Surveillance of leishmaniasis cases from 15 European centres, 2014 to 2019: a retrospective analysis.

BackgroundSurveillance of human leishmaniasis in Europe is mostly limited to country-specific information from autochthonous infections in the southern part. As at the end of 2021, no integrated analysis has been performed for cases seen across centres in different European countries.AimTo provide a broad perspective on autochthonous and imported leishmaniasis cases in endemic and non-endemic countries in Europe.MethodsWe retrospectively collected records from cutaneous, mucosal and visceral leishmaniasis cases diagnosed in 15 centres between 2014 and 2019. Centres were located in 11 countries: Belgium, France, Germany, Italy, the Netherlands, Norway, Portugal, Spain, Sweden, Switzerland and the United Kingdom. Data on country of infection, reason for travelling, infecting species, age and sex were analysed.ResultsWe obtained diagnostic files from 1,142 cases, of which 76%, 21% and 3% had cutaneous, visceral, and mucosal disease, respectively. Of these, 68% were men, and 32% women, with the median age of 37 years (range: 0-90) at diagnosis. Visceral leishmaniasis was mainly acquired in Europe (88%; 167/190), while cutaneous leishmaniasis was primarily imported from outside Europe (77%; 575/749). Sixty-two percent of cutaneous leishmaniasis cases from outside Europe were from the Old World, and 38% from the New World. Geographic species distribution largely confirmed known epidemiology, with notable exceptions.ConclusionsOur study confirms previous reports regarding geographic origin, species, and traveller subgroups importing leishmaniasis into Europe. We demonstrate the importance of pooling species typing data from many centres, even from areas where the aetiology is presumably known, to monitor changing epidemiology.

Sexual Risk Behavior in HIV-Uninfected Pregnant Women in Western Uganda.

Our aim was to identify sexual risk behavior among HIV-negative pregnant women in Kabarole District, Uganda, by conducting a cross-sectional study among 1610 women within three healthcare settings. One in six women engaged in HIV-specific risk behaviors including multiple sexual partners or alcohol abuse; 80% of the pregnant women reported to generally abstain from using condoms. In multivariate analysis, predictors of sexual risk behavior included being a client of the public health facilities as compared to the private facility (AOR 3.6 and 4.8, p < 0.001), being single, widowed or divorced or not cohabiting with the partner (AOR 4.7 and 2.3, p < 0.001), as well as higher household wealth (AOR 1.8, p < 0.001) and lack of partner status knowledge (AOR 1.6, p = 0.008). Self-estimated risk perception was linked with engagement in HIV-related risk behaviors except for alcohol abuse. Our findings indicate that reducing risky behaviors in pregnancy in order to prevent HIV should be a high-priority public health concern.

"In the hospital, there will be nobody to pamper me": a qualitative assessment on barriers to facility-based delivery in post-Ebola Sierra Leone.

BACKGROUND: Sierra Leone has one of the highest maternal mortality rates in the world. Encouraging the use of skilled birth attendance in health facilities is an important step in the endeavor to increase the number of safe deliveries. However, public trust in health facilities has been greatly damaged during the Ebola epidemic outbreak in Sierra Leone in 2014/2015, and little is known about external and intrinsic barriers to facility-based delivery (FBD) in the country since the end of the Ebola epidemic. METHODS: We conducted a qualitative study on FBD in Princess Christian Maternity Hospital, Freetown, which is the national referral maternity hospital in Sierra Leone. We performed six focus group discussions with providers, pregnant women and recent mothers surrounding experiences, attitudes and behaviors regarding FBD and potential barriers. Discussions were tape recorded, transcribed and evaluated through content analysis. RESULTS: Women in our study were overall technically aware of the higher safety linked with FBD, but this often diverged from their individual desire to deliver in a supportive and trusted social and traditional environment. Close relatives and community members seemed to be highly influencial regarding birth practices. Many women associated FBD with negative staff attitudes and an undefined fear. Logistic issues regarding transportation problems or late referral from smaller health centers were identified as frequent barriers to FBD. CONCLUSIONS: More supportive staff attitudes and acceptance of an accompanying person throughout delivery could be promising approaches to increase women's confidence in FBDs. However, these approaches also imply revising health systems structures, like staff working conditions that are conducive for a friendly atmosphere, sufficient space in delivery wards allowing the women to bring a birth companion, or like the establishment of a reliable peripheral ambulance system to ensure transportation and fast referral.

