An 840-bp fragment of the 18S rRNA gene was used to identify Cryptosporidium spp. recovered from human immunodeficiency virus (HIV)-infected and -uninfected patients from Kenya, Malawi, Brazil, the United Kingdom, and Vietnam. Initial identification was by Ziehl-Neelsen acid-fast staining. Confirmation was by nested PCR, targeting the most polymorphic region of the 18S rRNA gene. Genotyping was by restriction endonuclease digestion of the PCR product followed by nucleotide sequencing. Among 63 isolates analyzed, four genotypes of Cryptosporidium were identified; 75% of the isolates were of the C. parvum human genotype, while the potentially zoonotic species were of the C. parvum bovine genotype (21.7%), the C. meleagridis genotype (1.6% [one isolate]), and the C. muris genotype (1.6% [one case]). HIV-infected individuals were more likely to have zoonotic genotypes than the HIV-uninfected individuals. Among the C. parvum group, strains clustered distinctly into either human or bovine genotypes regardless of the geographical origin, age, or HIV status of the patients. The intragenotypic variation observed in the C. parvum human genotype was extensive compared to that within the C. parvum bovine genotype group. The variation within genotypes was conserved in all geographical regions regardless of the patients' HIV status. The extensive diversity within genotypes at the 18S rRNA gene locus may limit its application to phylogenetic analyses.The use of molecular methods in the taxonomy of Cryptosporidium spp. has led to increased recognition of the diversity of the species infecting humans. The main causative agents of human cryptosporidiosis are strains of human and bovine genotypes of the species C. parvum (also called genotype 1 and genotype 2, respectively) (3). Recent reports document human infections with zoonotic species, including C. parvum dog and cat types (now renamed C. canis and C. felis, respectively) and C. meleagridis and C. muris (mainly avian and murine parasites, respectively) (4,5,11,13,24). A recently recognized Cryptosporidium cervine genotype has been identified in both immunocompromised and immunocompetent people (9). However, the prevalence and significance of the different species and genotypes in both immunocompetent and immunodeficient people are not yet clear. Experimental studies of both humans and animals suggest that diverse species and genotypes show different levels of infectivity and virulence (8). Others have pointed out that zoonotic strains of C. parvum produce more severe infections in humans than the strains found only in humans (7). Moreover, the potential reservoir hosts and transmission pathways for the novel species infecting humans are unclear (3). These issues underscore the importance of the precise identification of species recovered from humans.So far, a limited number of isolates have been typed from developing countries and especially from human immunodeficiency virus (HIV)-infected people (5,12,16,17,19,20,24), yet cryptosporidiosis is endemic in most tropical region...

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A descriptive study on rickettsiosis was conducted at the largest referral hospital in Hanoi, Vietnam, to identify epidemiological and clinical characteristics of specific rickettsiosis. Between March 2001 and February 2003, we enrolled 579 patients with acute undifferentiated fever (AUF), excluding patients with malaria, dengue fever, and typhoid fever, and serologically tested for Orientia tsutsugamushi and Rickettsia typhi. Of the patients, 237 (40.9%) and 193 (33.3%) had scrub and murine typhus, respectively, and 149 (25.7%) had neither of them (non–scrub and murine typhus [non-ST/MT]). The proportion of murine typhus was highest among patients living in Hanoi whereas that of scrub typhus was highest in national or regional border areas. The presence of an eschar, dyspnea, hypotension, and lymphadenopathy was significantly associated with a diagnosis of scrub typhus (OR = 46.56, 10.90, 9.01, and 7.92, respectively). Patients with murine typhus were less likely to have these findings but more likely to have myalgia, rash, and relative bradycardia (OR = 1.60, 1.56, and 1.45, respectively). Scrub typhus and murine typhus were shown to be common causes of AUF in northern Vietnam although the occurrence of spotted fever group rickettsiae was not determined. Clinical and epidemiological information may help local clinicians make clinical diagnosis of specific rickettsioses in a resource-limited setting.

Background: Confirmation of pulmonary tuberculosis (PTB) in young children is difficult as they seldom expectorate sputum. Aim: To compare sputa obtained by nasopharyngeal aspiration and by sputum induction for staining and culture of Mycobacterium tuberculosis. Patients and methods: Patients from Mulago Hospital, Kampala with symptoms suggestive of PTB were considered for inclusion in the study. Those with a positive tuberculin test and/or a chest radiograph compatible with tuberculosis were recruited. Infection with human immunodeficiency virus (HIV) was confirmed by duplicate enzyme-labelled immunosorbent assay or in children ,15 months by polymerase chain reaction (PCR). Direct PCR was undertaken on 82 nasopharyngeal aspirates. Results: Of 438 patients referred, 94 were recruited over a period of 5 months. Median (range) age was 48 (4-144) months. Of 63 patients tested, 69.8% were infected with HIV. Paired and uncontaminated culture results were available for 88 patients and smear results for 94 patients. Nasopharyngeal aspirates were smear-positive in 8.5% and culture-positive in 23.9%. Induced sputa were smear-positive in 9.6% and culture positive in 21.6%. Overall, 10.6% were smear-positive, 25.5% were culture-positive and 26.6% had smear and/or culture confirmed tuberculosis. Direct PCR on nasopharyngeal aspirates had a sensitivity of 62% and specificity of 98% for confirmation of culture-positive tuberculosis. Conclusions: Nasopharyngeal aspiration is a useful, safe and low-technology method for confirmation of PTB and, like sputum induction, can be undertaken in outpatient clinics.T he diagnosis of pulmonary tuberculosis (PTB) in young children is usually based on contact history, positive tuberculin test and compatible chest radiography. However, confirmation of tuberculosis by culture is important where, for example, there is difficulty in clinical diagnosis, the contact history is not known or drug sensitivities are not available. In developing countries the diagnosis of PTB is particularly difficult as the contact history is often not clear (especially where tuberculosis/human immunodeficiency virus (HIV) co-infection in the household is common) and the tuberculin test is often falsely negative. Chest radiography may be difficult to interpret, especially when there is HIV-related pulmonary disease such as lymphocytic interstitial pneumonitis (LIP). Obtaining sputum in young children who are unable to expectorate is problematic. Gastric aspiration usually requires a child to be admitted for up to 3 days and has to be undertaken in the early morning while the child is recumbent and fasted overnight.3 Alternative methods include sputum induction (SI), 4 5 laryngeal swab (LS) 6 7 and nasopharyngeal aspiration (NPA).8 9 This study aimed to compare the value of NPA with SI for confirmation of PTB. METHODSThe study was undertaken at Mulago Hospital, Kampala during the period January to June 2004. A diagnostic room was set up for SI, NPA and tuberculin testing, organised by a dedicated paediatrician, a nur...

