensor provided an effective approach for simultaneous detection of triple LTBI markers in human serum, so it will be beneficial to facilitate more accurate and reliable clinical diagnosis for LTBI.

Viral Surveillance in Serum Samples From Patients With Acute Liver Failure By Metagenomic Next-Generation Sequencing.

Twelve percent of all acute liver failure (ALF) cases are of unknown origin, often termed indeterminate. A previously unrecognized hepatotropic virus has been suspected as a potential etiologic agent. We compared the performance of metagenomic next-generation sequencing (mNGS) with confirmatory nucleic acid testing (NAT) to routine clinical diagnostic testing in detection of known or novel viruses associated with ALF. Serum samples from 204 adult ALF patients collected from 1998 to 2010 as part of a nationwide registry were analyzed. One hundred eighty-seven patients (92%) were classified as indeterminate, while the remaining 17 patients (8%) served as controls, with infections by either hepatitis A virus or hepatitis B virus (HBV), or a noninfectious cause for their ALF. Eight cases of infection from previously unrecognized viral pathogens were detected by mNGS (4 cases of herpes simplex virus type 1, including 1 case of coinfection with HBV, and 1 case each of HBV, parvovirus B19, cytomegalovirus, and human herpesvirus 7). Several missed dual or triple infections were also identified, and assembled viral genomes provided additional information on genotyping and drug resistance mutations. Importantly, no sequences corresponding to novel viruses were detected. These results suggest that ALF patients should be screened for the presence of uncommon viruses and coinfections, and that most cases of indeterminate ALF in the United States do not appear to be caused by novel viral pathogens. In the future, mNGS testing may be useful for comprehensive diagnosis of viruses associated with ALF, or to exclude infectious etiologies.

Infections with Schistosoma mansoni and geohelminths among school children dwelling along the shore of the Lake Hawassa, southern Ethiopia.

BackgroundInfections with Schistosoma mansoni and soil-transmitted helminthes (STHs) are major public health problems in Ethiopia. However, information was scarce on the current status of these infections to guide an intervention in the study area. Therefore, this study was conducted to assess the prevalence of infections with S. mansoni and STHs and associated factors among school children in southern Ethiopia.MethodsThis cross-sectional study investigated children who were attending the Finchawa and Tullo junior elementary schools and were residing along the shore of the lake Hawassa in January and February, 2015. A total of 374 students were selected using systematic random sampling technique. Data on socio-demography and related factors was collected using structured questionnaires. A single stool sample was collected from each child and processed using formol-ether concentration technique and examined microscopically for parasites’ ova/larva.ResultsThe prevalence of parasitic infection with one or more than one helminthiasis was found to be 67.9%. Seven different types of helminths were identified and the most prevalent parasites were Ascaris lumbricoides (44.4%), followed by S. mansoni (31%), Trichuris trichiura (11%), and hookworms (7.7%). The rate of infection with STHs was 52.4%. Single, double, triple and quadruple infections were 42.2, 22.5, 2.4, and 0.8%, respectively. Children who practiced open-field defecation (AOR, 3.6; 95% CI 1.6–8.0; p = 0.001) and had not always washed their hands before eating a meal (AOR, 5.0; 95%CI 2.15–11.7; p <0.001) were more infected with STHs. Moreover, the rate of S. mansoni infection was significantly higher among children who were attending the Finchawa school (AOR, 2.13; 95% CI 1.31–3.46; p = 0.002), aged 11–15 years (AOR, 1.97; 95% CI 1.22–3.19; p = 0.006), had swum in the lake Hawassa (AOR, 2.73; 95%CI 1.20–6.17; p = 0.016), and had involved in irrigation-related activities (AOR, 1.68; 95%CI 1.04–2.71; p = 0.034).ConclusionThe study showed high prevalence of STHs and moderate rate of schisotosomiasis. Mass deworming twice a year for STHs and once every two years for S. mansoni, further to strengthening other prevention measures, is critically needed to reduce these infections to inconsequential level.