Option B+ for prevention of vertical HIV transmission has no influence on adverse birth outcomes in a cross-sectional cohort in Western Uganda.

BACKGROUND: While most Sub-Saharan African countries are now implementing the WHO-recommended Option B+ protocol for prevention of vertical HIV transmission, there is a lack of knowledge regarding the influence of Option B+ exposure on adverse birth outcomes (ABOs). Against this background, we assessed ABOs among delivering women in Western Uganda. METHODS: A cross-sectional, observational study was performed within a cohort of 412 mother-newborn-pairs in Virika Hospital, Fort Portal in 2013. The occurrence of stillbirth, pre-term delivery, and small size for gestational age (SGA) was analysed, looking for influencing factors related to HIV-status, antiretroviral drug exposure and duration, and other sociodemographic and clinical parameters. RESULTS: Among 302 HIV-negative and 110 HIV-positive women, ABOs occurred in 40.5%, with stillbirth in 6.3%, pre-term delivery in 28.6%, and SGA in 12.2% of deliveries. For Option B+ intake (n = 59), no significant association was found with stillbirth (OR 0.48, p = 0.55), pre-term delivery (OR 0.97, p = 0.92) and SGA (OR 1.5, p = 0.3) compared to seronegative women. Women enrolled on antiretroviral therapy (ART) before conception (n = 38) had no different risk for ABOs than women on Option B+ or HIV-negative women. Identified risk factors for stillbirth included lack of formal education, poor socio-economic status, long travel distance, hypertension and anaemia. Pre-term delivery risk was increased with poor socio-economic status, primiparity, Malaria and anaemia. The occurrence of SGA was influenced by older age and Malaria. CONCLUSION: In our study, women on Option B+ showed no difference in ABOs compared to HIV-negative women and to women on ART. We identified several non-HIV/ART-related influencing factors, suggesting an urgent need for improving early risk assessment mechanisms in antenatal care through better screening and triage systems. Our results are encouraging with regard to continued universal scale-up of Option B+ and ART programmes.

Screening for infectious diseases among unaccompanied minor refugees in Berlin, 2014-2015.

Infectious diseases (except tuberculosis) were screened among 1248 unaccompanied minor refugees (UMRs) arriving in Berlin in 2014-2015; 40 % originated from Syria. More than half of the refugees presented without any pathologic finding. Infections requiring treatment were diagnosed in 19.6 %, mainly infections with Giardia and intestinal helminths as well as schistosomiasis, while potentially contagious diseases were diagnosed in 15.3 % of all screened UMRs.

Comparison of Leishmania typing results obtained from 16 European clinical laboratories in 2014.

Leishmaniasis is endemic in southern Europe, and in other European countries cases are diagnosed in travellers who have visited affected areas both within the continent and beyond. Prompt and accurate diagnosis poses a challenge in clinical practice in Europe. Different methods exist for identification of the infecting Leishmania species. Sixteen clinical laboratories in 10 European countries, plus Israel and Turkey, conducted a study to assess their genotyping performance. DNA from 21 promastigote cultures of 13 species was analysed blindly by the routinely used typing method. Five different molecular targets were used, which were analysed with PCR-based methods. Different levels of identification were achieved, and either the Leishmania subgenus, species complex, or actual species were reported. The overall error rate of strains placed in the wrong complex or species was 8.5%. Various reasons for incorrect typing were identified. The study shows there is considerable room for improvement and standardisation of Leishmania typing. The use of well validated standard operating procedures is recommended, covering testing, interpretation, and reporting guidelines. Application of the internal transcribed spacer 1 of the rDNA array should be restricted to Old World samples, while the heat-shock protein 70 gene and the mini-exon can be applied globally.

Lack of effect of intermittent preventive treatment for malaria in pregnancy and intense drug resistance in western Uganda.

