Complete Genome Sequences of 17 Clinical Campylobacter jejuni Strains from Chile.

Campylobacter jejuni is the leading cause of bacterial foodborne disease worldwide. Here, we report the complete annotated genomes and plasmid sequences of 17 Campylobacter jejuni strains isolated from patients with gastroenteritis in Santiago, Chile.

Species composition, geographical distribution and seasonal abundance of the Anopheles maculipennis complex along the Upper Rhine, Germany.

Between May and September 2016, mosquitoes were collected on a biweekly basis at 55 locations with CO2-baited encephalitis vector surveillance traps along the Upper Rhine, Germany, to evaluate the species composition, geographical distribution and abundance of the Anopheles maculipennis complex, some members of this complex being considered vectors of historical malaria in Germany. A total of 2115 Anopheles maculipennis complex specimens were collected during the season, of which a sample of 1252 individuals was determined to species level by amplification of species-specific internal transcribed spacer 2 (ITS2) sequences. A total of 856 individuals of Anopheles daciae (68.37%), 394 Anopheles messeae (31.47%) and 2 Anopheles maculipennis s.s. (0.16%) were recorded. The number and proportion of A. daciae was remarkably higher in the northern meandering zone of the Upper Rhine (843 specimens, 79.90%), than in the more canalised southern furcation zone where A. messeae with 183 collected specimens represented 92.89% of 197 classified individuals. The average number of collected A. maculipennis s.l. individuals per trapping site was 38.45, equalling 0.64% of the total mosquito collection. Despite an increase in imported malaria cases, this comparatively low abundance of A. maculipennis s.l. may indicate a low risk of endemic malaria transmission by members of the A. maculipennis complex today. The proportionally dominance of A. daciae suggests that this species could be suspected the main historical vector of malaria in the Upper Rhine region. Sequence analyses of the ITS2 fragment revealed intraindividual polymorphisms within 3 of 5 diagnostic nucleotides in all specimens of A. daciae, raising the question if additional loci should be considered, to gain further insight into the taxonomical relation to A. messeae.

Laboratory exposure to Coccidioides: lessons learnt in a non-endemic country.

Coccidioides is a primary pathogenic fungus, which infects humans through highly infectious arthroconidia, causing substantial morbidity including life-threatening disseminated infections. Due to the low infectious dose, laboratory personnel might become infected during diagnostic procedures. Accordingly, coccidioidomycosis is reported as the most frequent laboratory-acquired systemic mycosis worldwide. This risk is aggravated in non-endemic countries, where the diagnosis may not be suspected. We report on an inadvertent exposure of 44 persons to Coccidioides posadasii in a clinical microbiology laboratory in Chile, the measures of containment after rapid diagnosis with matrix-assisted laser desorption/ionization time-of-flight mass spectrometry, and the lessons learnt in a non-endemic setting.

Enteric multiplex PCR panels: A new diagnostic tool for amoebic liver abscess?

Invasive amoebiasis is a life-threatening infection requiring immediate detection and treatment. However, diagnosis is challenging because conventional methods such as light microscopy and serology are unreliable. Molecular techniques are therefore considered the new diagnostic reference standard, but most of the developed assays are research tools and not widely available. Recently commercial multiplex PCR panels have been introduced which permit the simultaneous detection of multiple enteric pathogens including Entamoeba histolytica in stool samples. Our report demonstrates for the first time that these new assays might also serve as a rapid tool to diagnose amoebic liver abscess in patients with cystic focal liver lesions.

One Health in Practice: A Pilot Project for Integrated Care of Zoonotic Infections in Immunocompromised Children and Their Pets in Chile.

Although pets provide physiological and psychological benefits to their owners, they are a potential source of zoonotic infections, especially for vulnerable individuals such as immunocompromised patients. During 1 year, we therefore performed a pilot project, which included 32 immunocompromised Chilean children and their family pets (35 dogs and 9 cats) with the aim of detecting, treating and preventing zoonotic infections. Children were examined by Infectious Diseases paediatricians and demographical and clinical information related to zoonotic infections were recorded. Pets were examined and sampled by veterinarians, who also administered missing routine vaccines and anti-parasitics. During family visits, all members were informed and educated about zoonoses and a satisfaction survey was performed. Visits also included vector control and indoor residual spraying with pyrethroids. Children were re-examined and re-tested according to the findings of their pets, and all detected zoonotic infections were treated both in children and pets. Physical examination revealed abnormalities in 18 dogs (51.4%) and three cats (33.3%). Twenty-eight (63.6%) of the pets were diagnosed with a zoonotic pathogen, and seven (15.9%) with a facultative pathogen. Most zoonotic agents were isolated from the pet's external ear and intestine. Bacteria with the highest pathogenic potential were Campylobacter jejuni and Brucella canis. In two children and their respective pets, the same zoonotic diseases were diagnosed (toxocariasis and giardiasis). Arthropods serving as potential vectors of zoonotic infections were found in 49% of dogs and 44% of cats. The pilot project was positively evaluated by the participating families. Our pilot project confirmed that pets are reservoir for various zoonotic agents in Chile and that the implementation of an integrated multidisciplinary programme was a valuable tool to prevent, diagnose and treat such zoonotic infections in vulnerable patients such as immunocompromised children.

