Sónia Centeno-Lima

A fatal case of human babesiosis in Portugal: molecular and phylogenetic analysis

We report the first case of human babesiosis in Portugal. A 66‐year‐old splenectomized man was admitted to a Lisbon hospital after 1 week of fever, abdominal pain, anorexia and nausea. A high parasitaemia (30%) of Babesia parasites was found in Giemsa‐stained blood smears and, despite treatment, the patient died several weeks later of renal failure. Ethylenediaminetetraacetic acid blood samples were processed for polymerase chain reaction (PCR) and reverse line blot hybridization to confirm and characterize the Babesia infection. The amplified PCR product was cloned and subsequently sequenced. Molecular analysis showed that the infection was caused by Babesia divergens and that other blood parasites were not involved. Phylogenetic analysis showed that the 18 S ribosomal RNA gene sequence was similar to three other European isolates of B. divergens. In view of the high risk for splenectomized individuals, strict measures should be taken to avoid tick bites.

Intestinal parasites in dogs and cats from the district of Évora, Portugal.

Intestinal parasites, both helminths and protozoa, are commonly found in domestic animals, and the possible transmission of enteric parasites from dogs and cats to humans may constitute a global potential health risk worldwide. In the present study, we analysed 148 stool samples from dogs (n=126) and cats (n=22) collected from animal shelters and veterinary clinics, in the district of Évora, Portugal. Microscopic examination confirmed that Giardia was the most frequent parasite in the studied population (34/148; 23%). Other parasites such as Ancylostoma sp., Isospora spp., Toxocara, Trichuris spp., Toxascaris and Toxoplasma were also found. Furthermore, molecular characterization of Giardia duodenalis analysis targeting the small subunit ribosomal RNA (ssu-rRNA) was performed revealing the presence of host-specific (C and D) and zoonotic assemblages (A and B). This work points out to the importance of protozoan parasites in companion animals, and reanalyses the need for parasite prophylaxis.

Evidence for a discrete evolutionary lineage within Equatorial Guinea suggests that the tsetse fly Glossina palpalis palpalis exists as a species complex

Tsetse flies of the palpalis group are major vectors of Human African Trypanosomiasis in Africa. Accurate knowledge of species identity is essential for vector control. Here, we combine ribosomal internal transcribed spacer 1 (ITS1), mitochondrial Cytochrome Oxidase 1 (COI) and microsatellites to determine the population structure and phylogenetic relations of Glossina p. palpalis in Equatorial Guinea. CO1 sequence data suggest that G. p. palpalis in Equatorial Guinea is a distinct subspecies from previously described G. p. palpalis in West Africa and Democratic Republic of Congo. Glossina p. palpalis in Equatorial Guinea and DRC share a common ancestor which diverged from West African G. p. palpalis around 1.9 Ma. Previous ITS1 length polymorphism data suggested the possible presence of hybrids in Equatorial Guinea. However, ITS1 showed incomplete lineage sorting compared with clearly defined COI groups, and data from 12 unlinked microsatellites provided no evidence of hybridization. Microsatellite data indicated moderate but significant differentiation between the populations analysed (Rio Campo, Mbini and Kogo). Moreover, unlike previous studies of G. p. palpalis, there was no evidence for heterozygote deficiency, presence of migrants or cryptic population structure. Variance effective population size at Rio Campo was estimated at 501–731 assuming eight generations per year. This study of the population genetics of G. p. palpalis in central Africa provides the first estimate of genetic differentiation between geographically separated G. p. palpalis populations.

Infection by Intestinal Parasites, Stunting and Anemia in School-Aged Children from Southern Angola

Introduction Intestinal parasites are responsible for morbidity in children worldwide, especially in low income countries. In the present study we determine the prevalence of intestinal parasites and explore its association with anemia and stunting in school-aged children. Methods A cross-sectional study was conducted from September to October 2010 enrolling 328 children attending the primary school in Lubango, the second largest city after the capital Luanda. Stool samples were collected for parasite detection through microscopy and molecular identification of Entamoeba histolytica and Entamoeba dispar. Stunting was assessed using the z-scores of height for age and hemoglobin concentration was determined using a portable hemoglobin analyzing system. Results The global prevalence of pathogenic intestinal parasites was 44.2%, the most common being Ascaris lumbricoides (22.0%), Giardia lamblia (20.1%) and Hymenolepis nana (8.8%). Molecular detection revealed that 13.1% of the children carried E. dispar and 0.3% were infected with E. histolytica. The prevalence of stunting (mild to severe) was 41.5%. Stunting was more frequent in older children (p = 0.006, OR = 1.886), while anemia was more frequent in younger children (p = 0.005, OR = 2.210). The prevalence of anemia was 21.6%, and we found a significant association with infection by H. nana (p = 0.031, OR = 2.449). Conclusions This is one of the few published studies reporting intestinal parasites infection, nutritional status and anemia in children from Angola. Furthermore, the present work highlights the importance of regular intestinal parasites screening in children.