BACKGROUND: Intermittent preventive treatment in pregnancy (IPTp) with sulfadoxine-pyrimethamine (SP) is widely implemented in sub-Saharan Africa for the prevention of malaria in pregnancy and adverse birth outcomes. However, in areas of intense SP resistance, the efficacy of IPTp may be compromised. METHODS: A cross-sectional study among 915 delivering women (728 analysable live singleton deliveries) was conducted in Fort Portal, western Uganda, to assess associations of reported IPTp use, Plasmodium falciparum infection, maternal anaemia, low birth weight, and preterm delivery, and to estimate the degree of SP resistance as reflected by pfdhfr/pfdhps mutations. RESULTS: Plasmodium falciparum infection was detected by PCR in 8.9 % and by microscopy of placental blood samples in 4.0 %. Infection was significantly associated with stillbirth, early neonatal death, anaemia, low birth weight, and pre-term delivery. Eighty percent of the women had taken at least one dose of IPTp, and more than half had taken two doses. As compared to women without chemoprophylaxis against malaria, IPTp had no significant influence on the presence of P. falciparum infection (13.8 vs. 9.6 %, P = 0.31). Nor was it associated with reductions in anaemia, low birth weight or preterm delivery. P. falciparum with intense SP resistance (pfdhfr/pfdhps quintuple or sextuple mutations) were observed in 93 % (pfdhps 581G, 36 %), and the additional high resistance allele pfhdr 164L in 36 %. CONCLUSIONS: In Fort Portal, Uganda, reported use of IPTp with SP does not provide an observable benefit. The molecular markers of P. falciparum indicate high grade SP resistance reaching the threshold set by WHO for the discontinuation of IPTp with SP. Alternative approaches for the prevention of malaria in pregnancy are urgently needed.

Soil-transmitted helminths in southern highland Rwanda: associated factors and effectiveness of school-based preventive chemotherapy.

OBJECTIVES: Preventive chemotherapy of schoolchildren against soil-transmitted helminths (STHs) is widely implemented in Rwanda. However, data on its actual efficacy are lacking. We assessed prevalence, associated factors and manifestation of STH infection among schoolchildren in southern highland Rwanda as well as cure and reinfection rates. METHODS: Six hundred and twenty-two children (rural, 301; urban, 321) were included preceding the administration of a single dose of 500 mg mebendazole. Before treatment, and after 2 and 15 weeks, STH infection was determined by Kato-Katz smears and by PCR assays for Ascaris lumbricoides. Clinical and anthropometric data, socio-economic status and factors potentially associated with STH infection were assessed. RESULTS: Soil-transmitted helminth (STH) infection was present in 38% of rural and in 13% of urban schoolchildren. Ascaris lumbricoides accounted for 96% of infections. Of these, one-third was detected by PCR exclusively. Factors associated with STH infection differed greatly between rural and urban children. Likewise, STH infection was associated with stunting and anaemia only among urban children. The cure rate after 2 weeks was 92%. Among eight non-cleared A. lumbricoides infections, seven were submicroscopic. Reinfection within 3 months occurred in 7%, but the rate was higher among rural children, and with initially present infection, particularly at comparatively high intensity. CONCLUSIONS: The rural-urban difference in factors associated with STH infection and in reinfection rates highlights the need for targeted interventions to reduce transmission. PCR assays may help in detecting low-level infections persisting after treatment. In southern Rwanda, mebendazole is highly effective against the STH infections predominated by A. lumbricoides.

The challenge of referring HIV-positive pregnant women with treatment indication from PMTCT to ART services: a retrospective follow-up study in Mbeya, Tanzania.

Providing full antiretroviral therapy (ART) to all HIV-positive, pregnant women with treatment indication could significantly reduce overall mother-to-child transmission. However, the effectiveness of referring HIV-positive antenatal care (ANC) clients with a treatment indication to ART services has rarely been assessed to date. We retrospectively followed-up data of a cohort of treatment-eligible ANC clients in Mbeya Region, Tanzania by retracing and merging registries of ANC, Care and Treatment Centers (CTC), and Infant Care. ART initiation and ART duration before delivery served as primary outcome indicators to assess referral effectiveness. We retraced data of 60 ANC clients with treatment indication: 39 (65%) started predelivery ART and 21 (35%) remained untreated during pregnancy. Eight (13.3%) did not initiate ART at all within the observation period. Women starting ART before delivery had significantly lower CD4-cell counts at enrollment than nonstarters (medians: 207.5 vs. 292 cells/µl; p = 0.013). Predelivery ART starters had experienced a significantly shorter duration between staff-declared "ART readiness" and actual ART start (medians: 0 vs. 28 days; p = 0.0004). The median ART duration prior to delivery was 57 days; only eight women (13.3%) accomplished ≥90 days ART intake during pregnancy. Early enrollment in ANC at ≤24 gestational weeks was associated with longer duration of predelivery ART. At maternity wards, 24.3% of treatment-eligible mothers and newborns with retraceable delivery data had received no or inadequate antiretrovirals. Within 6 months postdelivery, women attended on average 3.5 out of 6 requested CTC visits. Concluding, every third treatment-eligible woman in this cohort was not covered through ART before delivery, and predelivery ART duration was mostly suboptimal regarding vertical transmission prevention. HIV-positive women need to be encouraged to approach ANC early in pregnancy, and health services need to address unnecessary time gaps before ART initiation. In addition, inclusive ART services for HIV-positive ANC clients should be seriously discussed.