P212 The clap trap: the effectiveness of partner notification for gonorrhoea in MSM in an lgbt community sexual health clinic

IntroductionHigh rates of partner change, multiple casual partners, and complex sexual networks are thought to contribute to outbreaks of Neisseria gonorrhoea (GC) in men who have sex with men (MSM), who account for 42% of diagnoses in the UK. Prompt, appropriate treatment and effective partner notification are key to reducing transmission. We audited partner notification for GC in MSM attending an LGBT community sexual health clinic.MethodsA retrospective audit over 12 months (2015–16). A total of 33 episodes of GC were diagnosed in 31 patients. Data was recorded on a spreadsheet for analysis.Results25(76%) were asymptomatic. 29(88%) underwent triple site testing and 4(12%) dual site. Gonorrhoea was detected in the pharynx in 23(70%), urethra in 7(21%), and rectum in 14(42%). 7(21%) had dual and 2(6%) triple site infection. 29/33(88)% were informed of the diagnosis within 10 days (target 95%). 28/33(85%) were treated within 2 weeks. 6 attended as contacts of GC and were treated on the day they attended. A total of 109 contacts were given. 50(46%) were untraceable. Of the traceable contacts, 31/59(53%) were confirmed as treated, 23 at the same clinic. A total of 0.9 contacts were treated per index case (target 0.6).DiscussionThe high frequency of unknown casual partners in MSM with GC means there is often inadequate information to trace partners. Nevertheless, this audit has shown good outcomes for partner notification in a community LGBT sexual health clinic. This should contribute to reducing onward transmission in a high risk MSM population.

Impact of simultaneous exposure to arboviruses on infection and transmission by Aedes aegypti mosquitoes

The recent emergence of both chikungunya and Zika viruses in the Americas has significantly expanded their distribution and has thus increased the possibility that individuals may become infected by more than one Aedes aegypti-borne virus at a time. Recent clinical data support an increase in the frequency of coinfection in human patients, raising the likelihood that mosquitoes could be exposed to multiple arboviruses during one feeding episode. The impact of coinfection on the ability of relevant vector species to transmit any of these viruses (that is, their vector competence) has not been determined. Thus, we here expose Ae. aegypti mosquitoes to chikungunya, dengue-2 or Zika viruses, both individually and as double and triple infections. Our results show that these mosquitoes can be infected with and can transmit all combinations of these viruses simultaneously. Importantly, infection, dissemination and transmission rates in mosquitoes are only mildly affected by coinfection.

Spread of poxviruses in livestock in Brazil associated with cases of double and triple infection.

The objective of this work is to describe the distribution of outbreaks of vaccinia virus (VACV), pseudocowpox virus (PCPV), and bovine papular stomatitis virus (BSPV) in Brazil. The Official Laboratory of the Brazilian Ministry of Agriculture received 89 samples from different locations in Brazil in 2015 and 2016 for diagnosis of vesicular and exanthematous disease. Poxvirus coinfections occurred in 11 out of 33 outbreaks, including the first reported triple infection by BPSV, PCPV, and VACV. This occurrence may be associated with the circulation of these viruses in Brazilian cattle.

Delineating morbillivirus entry, dissemination and airborne transmission by studying in vivo competition of multicolor canine distemper viruses in ferrets.