Etiology of diarrhea in children younger than 5 years attending the Bengo General Hospital in Angola

Background: Diarrheal disease is among the leading causes of death in children younger than 5 years, especially in developing countries. The aim of this study was to investigate the most frequent etiological agents of diarrhea and its associated factors in children younger than 5 years attending the Bengo General Hospital in Angola. Methods: From September 2012 through December 2013, stool samples were collected from 344 children presenting with diarrhea to investigate the presence of viral, bacterial and parasitic agents. Relevant sociodemographic and clinical data were obtained from parents and caregivers. Results: An enteric pathogen was detected in 66.6% of stool samples: Cryptosporidium spp. (30.0%), rotavirus (25.1%), Giardia lamblia (21.6%), diarrheagenic Escherichia coli (6.3%), Ascaris lumbricoides (4.1%), adenovirus (3.8%), Strongyloides stercoralis (3.5%), astrovirus (2.6%), Hymenolepis nana (1.7%), Entamoeba histolytica/dispar (0.9%), Taenia spp. (0.6%), Trichuris trichiura (0.3%) and Entamoeba histolytica (0.3%). Children younger than 12 months were more frequently infected with Cryptosporidium spp. compared with older children (age: 12–59 months), independently of sex, season, lethargy and wasting [odds ratio (OR): 3.5, 95% confidence interval (95% CI): 2.0–6.2]. Age (OR: 5.0, 95% CI: 2.6–9.3), vomiting (OR: 2.7, 95% CI: 1.5–4.8) and type of admission (inpatients, OR: 0.5, 95% CI: 0.3–0.9) were significantly associated with rotavirus infection. Conclusions: This study demonstrates high rates of infection with an enteric pathogen, particularly in children younger than 12 months, emphasizing the need to address diarrheal disease in this age group.

Giardia duodenalis and soil-transmitted helminths infections in children in São Tomé and Príncipe: do we think Giardia when addressing parasite control?

Giardia duodenalis prevalence is commonly as high as soil-transmitted helminths (STH), nevertheless is not considered for large-scale chemotherapy through mass drug administration (MDA) due to its short incubation period and frequent reinfections, its control being associated to improving access to water and sanitation. A study enrolling 444 children attending preschools was conducted in May 2011 during a deworming campaign. Faecal samples were obtained and analysed through microscopy of wet mounting and after Kato-Katz and formol-ether concentration techniques. The majority of children were infected with at least one pathogenic parasite (86.7%, 385 of 444). Ascaris lumbricoides and Trichuris trichiura (56.3%, 250 of 444 and 52.5%, 233 of 444, respectively) were the most frequent parasites followed by G. duodenalis infecting 41.7% (185 of 444) of the children. The present work aimed at obtaining updated information concerning intestinal parasite infections in children attending preschools in São Tomé and Príncipe and to contribute for the adequate management of the enteric infections.

Studies in a co-infection murine model of Plasmodium chabaudi chabaudi and Leishmania infantum: interferon-gamma and interleukin-4 mRNA expression.

This work aimed to study the T helper type 1/2 (Th1/Th2) cytokine profile in a co-infection murine model of Plasmodium chabaudi chabaudi and Leishmania infantum. Expression of interferon-gamma (IFN-gamma) and interleukin-4 (IL-4) was analyzed, in spleen and liver of C57BL/6 mice, by reverse transcriptase-polymerase chain reaction. High levels of IFN-gamma expression did not prevent the progression of Leishmania in co-infected mice and Leishmania infection did not interfere with the Th1/Th2 switch necessary for Plasmodium control. The presence of IL-4 at day 28 in co-infected mice, essential for Plasmodium elimination, was probably a key factor on the exacerbation of the Leishmania infection.

Angola's 2013 dengue outbreak: clinical, laboratory and molecular analyses of cases from four Portuguese institutions.

INTRODUCTION Dengue virus (DENV) is the arbovirus with the widest impact on human health. In Africa in general, and in Angola in particular, the epidemiology and public health impact of DENV is far from clear. However, rapid population growth, unplanned urbanization, increased international travel, and the presence of virus major vector (Aedes aegypti) in the country suggest that DENV transmission may occur. METHODOLOGY In parallel to the occurrence of a dengue outbreak affecting the capital of Angola, between March and July 2013 four Portuguese institutions diagnosed dengue infection in 146 individuals returning to Portugal. Clinical presentation, laboratory findings, and molecular analyses of partial viral genomic segments were performed. RESULTS The mean age of the individuals included in this study was 42 years old, the majority being men of Portuguese nationality, reporting various lengths of stay in Angola. Fever was the most reported clinical sign, being frequently associated (61.0%) with myalgia and headache. Hematological values, including hematocrit, white-blood cell and platelets counts, correlated with the absence of severe or complicated cases, or coagulation disorders. No deaths were observed. Viral NS1 was detected in 56.2% of the samples, and all NS1 negative cases had anti-dengue IgM antibodies. RT-PCR indicated the presence of DENV1, which was confirmed by phylogenetic analysis of 25 partial NS5 viral sequences. CONCLUSION The DENV cases analyzed conformed to classical and uncomplicated dengue, caused by the suggested exclusive circulation of a genetically homogeneous DENV1 of genotype III, apparently with a single origin.

Molecular and Clinical Characterization of Giardia duodenalis Infection in Preschool Children from Lisbon, Portugal

Giardia duodenalis is the most prevalent intestinal protozoan infection especially in children. In Portugal scarce data are available relative to this infection in preschoolers. The present study was conducted from April to July 2009 in public preschools in Lisbon enrolling 316 children. Stool examination was performed through microscopy. Molecular analysis was conducted in all positive samples for G. duodenalis in order to determine the assemblage and subassemblage of this parasite. Eight of the preschoolers studied children (2.5%, 8/316) were infected with G. duodenalis. Additionally the brother of one of the infected children was also infected. Genotyping analysis targeting ssu-rRNA and β-giardin loci revealed six infections with assemblage A and 3 with assemblage B. Sub-assemblage determination was possible in four of the samples, with three A2 and one A3. The limited number of cases precluded an association of a determined symptom with an assemblage. The data presented here show the relevance of considering G. duodenalis analysis in children with intestinal complaints even in developed countries.