American Cutaneous Leishmaniasis: Imported cases in Berlin 2000-2023.

BACKGROUND: American Cutaneous Leishmaniasis (ACL) shows variable response to therapy, but data on species-specific treatment efficacy is scarce. We describe the clinical characteristics and outcome of patients with ACL imported to a tertiary centre in Germany and determine whether species-specific therapy according to the 2014 "LeishMan" group recommendations is associated with cure. METHODS: A retrospective chart review was conducted at the Charité Institute of International Health in Berlin. We analysed data on PCR-confirmed ACL cases collected between 2000 and 2023. Systemic therapy included liposomal amphotericin B, miltefosine, pentavalent antimony, ketoconazole or itraconazole. Localized therapy included perilesional pentavalent antimony or paromomycin ointment. Cure was defined as re-epithelialization of ulcers or disappearance of papular-nodular lesions after 3 months of treatment. Logistic regression models were used to quantify the effect of species-specific systemic therapy on the outcome. RESULTS: 75 cases were analysed. Most patients were male (62%), median age was 35 years, no patient had a history of immunosuppression. The most common reason for travel was tourism (60%), the most common destination was Costa Rica (28%), the median duration of illness was 8 weeks, and most patients presented with ulcers (87%). Lesions were complex in 43%. The most common Leishmania (L.) species was L. braziliensis (28%), followed by L. panamensis (21%). 51/73 (70%) patients were cured after initial therapy and 17/21 (81%) after secondary therapy. Cure after systemic therapy was more frequent when species-specific treatment recommendations were followed (33/45; 73%), compared to when not followed, (6/17; 35%, P = 0.008). This association was independent of age, sex, previous therapy, complex lesions, and Leishmania species (adjusted OR, 5.06; 95% CI, 1.22-24.16). CONCLUSIONS: ACL is a rare, imported disease in Germany. Complex lesions were common, challenging successful therapy. This study highlights the importance of identifying the parasite species and suggests that a species-specific approach to treatment leads to better outcomes.

Treatment outcome of imported cutaneous leishmaniasis among travelers and migrants infected with Leishmania major and Leishmania tropica: a retrospective study in European centers 2013 to 2019.

OBJECTIVES: Cutaneous leishmaniasis (CL) in Asia, Northern, and Sub-Saharan Africa is mainly caused by Leishmania major and Leishmania tropica. We describe and evaluate the treatment outcome of CL among travelers and migrants in Europe. METHODS: We conducted a retrospective study of parasitological confirmed CL cases caused by L. major and L. tropica during 2013-2019 in Europe. Data were collected from medical records and databases within the LeishMan network. RESULTS: Of 206 included cases of CL, 75 were identified as L. major and 131 as L. tropica. Of patients with L. tropica infection, 80% were migrants, whereas 53% of patients with L. major infection had been visiting friends and relatives. Among patients with L. tropica, 48% were younger than 15 years. Pentavalent antimony cured 73% (L. major) and 78% (L. tropica) of patients. The cure rate for intralesional administration was 86% and 67% for systemic, on L. tropica. Liposomal amphotericin B had a cure rate of 44-63%. CONCLUSION: L. major infections were mostly found in individuals visiting friends and relatives, whereas L. tropica were mainly identified in migrants. No patients with L. major relapsed. Pentavalent antimony, liposomal amphotericin B, and cryotherapy had cure rates in accordance with previous studies.

Prevalence and risk factors of malaria among children in southern highland Rwanda.