Molecular typing of Giardia duodenalis isolates from German travellers.

Giardia duodenalis isolates from German travellers returning from tropical areas were characterised by PCR amplification and sequencing of fragments of the beta-giardin (bg), glutamate dehydrogenase (gdh) and triose phosphate isomerase (tpi) genes. Assignment of isolates to specific G. duodenalis assemblages was found to differ according to the marker used. Indeed, at the bg locus, assemblages A and B were identified, with a higher prevalence of the former over the latter, whereas at the tpi and gdh loci, most samples were classified as assemblage B. In agreement with previous studies, sequence analysis showed that assemblage B isolates have a higher genetic polymorphism than assemblage A isolates, and novel variants were described. The degree of polymorphism was shown in a graphical representation of the polymorphic sites generating a novel sequence, the heterogeneous positions common to assemblages A and B (double peaks), that may represent mixed assemblage infection and the heterogeneous positions detected at random sites. Notably, assemblage D, which is considered to be adapted to dogs, was found at the gdh locus in two samples originating from southern Asia, as novel genotypes. By comparing the geographical origin of the infected cases and the number of German travellers visiting the areas considered, India and west Africa appeared to be the areas associated to the highest risk of acquiring Giardia infection. The analysis of the geographical distribution of the genotypes did not suggest any particular geographical clustering pattern, but it may be useful to evaluate these results with a higher number of isolates. Most of the samples typed at the three markers could not be assigned unequivocally to either assemblage A or B, and this was confirmed also by a real-time PCR assay, using a set of assemblage-specific primers. The results of this study reinforce the notion that genetic exchanges and allelic sequence heterogeneity represent major obstacles towards understanding the epidemiology of giardiasis and that exposure to Giardia parasites in endemic areas often results in mixed infections in returning travellers.

Acute fascioliasis--clinical and epidemiological features of four patients in Chile.

Because of its infrequent and protean presentation and the lack of clinical data, the management of acute infections with the foodborne trematode Fasciola hepatica is challenging. We report four serologically confirmed cases that illustrate our experience with this parasitic infection in Chile. All patients were adults presenting with upper abdominal pain. Other symptoms included fever, nausea/vomiting, and cutaneous manifestations. In all cases, marked eosinophilia was present. All patients lived in an urban environment, and three reported the consumption of raw watercress. Computed tomography (CT) scans showed hypodense hepatic lesions, whereas ultrasonography findings were unremarkable. One patient suffered portal vein thrombosis, which might be a rare complication of acute fascioliasis. All patients were successfully treated with triclabendazole. Our case series demonstrates that patients with acute fascioliasis typically present with a combination of upper abdominal pain, marked eosinophilia, and hypodense hepatic lesions on CT imaging. Diagnosis should be confirmed by serological investigation. A history of recent consumption of raw watercress is an important finding, but in some patients the source of infection remains obscure.

Giardiasis in kindergartens: prevalence study in Berlin, Germany, 2006.

In Germany, an increase of notified giardiasis was observed between 2002 (n = 3,101) and 2007 (n = 3,651) with 62% of cases acquired in Germany. The highest incidence was reported in 1- to 5-year-olds (2001-2007, 11.5/100,000 on average) and Berlin is one of the most affected states (17.5/100,000). We performed a cross-sectional study in five Berlin kindergartens differing in socioeconomic status and migrant proportion in order to estimate the prevalence and investigate routes of transmission among children under 6 years of age. Stool samples were screened for Giardia lamblia and Cryptosporidium parvum using microscopical and antigen detection. Giardia-positive samples underwent PCR and subtyping. Two hundred two children participated (mean age 3.4 years). We found three girls (1.5%) who tested positive for G. lamblia genotype-A3, all clustering in one kindergarten (prevalence 5.5%). No common source was identified. Two children were symptomatic. Possible dog-to-child transmission was established for one of the symptomatic cases. All contact-tracing results were negative. Other microscopically detected parasites (Blastocystis hominis, Endolimax nana cysts, Entamoeba coli cysts and Iodamoeba bütschlii cysts) were found in 4% of children. In summary, a substantial level of Giardiasis may be prevalent in some of Berlin's kindergartens, despite standard hygienic measures being followed. The relatively high prevalence in one kindergarten indicates the need for further studies to identify risk factors for children, which may help to guide possible interventions and strategies. Giardiasis should be considered as a differential diagnosis in children with unclear gastrointestinal symptoms. Additional education and training on proper toilet and food hygiene may further reduce the possibility of child-to-child transmission.

