Malaria screening: what is the contribution of molecular biology?

INTRODUCTION: According to the World Health Organization, Microscopy is the gold standard for diagnosing malaria. However, the performance of this examination depends on the experience of the microscopist and the level of parasitemia. Thus, molecular biology detection of malaria could be an alternative technique. AIM: evaluate the contribution of molecular biology in detecting imported malaria. METHODS: This was a descriptive, prospective study, including all students, from the Monastir region, and foreigners, from countries endemic to malaria. The study period was from September 2020 to April 2021. Each subject was screened for malaria by three methods: direct microscopic detection of Plasmodium, detection of plasmodial antigens, and detection of plasmodial DNA by nested PCR. RESULTS: Among the 127 subjects screened, only one had a positive microscopic examination for Plasmodium falciparum. Among the 126 subjects with a negative microscopic examination, twelve students had a positive nested PCR result, i.e. 9.5%. Molecular sequencing allowed the identification of ten isolates of Plasmodium falciparum, one Plasmodium malariae and one Plasmodium ovale. Our study showed that the results of nested PCR agreed with those of microscopy in 90.6% of cases. CONCLUSION: Nested PCR seems more sensitive for the detection of low parasitemias. Hence the importance of including molecular biology as a malaria screening tool to ensure better detection of imported cases.

Imported schistosomiasis in travelers: Experience from a referral tropical medicine unit in Barcelona, Spain.

BACKGROUND: Acute schistosomiasis occurs most often in travelers to endemic regions. The aim of the study is to describe the epidemiological, clinical and parasitological characteristics of patients with schistosomiasis acquired during an international travel. METHODS: Observational retrospective study including all travel-related schistosomiasis cases seen at the International Health Unit Vall d'Hebron-Drassanes (Barcelona, Spain) from 2009 to 2022. Diagnosis of schistosomiasis was defined by the presence of Schistosoma eggs in stools or urine or the positivity of a serological test. We collected demographic, epidemiological, clinical, parasitological, and therapeutic information. RESULTS: 917 cases of schistosomiasis were diagnosed, from whom 96 (10.5 %) were travel-related. Mean age of the patients was 34.9 years, and 53.1 % were women. Median duration of the travel was 72 days, and geographical areas where travelers had contact with fresh water were Africa (82.3 %), Asia (12.5 %), and South America (5.2 %). Twenty (20.8 %) patients reported having had some clinical symptom, being gastrointestinal symptoms the most frequent. Two patients developed the classical Katayama syndrome. In eleven (11.5 %) cases eggs were observed in urine or feces samples, and 85 (88.5 %) cases were diagnosed by a positive serology. Ninety-one (94.8 %) patients received treatment with praziquantel with different therapeutic schemes. The two patients with Katayama syndrome received concomitant treatment with corticosteroids. CONCLUSIONS: Schistosomiasis in travelers represented 10 % of the overall schistosomiasis cases in our center. Increasing the awareness in the pre-travel advice and implementing specific screening in those travelers at risk (long travelers, contact with fresh water) could reduce the incidence and associated morbidity in this group.

Clinical and epidemiological features of imported loiasis in Beijing: a report from patients returned from Africa.