BACKGROUND: Increased control has produced remarkable reductions of malaria in some parts of sub-Saharan Africa, including Rwanda. In the southern highlands, near the district capital of Butare (altitude, 1,768 m), a combined community-and facility-based survey on Plasmodium infection was conducted early in 2010. METHODS: A total of 749 children below five years of age were examined including 545 randomly selected from 24 villages, 103 attending the health centre in charge, and 101 at the referral district hospital. Clinical, parasitological, haematological, and socio-economic data were collected. RESULTS: Plasmodium falciparum infection (mean multiplicity, 2.08) was identified by microscopy and PCR in 11.7% and 16.7%, respectively; 5.5% of the children had malaria. PCR-based P. falciparum prevalence ranged between 0 and 38.5% in the villages, and was 21.4% in the health centre, and 14.9% in the hospital. Independent predictors of infection included increasing age, low mid-upper arm circumference, absence of several household assets, reported recent intake of artemether-lumefantrine, and chloroquine in plasma, measured by ELISA. Self-reported bed net use (58%) reduced infection only in univariate analysis. In the communities, most infections were seemingly asymptomatic but anaemia was observed in 82% and 28% of children with and without parasitaemia, respectively, the effect increasing with parasite density, and significant also for submicroscopic infections. CONCLUSIONS: Plasmodium falciparum infection in the highlands surrounding Butare, Rwanda, is seen in one out of six children under five years of age. The abundance of seemingly asymptomatic infections in the community forms a reservoir for transmission in this epidemic-prone area. Risk factors suggestive of low socio-economic status and insufficient effectiveness of self-reported bed net use refer to areas of improvable intervention.

Mucosal Leishmania infantum infection.

Mucosal leishmaniasis is a well-known clinical manifestation of infections caused by species belonging to the Leishmania (Viannia) subgenus in Central and South America but not of Leishmania species endemic in the so-called Old World. We report on three cases of mucosal leishmaniasis caused by Leishmania (Leishmania) infantum contracted in southern Europe. Two patients were immunocompromised; one patient had no underlying condition. Lesions were located in the oral mucosa, oesophagus and nose. All lesions relapsed under standard treatment with liposomal amphotericin B. A cure was achieved after secondary and extended treatment with liposomal amphotericin B or miltefosine. Mucosal leishmaniasis contracted in southern Europe has to be considered in the differential diagnosis of lesions in the naso-buccal-oesophageal mucosa and may occur in previously healthy persons.

Clinical diversity and treatment results in Tegumentary Leishmaniasis: A European clinical report in 459 patients.

BACKGROUND: Cutaneous leishmaniasis (CL) is frequent in travellers and can involve oro-nasal mucosae. Clinical presentation impacts therapeutic management. METHODOLOGY: Demographic and clinical data from 459 travellers infected in 47 different countries were collected by members of the European LeishMan consortium. The infecting Leishmania species was identified in 198 patients. PRINCIPAL FINDINGS: Compared to Old World CL, New World CL was more frequently ulcerative (75% vs 47%), larger (3 vs 2cm), less frequently facial (17% vs 38%) and less frequently associated with mucosal involvement (2.7% vs 5.3%). Patients with mucosal lesions were older (58 vs 30 years) and more frequently immunocompromised (37% vs 3.5%) compared to patients with only skin lesions. Young adults infected in Latin America with L. braziliensis or L. guyanensis complex typically had an ulcer of the lower limbs with mucosal involvement in 5.8% of cases. Typically, infections with L. major and L. tropica acquired in Africa or the Middle East were not associated with mucosal lesions, while infections with L. infantum, acquired in Southern Europe resulted in slowly evolving facial lesions with mucosal involvement in 22% of cases. Local or systemic treatments were used in patients with different clinical presentations but resulted in similarly high cure rates (89% vs 86%). CONCLUSION/SIGNIFICANCE: CL acquired in L. infantum-endemic European and Mediterranean areas displays unexpected high rates of mucosal involvement comparable to those of CL acquired in Latin America, especially in immunocompromised patients. When used as per recommendations, local therapy is associated with high cure rates.

Partner involvement in perinatal care and PMTCT services in Mbeya Region, Tanzania: the providers' perspective.