Evaluation of a new point-of-care test for influenza A and B virus in travellers with influenza-like symptoms.

Point-of-care (POC) tests for influenza facilitate clinical case management, and might also be helpful in the care of travellers who are at special risk for influenza infection. To evaluate influenza POC testing in travellers, a new assay, the ImmunoCard STAT! Flu A and B, was used to investigate travellers presenting with influenza-like symptoms. Influenza virus infection was diagnosed in 27 (13%) of 203 patients by influenza virus-specific PCR and viral culture. The POC test had sensitivity and specificity values of 64% and 99% for influenza A, and 67% and 100% for influenza B, respectively. Combined sensitivity and specificity were 67% and 99%, respectively, yielding positive and negative predictive values of 95%, and positive and negative likelihood ratios of 117 and 0.34, respectively. The convenient application, excellent specificity and high positive likelihood ratio of the POC test allowed rapid identification of influenza cases. However, negative test results might require confirmation by other methods because of limitations in sensitivity. Overall, influenza POC testing appeared to be a useful tool for the management of travellers with influenza-like symptoms.

Epidemiological and clinical features of travel-associated cryptosporidiosis.

Data concerning the clinical and epidemiological features of travel-associated cryptosporidiosis are lacking. In order to investigate the impact of this disease on travellers' health, a retrospective study was conducted at the Institute of Tropical Medicine, Berlin. In total, 57 cryptosporidial infections were identified between 2000 and 2004, resulting in a prevalence of 2.9% in patients with travel-associated diarrhoea. Travel to south-central Asia, especially India, was associated with a higher prevalence of infection than was travel to other destinations. Clinically, the disease resembled giardiasis, but fever and arthralgias seemed to occur more frequently.

Evaluation of seven commercial antigen detection tests for Giardia and Cryptosporidium in stool samples.

Stool samples from patients with abdominal symptoms were used to evaluate different copro-diagnostic assays for the detection of Giardia and Cryptosporidium. Results from microscopical examination following conventional stool concentration and direct fluorescent-antibody methods were compared with various commercially available immunochromatographic and enzyme immunoassays. Of 220 samples, 45 were positive for Giardia and 17 for Cryptosporidium. For Giardia, the sensitivities obtained by Ridascreen Giardia, Rida Quick Giardia, Rida Quick Combi and Giardia-Strip were 82%, 80%, 80% and 44%, respectively. For Cryptosporidium, the sensitivities obtained by Rida Quick Cryptosporidium, Ridascreen Cryptosporidium, Rida Quick Combi and Cryptosporidium-Strip were 88%, 82%, 82% and 75%, respectively. The specificity of all tests was > or = 98%. Other intestinal parasites were present in 68 samples, but cross-reactions with other protozoan or helminthic parasites were not observed. Overall, the copro-antigen assays were less time-consuming and easier to perform, but were less sensitive than conventional microscopical methods. Thus, these tests might be a useful addition to, but not a substitute for microscopical methods in the diagnosis of travel-associated giardiasis and cryptosporidiosis.

Imported leishmaniasis in Germany 2001-2004: data of the SIMPID surveillance network.

Leishmaniasis is a rare, non-notifiable disease in Germany. Epidemiological and clinical data, therefore, are scarce. Most infections seen in Germany are contracted outside the country. The German surveillance network for imported infectious diseases (Surveillance Importierter Infektionen in Deutschland, or SIPMID) recorded 42 cases of imported leishmaniasis (16 visceral, 23 cutaneous, and 3 mucocutaneous) from January 2001 to June 2004. Although most infections were acquired in European Mediterranean countries, the risk of infection was highest for travelers to Latin America. HIV coinfection was observed significantly more often in patients with visceral leishmaniasis than in patients with cutaneous/mucocutaneous leishmaniasis (31 vs. 4%, p=0.02). The median time to a definitive diagnosis was 85 days in cases of visceral leishmaniasis and 61 days in cases of cutaneous/mucocutaneous leishmaniasis, reflecting the unfamiliarity of German physicians with leishmanial infections. Visceral leishmaniasis was treated most frequently with amphotericin B, whereas cutaneous/mucocutaneous leishmaniasis was treated with a variety of local and systemic therapies. The findings presented here should serve to increase awareness as well as improve clinical management of leishmaniasis in Germany.