BACKGROUND: Loiasis is one of the significant filarial diseases for people living in West and Central Africa with wide endemic area but is not seen in China. As economy booms and international traveling increase, China faces more and more imported parasitic diseases that are not endemic locally. Loiasis is one of the parasitic diseases that enter China by travelers infected in Africa. The better understanding of the clinical and laboratory features of loa loa infection will facilitate the diagnosis and treatment of loiasis in China. METHODS: The study targeted travelers who were infected with L. loa in endemic Africa regions and returned to Beijing between 2014 and 2023. Epidemiological, clinical, and biological data as well as treatment of these patients were collected. RESULTS: Total 21 cases were identified as L. loa infection based on their typical clinical manifestations and parasite finding. All cases had a history of travel to Africa for more than 6 months, most of them are the construction workers dispatched to West Africa with outdoor activities. Calabar swelling (n = 19; 90.5%) and pruritus (n = 11; 52.4%) were among the most common clinical symptoms followed by muscle pain (n = 7; 33.3%) and skin rash (n = 2; 9.5%). The adult worms were observed in the eyelid or subconjunctiva (n = 2; 9.5%) and subcutaneous tissues (n = 2; 9.5%). Although all patients presented with a high eosinophil count (> 0.52 × 109/L), only two cases displayed microfilariae in fresh venous blood and positive for filarial antigen. A cut section of adult worm was observed through biopsy on a skin nodule surrounded by lymphocytes, plasma cells and eosinophils. All subjects were positive in PCR targeting L. loa ITS-1. The constructed phylogenetic tree based on the amplified ITS-1 sequences identified their genetical relation to the L. Loa from Africa. All patients treated with albendazole and diethylcarbamazine were recovered without relapse. CONCLUSION: This study provides useful information and guideline for physicians and researchers in non-endemic countries to diagnose and treat loiasis and L. loa infections acquired from endemic regions.

Imported cutaneous leishmaniasis: molecular investigation unveils Leishmania major in Bangladesh.

BACKGROUND: The main clinical forms of leishmaniasis in Bangladesh are visceral leishmaniasis and post-kala-azar dermal leishmaniasis, which are caused by Leishmania donovani. Imported cutaneous leishmaniasis (CL) is emerging globally due mainly to increased human mobility. In recent years, several imported CL cases have also been reported in Bangladesh. Sporadic atypical cases of CL can be challenging for diagnosis and clinical management, while occurrence of infection on a frequent basis can be alarming. We report of a case of a Bangladeshi temporary-migrant worker who, upon return, presented development of skin lesions that are characteristic of CL. METHODS: A serum sample was collected and tested with an rK39 immunochromatographic test. Nucleic acid from skin biopsy derived culture sample was extracted and screened with a real-time PCR assay which targets the conserved REPL repeat region of L. donovani complex. The internal transcribed spacer 2 region of the ribosomal RNA gene cluster was amplified and sequenced. RESULTS: The suspect had a history of travel in both CL and VL endemic areas and had a positive rK39 test result. Based on clinical presentation, travel history and demonstration of the parasite in the skin biopsy, CL was diagnosed and the patient underwent a combination therapy with Miltefosine and liposomal amphotericin B. While typical endemic species were not detected, we identified Leishmania major, a species that, to our knowledge, has never been reported in Bangladesh. CONCLUSIONS: Proper monitoring and reporting of imported cases should be given careful consideration for both clinical and epidemiological reasons. Molecular tests should be performed in diagnosis to avoid dilemma, and identification of causative species should be prioritized.

[Laboratory diagnosis of an imported case of visceral leishmaniasis in Henan Province].

OBJECTIVE: To analyze the epidemiological and clinical data of an imported case of visceral leishmaniasis in Henan Province, and explore the method of laboratory diagnosis of kala-azar. METHODS: The epidemiological and clinical data of an imported visceral leishmaniasis patient were analyzed. Leishmania donovani bodies in bone marrow smears were observed microscopically. The antibody was detected by using rK39 dipstick test strips. Two pairs of specific primers, K13A-K13B and LITSR-L5.8S, were used to amplify kinetoplast DNA and internal transcribed spacer of rDNA of the parasite, respectively. RESULTS: The patient had been in the epidemic area of visceral leishmaniasis, and had symptoms such as irregular fever, splenomegaly, pancytopenia, and inversed ratio of albumin and globulin. The amastigotes of L. donovani were found in the bone marrow smears, and rK39 test strip was positive, and the PCR products of K13A-K13B and LITSR-L5.8S were 87 bp and 285 bp respectively. The similarities of the two fragment sequences to the corresponding sequences of L. donovani were 94% and 100%, respectively. CONCLUSIONS: The case is diagnosed as visceral leishmaniasis according to the epidemiological data, clinical manifestations and laboratory test results of the patient, and the pathogen is L. donovani.

Imported cases of malaria in Spain: observational study using nationally reported statistics and surveillance data, 2002-2015.