Partner involvement is considered to increase the effectiveness of female-oriented services for sexual and reproductive health (SRH), like those for antenatal care (ANC) or the prevention of mother-to-child transmission of HIV (PMTCT). However, male participation rates remain mostly low, and previous research has identified restrictive provider attitudes among barriers for partner participation in such services. Individual perspectives and experiences of healthcare providers are assumed to significantly influence the quality of delivered services. This study aimed at exploring providers' attitudes regarding partner involvement in ANC/PMTCT and other SRH services. A hundred interviews based on a semi-structured questionnaire were conducted among healthcare providers employed in an ANC-based PMTCT program in Mbeya Region, Tanzania. Interviewees expressed overall approval of male partner integration into the services, but this approval decreased when specifying for different service types, especially in those related to perinatal examinations or labor and delivery. Divergence between general attitudes and self-reported individual behavior was observed, querying the reliability of expressed attitudes. Among providers having at least one child, personally experienced partner attendance and approval of partner involvement were significantly associated for most service types. Although general views on partner involvement in SRH services seem to be mostly supportive, there is a need for health services to strengthen providers' positions toward male involvement, for example by communicating clear policies and job guidelines, and by encouraging partner service attendance among providers themselves.

The incidence of HIV and associated risk factors among pregnant women in Kabarole District, Uganda.

OBJECTIVES: The study attempted to determine the incidence of HIV among pregnant women in Kabarole District, Uganda, and to identify socio-demographic and behavioral risk factors for seroconversion during pregnancy. METHODS: We carried out a retrospective cohort study among women for whom a documented HIV-negative test result from the first pregnancy trimester could be confirmed using available records, and who were HIV-retested in the third trimester or during delivery. In total, 1610 pregnant women from three different healthcare settings took part in the study. We captured the results of repeated HIV tests and conducted semi-structured interviews to explore participants' socio-demographic characteristics and sexual risk behavior. For HIV incidence rates, we calculated the number of seroconversions per 100 person-years. We used Fisher's exact test to test for potential associations. Penalized maximum likelihood logistic regression and Poisson regression were applied to adjust for potential confounders. RESULTS: The overall HIV incidence rate among participants was 2.9/100 women-years. Among socio-demographic characteristics, the multivariable analysis showed a significant association of marital status with HIV incidence in pregnancy (IRR 8.78, 95%CI [1.13-68.33]). Risky sexual behaviors including higher number of sexual partners in pregnancy (IRR 2.78 [1.30-5.94]), unprotected sex with unknown persons (IRR 14.25 [4.52-44.93]), alcohol abuse (IRR 12.08 [4.18-34.90]) and sex under the influence of drugs or alcohol (IRR 6.33 [1.36-29.49]) were significantly associated with seroconversion in pregnancy (similar results in logistic regression). CONCLUSIONS: HIV incidence was three times higher among our pregnant study population compared to the general female population in Uganda. This underlines the importance of HIV prevention and repeat testing during pregnancy. Identified risk groups should be considered for pre-exposure prophylaxis.

Cutaneous leishmaniasis in refugees from Syria: complex cases in Berlin 2015-2020.

BACKGROUND: The Syrian conflict has led to a dramatic increase of Old World cutaneous leishmaniasis (CL), triggered by continuous population displacements, disrupted control programmes, poor shelter and sanitation. METHODS: A retrospective patient record study was conducted at the Institute of Tropical Medicine and International Health in Berlin. Records of all refugees from Syria treated for CL between January 2015 and March 2020 were reviewed. RESULTS: Twenty refugees from Syria were treated. Seventeen refugees (85%) had complex lesions, mainly due to previous therapy failure or localization on the face. A long disease duration (50% > 1 year), pronounced facial scarring (20%), recurrences (20%), or worsening of existing lesions (20%) were observed. Nine patients (45%) had been pretreated in Syria. Complete remission was achieved in 10 of 16 patients (63%) treated with perilesional antimony. Eight patients (40%) required systemic treatment, thereof four (20%) repeated systemic treatment. Eight patients (40%) reported a delay of therapy ≥3 months in Germany, thereof one patient with a delay of 12 months and one patient with a delay of 32 months. CONCLUSION: Between 2015 and 2020, Syrian refugees presented with severe morbidities of CL frequently requiring systemic and even consecutive systemic treatments. We assume a combination of socioeconomic and environmental factors associated with the ongoing Syrian conflict and migration to be responsible for the complex clinical presentations in this case series. More attention should be drawn to the situation of Syrian refugees with CL in countries where they are displaced to.

Detection and quantification of minor human immunodeficiency virus type 1 variants harboring K103N and Y181C resistance mutations in subtype A and D isolates by allele-specific real-time PCR.