To determine trends, mortality rates, and costs of antimicrobial resistance in invasive bacterial infections in hospitalized children, we analyzed data from Angkor Hospital for Children, Siem Reap, Cambodia, for 2007–2016. A total of 39,050 cultures yielded 1,341 target pathogens. Resistance rates were high; 82% each of Escherichia coli and Klebsiella pneumoniae isolates were multidrug resistant. Hospital-acquired isolates were more often resistant than community-acquired isolates; resistance trends over time were heterogeneous. K. pneumoniae isolates from neonates were more likely than those from nonneonates to be resistant to ampicillin–gentamicin and third-generation cephalosporins. In patients with community-acquired gram-negative bacteremia, third-generation cephalosporin resistance was associated with increased mortality rates, increased intensive care unit admissions, and 2.26-fold increased healthcare costs among survivors. High antimicrobial resistance in this setting is a threat to human life and the economy. In similar low-resource settings, our methods could be reproduced as a robust surveillance model for antimicrobial resistance.

Summaryobjectives To determine the levels of resistance to first-line tuberculosis drugs in three cities in three geopolitical zones in Nigeria.methods A total of 527 smear-positive sputum samples from Abuja, Ibadan and Nnewi were cultured on BACTEC-MGIT 960. Drug susceptibility tests (DST) for streptomycin, isoniazid, rifampicin and ethambutol were performed on 428 culture-positive samples on BACTEC-MGIT960.results Eight per cent of the specimens cultured were multi-drug-resistant Mycobacterium tuberculosis (MDR-TB) with varying levels of resistance to individual and multiple first-line drugs. MDR was strongly associated with previous treatment: 5% of new and 19% of previously treated patients had MDR-TB (OR 4.1 (95% CI 1.9-8.8), P = 0.001) and with young adult age: 63% of patients with and 38% without MDR-TB were 25-34 years old (P = 0.01). HIV status was documented in 71%. There was no association between MDR-TB and HIV coinfection (P = 0.9) and gender (P > 0.2 for both).conclusions MDR-TB is an emerging problem in Nigeria. Developing good quality drug susceptibility test facilities, routine monitoring of drug susceptibility and improved health systems for the delivery of and adherence to first-and second-line treatment are imperative to solve this problem.keywords multi-drug-resistant tuberculosis, urban cities, first-line TB drugs, newly diagnosed and previously treated TB, HIV

BackgroundThe clinical significance of bacteraemia secondary to non-typhoidal Salmonella (NTS) gastroenteritis in hospitalised adults is uncertain.MethodsAdults admitted to a hospital in Liverpool, UK, with NTS gastroenteritis were identified using hospital discharge data and laboratory records. Patients with known HIV infection were excluded. Risk factors for a complicated or fatal course were determined.ResultsBetween 1982 and 2006 inclusive, 633 adults were identified. Serovars causing infection included Enteritidis (46.6%), Typhimurium (27.6%) and Virchow (4.9%). A blood culture was taken in 364 (57.5%) patients who were generally sicker than those who were not cultured. Bacteraemia was detected in 63 (17.3%) patients who had blood cultures taken (63/633 (10.0%) of all patients). Bacteraemia was more common in those aged ≥ 65 years (p < 0.001) and in those aged < 65 years who had an underlying chronic disease. A complicated course occurred in 91 (25.0%) patients who had had a blood culture taken (148/633 (23.4%) of all patients). Independent factors associated with a complicated or fatal course among the patients investigated with a blood culture were bacteraemia (Adjusted Odds Ratio 5.34, 95% CI 2.86–9.95); new onset confusion or coma (AOR 4.80, 95% CI 1.91–12.07); prolonged symptoms prior to admission (AOR 2.48, 95% CI 1.44–4.27); dehydration (AOR1.90, 95% CI 1.07–3.38); and absence of fever (AOR 0.56, 95% CI 0.32–0.95). The 30 day attributable case fatality for all patients was 1.5%.ConclusionsIn this study secondary bacteraemia, as well as other clinical factors, was independently associated with a complicated or fatal course in non-HIV infected adults admitted to hospital with NTS gastroenteritis.

BSI is a common problem in Cambodian children attending hospital and associated with significant mortality. Further studies are needed to clarify the epidemiology of neonatal sepsis, the contribution of atypical organisms and the epidemiology of pneumococcal disease before the introduction of vaccine.