BACKGROUND: Malaria was eliminated in Spain in 1964. Since then, more than 10,000 cases of malaria have been reported, mostly in travellers and migrants, making it the most frequently imported disease into this country. In order to improve knowledge on imported malaria cases characteristics, the two main malaria data sources were assessed: the national surveillance system and the hospital discharge database (CMBD). METHODS: Observational study using prospectively gathered surveillance data and CMBD records between 2002 and 2015. The average number of hospitalizations per year was calculated to assess temporal patterns. Socio-demographic, clinical and travel background information were analysed. Bivariate and multivariable statistical methods were employed to evaluate hospitalization risk, fatal outcome, continent of infection and chemoprophylaxis failure and their association with different factors. RESULTS: A total of 9513 malaria hospital discharges and 7421 reported malaria cases were identified. The number of reported cases was below the number of hospitalizations during the whole study period, with a steady increase trend in both databases since 2008. Males aged 25-44 were the most represented in both data sources. Most frequent related co-diagnoses were anaemia (20.2%) and thrombocytopaenia (15.4%). The risks of fatal outcome increased with age and were associated with the parasite species (Plasmodium falciparum). The main place of infection was Africa (88.9%), particularly Equatorial Guinea (33.2%). Most reported cases were visiting friends and relatives (VFRs) and immigrants (70.2%). A significant increased likelihood of hospitalization was observed for children under 10 years (aOR:2.7; 95% CI 1.9-3.9), those infected by Plasmodium vivax (4.3; 95% CI 2.1-8.7) and travellers VFRs (1.4; 95% CI 1.1-1.7). Only 4% of cases reported a correct regime of chemoprophylaxis. Being male, over 15 years, VFRs, migrant and born in an endemic country were associated to increased risk of failure in preventive chemotherapy. CONCLUSIONS: The joint analysis of two data sources allowed for better characterization of imported malaria profile in Spain. Despite the availability of highly effective preventive measures, the preventable burden from malaria is high in Spain. Pre-travel advice and appropriately delivered preventive messages needs to be improved, particularly in migrants and VFRs.

A rare case of visceral leishmaniasis in an immunocompetent traveler returning to the United States from Europe.

A young, healthy traveler returning to the United States presented with fever, night sweats, splenomegaly, and pancytopenia. Bone marrow biopsy revealed leishmaniasis (Leishmania infantum), likely acquired in southern France. Although many cases of endemic visceral leishmaniasis (VL) have been reported in Europe, this is a rare case of imported VL in a healthy traveler returning from Europe to the US. Despite successful initial treatment with liposomal amphotericin B (LamB), relapse occurred. Treatments for VL in immunocompetent individuals are highly effective, but relapse can occur. There is more extensive experience in endemic areas with treating relapse that may be lacking in North America. This case alerts physicians in the US that immunocompetent adults can acquire VL during brief visits to endemic areas in Europe. It is important that travelers be counseled on preventive measures. Patients should be monitored after treatment for relapse.

Status of imported malaria on Réunion Island in 2016.