Nevirapine (single dose), commonly used to prevent the mother-to-child transmission of human immunodeficiency virus (HIV) in developing countries, frequently induces viral resistance. Even mutations which occur only in a minor population of the HIV quasispecies (<20%) are associated with subsequent treatment failure but cannot be detected by population-based sequencing. We developed sensitive allele-specific real-time PCR (ASPCR) assays for two key resistance mutations of nevirapine. The assays were specifically designed to analyze HIV-1 subtype A and D isolates accounting for the majority of HIV infections in Uganda. Assays were evaluated using DNA standards and clinical samples of Ugandan women having preventively taken single-dose nevirapine. Lower detection limits of drug-resistant HIV type 1 (HIV-1) variants carrying reverse transcriptase mutations were 0.019% (K103N [AAC]), 0.013% (K103N [AAT]), and 0.29% (Y181C [TGT]), respectively. Accuracy and precision were high, with coefficients of variation (the standard ratio divided by the mean) of 0.02 to 0.15 for intra-assay variability and those of 0.07 to 0.15 (K103N) and 0.28 to 0.52 (Y181C) for inter-assay variability. ASPCR assays enabled the additional identification of 12 (20%) minor drug-resistant HIV variants in the 20 clinical Ugandan samples (3 mutation analyses per patient; 60 analyses in total) which were not detectable by population-based sequencing. The individual patient cutoff derived from the clinical baseline sample was more appropriate than the standard-based cutoff from cloned DNA. The latter is a suitable alternative since the presence/absence of drug-resistant HIV-1 strains was concordantly identified in 92% (55/60) of the analyses. These assays are useful to monitor the emergence and persistence of drug-resistant HIV-1 variants in subjects infected with HIV-1 subtypes A and D.

Persistence of nevirapine in breast milk and plasma of mothers and their children after single-dose administration.

OBJECTIVES: Nevirapine is widely used in the developing world for the prevention of mother-to-child transmission (PMTCT) of HIV. A single mutation in the HIV genome is sufficient to lead to significant nevirapine resistance. Persistence of low-level drug concentrations in body compartments can foster resistance formation. In this study, concentration-time courses of nevirapine after single-dose administration were analysed over an extended post-partum period. PATIENTS AND METHODS: Breast milk and plasma samples of 62 HIV-positive Ugandan mother-child pairs who had received single-dose nevirapine were collected at delivery and 1, 2 and 6 weeks post-partum. Nevirapine concentrations were quantified by LC/tandem-mass-spectrometry using a quantification limit of 15 ng/mL, and a population pharmacokinetic (PK) analysis was performed. RESULTS: Concentration-time profiles in breast milk, maternal plasma and child plasma showed similar shapes. At week 1, median nevirapine concentrations were 164 ng/mL in maternal plasma, 114 ng/mL in breast milk and 183 ng/mL in child plasma. The population PK model predicted nevirapine concentrations>10 ng/mL (IC50 for nevirapine) for 13 days in breast milk, 14 days in maternal plasma and 18 days in child plasma in 80% of the samples. CONCLUSIONS: Nevirapine concentrations were present for 2-3 weeks in the three compartments. The concentrations are probably sufficiently high to protect most breastfed children from HIV transmission during the first 2 weeks. The long presence of slowly decreasing levels of nevirapine is likely to induce resistance formation. Post-natal addition of antiretrovirals for 1 week only, as recommended in the current PMTCT guidelines, will not suffice to avoid nevirapine resistance formation.

Male involvement in PMTCT services in Mbeya Region, Tanzania.

Throughout all stages of programmes for the prevention of mother-to-child-transmission of HIV (PMTCT), high dropout rates are common. Increased male involvement and couples' joint HIV counselling/testing during antenatal care (ANC) seem crucial for improving PMTCT outcomes. Our study assessed male attitudes regarding partner involvement into ANC/PMTCT services in Mbeya Region, Tanzania, conducting 124 individual interviews and six focus group discussions. Almost all respondents generally supported PMTCT interventions. Mentioned barriers to ANC/PMTCT attendance included lacking information/knowledge, no time, neglected importance, the services representing a female responsibility, or fear of HIV-test results. Only few perceived couple HIV counselling/testing as disadvantageous. Among fathers who had refused previous ANC/PMTCT attendance, most had done so even though they were not perceiving a disadvantage about couple counselling/testing. The contradiction between men's beneficial attitudes towards their involvement and low participation rates suggests that external barriers play a large role in this decision-making process and that partner's needs should be more specifically addressed in ANC/PMTCT services.