Prevention of nosocomial legionellosis by best water management: comparison of three decontamination methods.

BACKGROUND: Since 2000, the National Health System has adopted international guidelines for assessing Legionella spp. in hospital water systems. The control of water contamination by Legionella spp. is still a matter of research concerning the most effective method in preventing nosocomial infections. AIM: To compare three different decontamination methods by monitoring colony-forming unit count and number of hospital-acquired legionellosis cases. A secondary objective was to evaluate the long-term effects of the preventive measures on the water pipes. METHODS: A protocol was developed for the selection of high-risk sampling sites and for the testing of three disinfection methods over the course of 19 years: hyperchlorination and thermal shock (period A, 2000-2005); copper-silver ionization (period B, 2006-2010); and integration of pre-filtering, filtering, pipe-protecting products, and remote control with chlorine dioxide (ClO2) (period C, 2011-2018). FINDINGS: The use of shock disinfection and hyperchlorination led to a decrease in contamination level immediately after the procedure, but then it rose again to the previous level in two months. Both copper-silver ionization and ClO2 disinfection showed a stable and durable decrease in contamination level. Throughout these three phases, six cases of Legionella spp. occurred during period A, six cases during period B, and three cases during period C. With regard to the damage of water pipes, effective copper-silver levels caused corrosion and calcification in water pipes. CONCLUSION: Both copper-silver ionization and ClO2 properly controlled Legionella spp. contamination. ClO2 significantly reduced the number of positive sites (P < 0.001) without damaging the pipelines.

Genome wide selection in Citrus breeding.

Genome wide selection (GWS) is essential for the genetic improvement of perennial species such as Citrus because of its ability to increase gain per unit time and to enable the efficient selection of characteristics with low heritability. This study assessed GWS efficiency in a population of Citrus and compared it with selection based on phenotypic data. A total of 180 individual trees from a cross between Pera sweet orange (Citrus sinensis Osbeck) and Murcott tangor (Citrus sinensis Osbeck x Citrus reticulata Blanco) were evaluated for 10 characteristics related to fruit quality. The hybrids were genotyped using 5287 DArT_seqTM (diversity arrays technology) molecular markers and their effects on phenotypes were predicted using the random regression - best linear unbiased predictor (rr-BLUP) method. The predictive ability, prediction bias, and accuracy of GWS were estimated to verify its effectiveness for phenotype prediction. The proportion of genetic variance explained by the markers was also computed. The heritability of the traits, as determined by markers, was 16-28%. The predictive ability of these markers ranged from 0.53 to 0.64, and the regression coefficients between predicted and observed phenotypes were close to unity. Over 35% of the genetic variance was accounted for by the markers. Accuracy estimates with GWS were lower than those obtained by phenotypic analysis; however, GWS was superior in terms of genetic gain per unit time. Thus, GWS may be useful for Citrus breeding as it can predict phenotypes early and accurately, and reduce the length of the selection cycle. This study demonstrates the feasibility of genomic selection in Citrus.

Diversity and Genetic Variation among Brevipalpus Populations from Brazil and Mexico.

Brevipalpus phoenicis s.l. is an economically important vector of the Citrus leprosis virus-C (CiLV-C), one of the most severe diseases attacking citrus orchards worldwide. Effective control strategies for this mite should be designed based on basic information including its population structure, and particularly the factors that influence its dynamics. We sampled sweet orange orchards extensively in eight locations in Brazil and 12 in Mexico. Population genetic structure and genetic variation between both countries, among locations and among sampling sites within locations were evaluated by analysing nucleotide sequence data from fragments of the mitochondrial cytochrome oxidase subunit I (COI). In both countries, B. yothersi was the most common species and was found in almost all locations. Individuals from B. papayensis were found in two locations in Brazil. Brevipalpus yothersi populations collected in Brazil were more genetically diverse (14 haplotypes) than Mexican populations (four haplotypes). Although geographical origin had a low but significant effect (ca. 25%) on the population structure, the greatest effect was from the within location comparison (37.02 %). Potential factors driving our results were discussed.

Citrus leprosis virus C Naturally Infecting Commelina benghalensis, a Prevalent Monocot Weed of Citrus Orchards in Brazil.