BACKGROUND: Autochthonous malaria has been eliminated from Réunion in 1979. To prevent secondary transmission and re-emergence of autochthonous malaria, permanent epidemiologic and entomological surveillance and vector control measures are conducted around imported malaria cases. Results of local malaria surveillance (clinical data and results of epidemiological and entomological investigations around cases) were collected for 2013-2016 and were analysed according to historical data and to the exchanges with malaria-affected areas (estimated by airport data). RESULTS: Form 2013 to 2016, 95 imported malaria cases have been detected in Reunion Island: 42% of cases occurred in the area of repartition of Anopheles arabiensis, but Anopheles mosquitoes were present only around seven cases including one gametocyte carrier. No autochthonous or introduced case has occurred during this period. The lack of chemoprophylaxis or poor adherence was found in the majority (96%) of malaria cases between 2013 and 2016, regardless of trip type. Affinity tourism in Madagascar and Comoros was the cause of 65% of imported malaria cases. DISCUSSION: The incidence of imported malaria and the incidence rate per 100,000 travellers has continuously decreased since 2001. Now with the drastic decrease of malaria transmission in the Comoros archipelago, most of imported malaria cases in Reunion Island have been contaminated in Madagascar. Immigrants regularly resident in Reunion Island, which travel to malaria endemic countries (mainly Madagascar) to visit their friends and relatives (VFRs) represent a high-risk group of contracting malaria. VFRs, low adherence to pre-travel recommendations, in particular, the compliance on the use of chemoprophylaxis are the main drivers of imported malaria in Reunion Island. Furthermore as previously described, some general practitioners in Reunion Island are always not sufficiently aware of the official recommendations for prescriptions of prophylactic treatments. CONCLUSION: Social mobilization targeted on the Malagasy community in Reunion Island could help to decrease the burden of imported malaria in Reunion Island. Because of the low number of gametocyte carriers and the absence of an Anopheles mosquito population when most malaria cases were imported those last 4 years, the risk of the appearance of introduced malaria cases and indigenous malaria cases appears low in Reunion Island.

Changes in malaria epidemiology in Germany, 2001-2016: a time series analysis.

BACKGROUND: German surveillance data showed a sharp rise of malaria cases in 2014 and 2015 due to the increased arrival of refugees from malaria endemic countries. A time series analysis of data from 2001 to 2016 was performed in order to describe the epidemiology of imported malaria in Germany in general and of the recent increase in particular. RESULTS: In total, 11,678 malaria cases were notified between 2001 and 2016 (range 526-1063 cases/year). Newly arriving refugees averaged 10 cases/year (1.5%) in 2001-13 and 292.5 cases/year (28.3%) in 2014-15. Plasmodium (P.) falciparum was the most frequently reported species (range 57.2-85.8%), followed by P. vivax (range during 2001-2013: 7.6-18.1%; during 2014-2015, mean 31.3%). In 2014-15, 22.3% of all P. vivax cases were refugees from Eritrea and 3.3% from other countries of the Horn of Africa; in 2015 and 2016, 19.5% were refugees from Afghanistan and Pakistan. Five P. knowlesi malaria infections were reportedly acquired in Thailand between 2012 and 2016. Total numbers of malaria notifications among native Germans and residents with migration background showed an increasing trend since 2007. Chemoprophylaxis use was reported for 24.3% (1695/6984) of cases and showed a declining trend. Native German cases took significantly more frequently chemoprophylaxis than cases with migration background (32.6% vs. 17.9%; p < 0.001). DISCUSSION/CONCLUSIONS: The steep rise in vivax malaria notifications in 2014 and 2015 was mainly due to newly arriving refugees from Eritrea but also from other countries of the Horn of Africa and South Asia. Clinicians should include malaria in their differential diagnosis in case of a febrile illness in the respective population and consider vivax malaria even if arrival to Germany dates back several months. Over the past 10 years, malaria notifications among native Germans and residents with migration background showed an increasing trend. Use of chemoprophylaxis was insufficient in both groups and deteriorating. New strategies need to be found to increase compliance to chemoprophylaxis recommendations. The surveillance provides valuable data for epidemiological assessment of imported malaria in Germany.

Uncommon and fatal case of cystoisosporiasis in a non HIV-immunosuppressed patient from a non-endemic country.

Cystoisospora belli (previously known as Isospora belli) is a tropical coccidian parasite sometimes leading to severe diarrhea in immunocompromised patients. Here we describe a fatal case of cystoisosporiasis in a non HIV-immunocompromised 71-year-old female with no recent travel history. Infection was either latent or potentially caused by the consumption of contaminated imported food from Asia. Diagnosis was made by microscopical detection of numerous C. belli oocysts in stools without specific staining. Treatment with TMP-SMZ slightly improved diarrhea within 3days, but dehydration subsequently led to acute decompensated heart failure and a fatal evolution. This report illustrates the possibility of severe cystoisosporiasis in non HIV-immunocompromised patients in a non-endemic country and highlights the risk of transmission through imported contaminated food consumption.