Dengue virus serotype 4 and chikungunya virus coinfection in a traveller returning from Luanda, Angola, January 2014.

A concurrent dengue virus serotype 4 and chikungunya virus infection was detected in a woman in her early 50s returning to Portugal from Luanda, Angola, in January 2014. The clinical, laboratory and molecular findings, involving phylogenetic analyses of partial viral genomic sequences amplified by RT-PCR, are described. Although the circulation of both dengue and chikungunya viruses in Angola has been previously reported, to our knowledge this is the first time coinfection with both viruses has been detected there.

Molecular characterization of Giardia duodenalis in children from the Cufada Lagoon Natural Park, Guinea-Bissau.

Molecular characterization of Giardia duodenalis in African countries is relatively scarce. The global understanding of Giardia epidemiology is reinforced when more data are available from highly endemic countries. In the present study, 50 fecal samples collected from children from Guinea-Bissau were screened for Giardia infection. Amplification of the Giardia ssu-rRNA fragment was achieved for 28 samples (28/50, 56.0 %) of which 23 (23/50, 46.0 %) positive samples for Giardia were detected through microscopy. Eighteen samples previous amplified for the ssu-rRNA locus were amplified for the bg gene fragment. Sequence analysis was performed in 26 and 17 samples for the ssu-rRNA and bg gene fragment, respectively. Our results revealed a predominance of assemblage B (22/26, 84.6 %), sequences with high genetic polymorphism among isolates belonging to this assemblage, making impossible the subassemblage determination. Assemblage A was identified in three isolates (3/26, 11.5 %), and our results strongly suggest that two isolates belong to subassemblage A2. This study provides information about G. duodenalis genotypes in a rural area of Guinea-Bissau and may contribute for a better understanding of giardiasis epidemiology in this country.

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.

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.

Follow-up of anti-Aspergillus IgG and IgA antibodies in bone marrow transplanted patients with invasive aspergillosis.

A total of 89 patients at risk for, or with invasive aspergillosis (IA) were recruited from bone marrow transplantation (BMT) units in two Lisbon hospitals, and followed for 2(1/2) years to monitor their immune response. Of these patients, six developed probable IA, from which five died. The presence of serum IgG or IgA antibodies against seven Aspergillus recombinant antigens was assessed in patients with IA, using an enzyme-linked immunosorbent assay (ELISA). In parallel, the serum levels of galactomannan (GM) were also monitored, using the Platelia Aspergillus kit (Sanofi Pasteur, Marnes-la-Coquette, France). Superoxide dismutase (Sod) and 94 kDa were the most immunogenic antigens for IgA, while the IgG pattern of recognition changed from patient to patient. From our results we conclude that although follow-up of antibodies against these antigens should not be used as a diagnostic method, patients with IA do produce an immune response that may influence disease outcome.