There are two bacilliform, rhabdo-like viruses that cause citrus leprosis: Citrus leprosis virus C (CiLV-C), which accumulates in the cytoplasm of infected cells, and Citrus leprosis virus nuclear type (CiLV-N), which accumulates in their nucleus. The first one, the prototype of the new genus Cilevirus, is prevalent and occurs in several countries of the American continent, from Argentina to Mexico (1). The second type, still a tentative member of the Rhabdoviridae family, is of rare occurrence, with a few reports in Brazil and one in Panama (1). Leprosis is particularly important to the Brazilian citrus industry because of the 60 to 80 million dollars spent yearly for the control of Brevipalpus phoenicis (Geijskes, 1939) (Acari: Tenuipalpidae), the vector of the virus (1). For decades, the disease was considered unique to citrus plants; however, greenhouse experiments conducted in the 1990s demonstrated the mechanical transmission of CiLV-C to noncitrus plants (1). Years later, researchers were able to transmit the virus to nonrutaceous hosts using viruliferous mites (1,4). Recently, León et al. (2) reported the occurrence of the first noncitrus plant naturally infected by CiLV-C, the rutaceous Swinglea glutinosa Blanco (Merr.). Tropical spiderworts (Commelina benghalensis L.; Commelinaceae) are monocot weeds commonly found in citrus orchards in Brazil. In a survey conducted in orchards with high incidences of leprosis in the municipalities of Borborema and São José do Rio Preto, State of Sao Paulo, Brazil, tropical spiderworts were found exhibiting necrotic spots with a yellow halo in green leaves and green spots with necrotic center in senescent leaves. Since these symptoms are similar to those caused by CiLV-C in citrus, symptomatic plants were collected and subjected to transmission electron microscopy analyses and reverse transcription-PCR using primers that specifically amplify a region within the putative movement protein gene of the virus (3). Bacilliform virus particles and typical inclusions were seen in the lesions. Bands of the expected 344 bp size were seen in agarose gels of symptomatic samples only. The analysis of the consensus sequence showed 100% identity with CiLV-C sequence available in the GenBank (Accession No. YP_654542.1). Experimental transmission of CiLV-C by B. phoenicis reproduced the lesions in inoculated tropical spiderwort. Also, the virus could be easily transmitted back from C. benghalensis to sweet orange plants. Our data show that this widespread weed is the first monocot as a natural host for CiLV-C. Since tropical spiderwort is a host for B. phoenicis and symptomatic plants were found in two municipalities 130 km apart from each other, it is possible that this weed may serve as reservoir for the virus and play a relevant role in the spread of the disease in the field, but this issue still needs to be addressed in further experiments. References: (1) M. A. Bastianel et al. Plant Dis. 94:284, 2010. (2) M. G. A. Leon et al. Plant Dis. 90:682, 2008. (3) E. C. Locali-Fabris et al. Plant Dis. 87:1317. (4) M. A. Nunes et al. Plant Dis. Online publication. doi:10.1094/PDIS-06-11-0538, 2011.

Detection of Brevipalpus-transmitted viruses in their mite vectors by RT-PCR.

The diagnosis of plant diseases caused by Brevipalpus-transmitted viruses (BrTVs) has been done through the analyses of symptoms, transmission electron microscopy, and RT-PCR of infected plant tissues. Here, we report the detection of Citrus leprosis virus C, Orchid fleck virus, Clerodendrum chlorotic spot virus and Solanum violaefolium ringspot virus in their viruliferous vectors Brevipalpus spp. using specific primer pairs for each of the viruses. The efficiency of virus transmission by Brevipalpus mites is low, so the detection of these pathogens in their vectors could constitute an important tool for studies involving virus-vector relationships, transmission, and monitoring the pathogen prior to the appearance of symptoms in the field.

An evaluation of the completeness of reporting of childhood tuberculosis.