Identification of cellular receptors and characterization of viral tropism in animal models have vastly improved our understanding of morbillivirus pathogenesis. However, specific aspects of viral entry, dissemination and transmission remain difficult to recapitulate in animal models. Here, we used three virologically identical but phenotypically distinct recombinant (r) canine distemper viruses (CDV) expressing different fluorescent reporter proteins for in vivo competition and airborne transmission studies in ferrets (Mustela putorius furo). Six donor ferrets simultaneously received three rCDVs expressing green, red or blue fluorescent proteins via conjunctival (ocular, Oc), intra-nasal (IN) or intra-tracheal (IT) inoculation. Two days post-inoculation sentinel ferrets were placed in physically separated adjacent cages to assess airborne transmission. All donor ferrets developed lymphopenia, fever and lethargy, showed progressively increasing systemic viral loads and were euthanized 14 to 16 days post-inoculation. Systemic replication of virus inoculated via the Oc, IN and IT routes was detected in 2/6, 5/6 and 6/6 ferrets, respectively. In five donor ferrets the IT delivered virus dominated, although replication of two or three different viruses was detected in 5/6 animals. Single lymphocytes expressing multiple fluorescent proteins were abundant in peripheral blood and lymphoid tissues, demonstrating the occurrence of double and triple virus infections. Transmission occurred efficiently and all recipient ferrets showed evidence of infection between 18 and 22 days post-inoculation of the donor ferrets. In all cases, airborne transmission resulted in replication of a single-colored virus, which was the dominant virus in the donor ferret. This study demonstrates that morbilliviruses can use multiple entry routes in parallel, and co-infection of cells during viral dissemination in the host is common. Airborne transmission was efficient, although transmission of viruses expressing a single color suggested a bottleneck event. The identity of the transmitted virus was not determined by the site of inoculation but by the viral dominance during dissemination.

Hepatitis B and C Virus Infections Among Human Immunodeficiency Virus-Infected People Who Inject Drugs in Lahore, Pakistan.

Hepatitis B virus (HBV) and hepatitis C virus (HCV) are the major cause of the global burden of hepatitis. One of the main routes of transmission for both viruses is through exposure to infected blood, which includes sharing blood-contaminated syringes and needles. Human immunodeficiency virus (HIV) attacks the immune system and results in acquired immune deficiency syndrome and opportunistic infections. The objective of this study was to assess the epidemiology of HBV and HCV infections among HIV-infected people who inject drugs (PWID). The study enrolled 100 PWID from different addiction centers of the city of Lahore in Pakistan. All subjects were HIV-infected males and were above 16 years of age. Screening of HBV and HCV infections was performed through immunochromatography tests and enzyme-linked immunosorbent assays. The prevalence of HCV and HBV infections among the 100 HIV-infected PWID was 55% and 6%, respectively. HIV monoinfection was found in 37% of the subjects, while triple infection was detected in 2% of the subjects. Majority of the HIV-infected PWID were using heroin and Avil injections (65%). Half of the subjects had used injection drugs for 1-5 years, while 32% had used injection drugs for 6-10 years. HCV infection was more common than HBV infection among the enrolled subjects. Most of the PWID were practicing heroin and Avil injections.

One-step multiplex RT-qPCR detects three citrus viroids from different genera in a wide range of hosts

A one-step multiplex reverse transcription real-time quantitative polymerase chain reaction (RT-qPCR) based on species-specific minor groove binding (MGB) probes, was developed for the simultaneous detection, identification, and quantification of three citrus viroids belonging to different genera. Citrus exocortis viroid (Pospiviroid), Hop stunt viroid (Hostuviroid), and Citrus bark cracking viroid (Cocadviroid) cause a variety of maladies in agriculturally significant crops. Therefore, reliable assays for their detection are essential tools for various government and industry organizations implementing disease management programs. Singleplex qPCR primers and MGB probes were designed individually for the detection of the three targeted viroids, and subsequently combined in a one-step multiplex RT-qPCR reaction. A wide host range of woody plants, including citrus, grapevines, apricots, plums and herbaceous plants such as tomato, cucumber, eggplant and chrysanthemum different world regions were used to validate the assay. Single, double and triple viroid infections were identified in the tested samples. The developed multiplex RT-qPCR assay was compared with a previously reported SYBR Green I RT-qPCR for the universal detection of citrus viroids. Both assays accurately identified all citrus viroid infected samples. The multiplex assay complemented the SYBR Green I universal detection assay by differentiating among citrus viroid species in the positive samples. The developed multiplex RT-qPCR assay has the potential to simultaneously detect each targeted viroid and could be used in high throughput screenings for citrus viroids in field surveys, germplasm banks, nurseries and other viroid disease management programs.