Age as a risk factor for severe manifestations and fatal outcome of falciparum malaria in European patients: observations from TropNetEurop and SIMPID Surveillance Data.

Previous studies have indicated that age is a risk factor for severe falciparum malaria in nonimmune patients. The objectives of this study were to reevaluate previous findings with a larger sample and to find out how strongly clinical outcomes for elderly patients differ from those for younger patients. Results of adjusted analyses indicated that the risks of death due to falciparum malaria, of experiencing cerebral or severe disease in general, and of hospitalization increased significantly with each decade of life. The case-fatality rate was almost 6 times greater among elderly patients than among younger patients, and cerebral complications occurred 3 times more often among elderly patients. Antimalarial chemoprophylaxis was significantly associated with a lower case-fatality rate and a lower frequency of cerebral complications. Women were more susceptible to cerebral complications than were men. Our study provides evidence that falciparum malaria is more serious in older patients and demonstrates that clinical surveillance networks are capable of providing quality data for investigation of rare events or diseases.

Resistance analyses in HIV infected patients with a history of multiple antiretroviral treatment regimens.

OBJECTIVE: To assess HIV-1 isolate based resistance profiles from extensively pretreated patients and effects of a resistance guided switch of antiretroviral therapy. METHODS: In a prospective study phenotypic and genotypic resistance analyses were performed on HIV infected individuals with failure of the current therapy and history of at least three antiretroviral regimens. Antiretroviral therapy was changed according to the results. Viral load and CD4 lymphocyte counts were measured at baseline, after 10 (SD 2), and 24 (2) weeks. RESULTS: All patients (n=52) failed their actual regimen. Currently versus ever previously taking the specific drug, resistance associated mutations and phenotypic resistance to AZT and 3TC were found in over 80% of individuals; resistance to DDI and D4T was detected in less than 10% of cases. A resistance guided switch of therapy was followed by a median decrease of viral load of 0.5 log10 units after 24 weeks. Individuals resistant to two or more drugs compared with patients with resistance to less than two drugs of ongoing treatment, were switched to a regimen containing DDI, D4T, and a PI or NNRTI. After 10 (SD 2) weeks viral load decrease was pronounced in patients with resistance to at least two drugs in the previous regimen. CONCLUSIONS: Among different RTI, the profile of clinically relevant resistance indicates pronounced differences when looking at separate drugs. Regarding virological response, in the context of available drugs, resistance tested with currently used methods is of limited value in extensively pretreated patients and seems to have its value primarily in first or second switch of therapy.

Dual microsporidial infection with Encephalitozoon cuniculi and Enterocytozoon bieneusi in an HIV-positive patient.

This report describes the first dual microsporidial infection with Encephalitozoon cuniculi and Enterocytozoon bieneusi in an HIV-positive patient. In view of clinical and epidemiological findings, our E. cuniculi isolate was deduced to be of the dog strain. The patient's occupational involvement with dogs indicates that canines should be considered as a reservoir of human infections for both microsporidial species. Furthermore, our report provides detailed clinical and radiological information on a rare case of a symptomatic pulmonary infection by E. cuniculi and its improvement after treatment with albendazole.

Differences between American and German medical students and physicians in knowledge of infectious diseases.

Unlike in the USA and many other countries, the discipline of infectious diseases is not established as a specialty or a subspecialty in Germany. In order to assess the impact of this structural difference, the knowledge of clinical infectious diseases was compared among 64 German and 82 American physicians and medical students using a 33-question, multiple-choice questionnaire. Participants in the USA had significantly better scores, and, among them, the number of correct answers increased steadily along with clinical experience. In contrast, among the German participants only a minor increase in knowledge was observed after the first 2 years of postgraduate training. The superior knowledge of infectious diseases demonstrated by physicians and medical students in the USA is presumably due to better training in this field during medical school and residency. Additionally, repeated exposure to infectious disease consultation in daily practice is likely to provide continuous medical education. The results of this study may provide a rationale for establishing infectious diseases as a recognized medical subspecialty in Germany.