The sensitivity of the Enhanced Tuberculosis Surveillance (ETS) scheme for monitoring tuberculosis in children is unknown. We used the British Paediatric Surveillance Unit (BPSU) reporting scheme to conduct a prospective observational study of tuberculosis in children aged <16 yrs in the UK. Reported cases were then matched with records from the ETS database. A total of 320 cases were reported to the BPSU between January and December 2004. We estimated that there were 557 paediatric cases in England, Wales and Northern Ireland in 2004: 222 (40%) cases reported to both BPSU and ETS, 98 (18%) reported to BPSU but not ETS and 237 (42%) reported to ETS but not BPSU. Children aged <5 yrs were significantly less likely to be reported to ETS compared with older children (p<0.01). There is substantial under-reporting of childhood tuberculosis, especially of children aged <5 yrs. ETS provides a representative picture of the demographics but may miss approximately 20% of cases. This should be taken into account when planning training and resource requirements for tuberculosis. Increased efforts are needed to ensure that all paediatric cases are reported to ETS.

Tuberculosis in the United Kingdom and Republic of Ireland.

AIMS: To describe the clinical features, diagnosis and management of children with tuberculosis in the United Kingdom and Republic of Ireland. METHODS: Cases of culture-confirmed and clinically diagnosed tuberculosis were reported to the British Paediatric Surveillance Unit from December 2003 to January 2005. RESULTS: 385 eligible cases were reported. Pulmonary disease was present in 154 (40%) children. Just over half (197, 51%) of children presented clinically and most of the remainder (166, 43%) at contact tracing. A probable source case was identified for 73/197 (36%) of the children presenting clinically. The majority (253, 66%) of children had a microbiological and/or histological investigation, and culture results were available for 240 (62%), of whom 102 (26%) were culture positive. Drug resistance was reported in 15 (0.4%) cases. 44% (128/292) of non-white children did not receive the recommended quadruple drug therapy. Seven children died. Only 57% (217) of children were managed by a paediatric subspecialist in respiratory or infectious diseases or a general paediatrician with a special interest in one of these areas. Fewer than five cases were reported from 119/143 (83%) respondents and 72 of 96 (75%) centres. CONCLUSIONS: Many paediatricians and centres see few children with tuberculosis. This may affect adherence to national guidelines. Managed clinical networks for children with tuberculosis may improve management and should be the standard of care.

Association of rs2200733 at 4q25 with atrial flutter/fibrillation diseases in an Italian population.

BACKGROUND: Atrial fibrillation (AF) and atrial flutter (AFL) are common cardiac conduction disorders affecting many people. Recent studies on sporadic cases of AF/AFL showed a significant association of the single nucleotide polymorphism rs2200733T with the disease, suggesting a genetic factor in the development of the disease. OBJECTIVES: To determine the association of rs2200733 with AF/AFL derived from an Italian population sample. SUBJECTS: 78 patients with AF/AFL and 348 controls took part in the study. DESIGN: Genetic case-control study. RESULTS: The results indicate that there is a positive, significant association between the rs2200733 T allele and patients with AF/AFL of Italian origin (allelic p<0.001 with OR = 2.17). CONCLUSION: These results derived from a sample of the Italian population agree with previously reported findings from an Icelandic study, which also found that the minor allele rs2200733 was associated with AF/AFL disease.

A survey of tuberculosis services in the UK.

AIM: To determine the provision of services for children with tuberculosis (TB) living in the UK. METHOD: A postal questionnaire was sent to the most appropriate paediatrician and adult physician in every acute hospital trust in the UK. Information was sought on inpatient and outpatient services for children with TB and for children in contact with TB. RESULTS: Responses were received from 323 individuals in 199 of the 205 trusts approached. The median number of children with TB seen per year at each trust was 1.5 (range 0-30). Inpatients were nearly all admitted to paediatric wards (197 (99%) trusts). In 141 trusts (71%) they were looked after solely by paediatricians or jointly by paediatricians and physicians (47 trusts, 24%). 132 (66%) trusts stated there was a named consultant for children with TB. Negative pressure isolation rooms were reported to be available for children in 42 trusts (21%). As outpatients, children with TB were seen in paediatric clinics in 163 (82%) trusts. Only 10 (5%) trusts had designated family TB clinics. Children in contact with TB were managed by paediatricians in 81 (38%) trusts, by physicians in 67 (34%) trusts and jointly in 51 (26%) trusts. 161 (81%) trusts had access to a TB nurse and directly observed therapy (DOTS) was available in 116 (58%) trusts. CONCLUSIONS: Many paediatricians see few children with TB, but most children with TB are looked after by general paediatricians alone. The survey supports national recommendations to develop family clinics and clinical service networks for children with TB, which may improve the care of these children.

Serum leptin concentrations and fat redistribution in HIV-1-infected children on highly active antiretroviral therapy.

OBJECTIVES: HIV-related lipodystrophy is a syndrome of adipose tissue redistribution, dyslipidaemia and insulin resistance. Combination antiretroviral therapy (CART) is a key risk factor. We hypothesized that fat redistribution in HIV-infected children is related to altered endocrine function of adipose tissue, namely leptin secretion. METHODS: Serum leptin and fat redistribution were measured in 104 HIV-infected children in a prospective observational study from 2003 to 2004. Fat redistribution was defined by clinical observation. Body fatness was estimated using body mass index and four skinfold measurements. Serum leptin was determined using an enzyme-linked immunosorbent assay (Quantikine; R&D Systems, Abingdon, UK). Linear analogue models were used to adjust the leptin concentration for body fatness. RESULTS: There was no significant difference in serum leptin among children treated with protease inhibitors (PIs), children on non-PI CART and children not treated with CART (P>0.05). When leptin concentrations were adjusted for body fatness, there was again no difference among PI-treated, non-PI-treated and untreated children. Categorization of CART exposure as never, current or past did not change these results. CONCLUSIONS: There is no evidence that leptin plays any role in lipodystrophy other than reflecting body fatness.

Determination of cadmium and lead in vegetables by stripping chronopotentiometry.

A method for the determination of cadmium and lead in vegetables by stripping chronopotentiometric analysis, after digestion of the sample with concentrated sulphuric acid and dry-ashing, is described. Metal ions were concentrated as their amalgams on a glassy carbon-working electrode previously coated with a thin mercury film and then stripped by a suitable oxidant. Potential and time data were digitally derived and E was plotted versus dt/dE(-1), thus increasing both the sensitivity of the method and the resolution of the analysis. Quantitative analysis was carried out by the method of standard addition; good linearity was obtained in the range of examined concentrations, as was shown by the determination coefficients, which were 0.998 (n = 4) for cadmium and 0.993 (n = 4) for lead. Recoveries of 85-100% for cadmium and of 84-97% for lead were obtained from a sample spiked at different levels. Accuracy was demonstrated by analysis of a matching reference sample of cabbage. The detection limits were 1.8 ng g(-1) of wet mass for cadmium and 5.1 ng g(-1) of wet mass for lead. The relative standard deviations (mean of nine determinations), evaluated on a real sample, were 6.7 and 6.2%, respectively. Results obtained on 10 different commercial samples of pepper (Capsicum annuum), and egg plant (Solanum melongena) were not significantly different from those obtained by graphite furnace atomic absorption spectrophotometry technique. The average content was in the range 3.1-18.6 ng g(-1) for cadmium and 38.2-64.3 ng g(-1) for lead.

HIV evolution: CTL escape mutation and reversion after transmission.

Within-patient HIV evolution reflects the strong selection pressure driving viral escape from cytotoxic T-lymphocyte (CTL) recognition. Whether this intrapatient accumulation of escape mutations translates into HIV evolution at the population level has not been evaluated. We studied over 300 patients drawn from the B- and C-clade epidemics, focusing on human leukocyte antigen (HLA) alleles HLA-B57 and HLA-B5801, which are associated with long-term HIV control and are therefore likely to exert strong selection pressure on the virus. The CTL response dominating acute infection in HLA-B57/5801-positive subjects drove positive selection of an escape mutation that reverted to wild-type after transmission to HLA-B57/5801-negative individuals. A second escape mutation within the epitope, by contrast, was maintained after transmission. These data show that the process of accumulation of escape mutations within HIV is not inevitable. Complex epitope- and residue-specific selection forces, including CTL-mediated positive selection pressure and virus-mediated purifying selection, operate in tandem to shape HIV evolution at the population level.

Decline in mortality, AIDS, and hospital admissions in perinatally HIV-1 infected children in the United Kingdom and Ireland.

OBJECTIVE: To describe changes in demographic factors, disease progression, hospital admissions, and use of antiretroviral therapy in children with HIV. DESIGN: Active surveillance through the national study of HIV in pregnancy and childhood (NSHPC) and additional data from a subset of children in the collaborative HIV paediatric study (CHIPS). SETTING: United Kingdom and Ireland. PARTICIPANTS: 944 children with perinatally acquired HIV-1 under clinical care. MAIN OUTCOME MEASURES: Changes over time in progression to AIDS and death, hospital admission rates, and use of antiretroviral therapy. RESULTS: 944 children with perinatally acquired HIV were reported in the United Kingdom and Ireland by October 2002; 628 (67%) were black African, 205 (22%) were aged > or = 10 years at last follow up, 193 (20%) are known to have died. The proportion of children presenting who were born abroad increased from 20% in 1994-5 to 60% during 2000-2. Mortality was stable before 1997 at 9.3 per 100 child years at risk but fell to 2.0 in 2001-2 (trend P < 0.001). Progression to AIDS also declined (P < 0.001). From 1997 onwards the proportion of children on three or four drug antiretroviral therapy increased. Hospital admission rates declined by 80%, but with more children in follow up the absolute number of admissions fell by only 26%. CONCLUSION: In children with HIV infection, mortality, AIDS, and hospital admission rates have declined substantially since the introduction of three or four drug antiretroviral therapy in 1997. As infected children in the United Kingdom and Ireland are living longer, there is an increasing need to address their medical, social, and psychological needs as they enter adolescence and adult life.

Retrospective screening for hepatitis C in a tertiary paediatric referral centre.

The true prevalence of hepatitis C virus in children in the UK is not known and targeted screening is not standard practice despite an anticipated rise in new cases due to vertical transmission. An extension of the Department of Health's 'look-back' exercise was undertaken in order to determine the prevalence of hepatitis C virus in high-risk patient groups who were transfused with blood and/or blood products before 1991. Five hundred and ninety-five patients transfused between 1971-91 were traced and offered counselling and testing. Blood samples from 405 were analysed for the presence of HCV antibodies and/or HCV RNA by RT-PCR and eight patients were found to be positive. The HCV seroprevalence rate in this cohort was 1.97% and the HCV genome detection rate was 1.72%. In view of the long-term complications from this infection and the availability of potentially effective anti-viral agents, we feel that targeted screening is of value in this setting.

Heterogeneity in the granulomatous response to mycobacterial infection in patients with defined genetic mutations in the interleukin 12-dependent interferon-gamma production pathway.

Patients with genetic lesions in the Type-1 cytokine/cytokine receptor pathway exhibit a selective susceptibility to severe infections with poorly pathogenic mycobacteria and non-typhi salmonella spp. These experiments of nature demonstrate that IL-12-dependent IFNgamma production is critical for granuloma formation and therefore host immunity against such pathogens. The essential role of granuloma formation for protective immunity to these organisms is emphasized by the differing granuloma forming capabilities and resultant clinical sequelae observed in these patients which seems to reflect their ability to produce or respond to IFNgamma (Fig. 9). At one pole of this spectrum, represented by the complete IFNgammaR1/2 deficient patients, there is a complete absence of mature granuloma formation, whereas with the less severe mutations (i.e. partial IFNgammaR1/2, complete IL-12p40 and complete IL-12Rbeta1 deficiency), granuloma formation is very heterogenous with wide variations in composition being observed. This suggests that in the latter individuals, who produce partial but suboptimal IFNgamma responses, other influences, including pathogen virulence and host genotype may also affect the type and scale of the cellular response elicited.

Interferon-gamma receptor deficiency mimicking Langerhans' cell histiocytosis.

Two patients who were initially given a diagnosis of Langerhans' cell histiocytosis on the basis of the clinical, radiologic, and biopsy findings had mycobacterial infection subsequently identified. The correct diagnosis of dominant partial interferon-gamma receptor deficiency was established.

Discitis in young children.

Discitis is uncommon in children and presents in different ways at different ages. It is most difficult to diagnose in the uncommunicative toddler of one to three years of age. We present 11 consecutive cases. The non-specific clinical features included refusal to walk (63%), back pain (27%), inability to flex the lower back (50%) and a loss of lumbar lordosis (40%). Laboratory tests were unhelpful and cultures of blood and disc tissue were negative. MRI reduces the diagnostic delay and may help to avoid the requirement for a biopsy. In 75% of cases it demonstrated a paravertebral inflammatory mass, which helped to determine the duration of the oral therapy given after initial intravenous antibiotics. At a mean follow-up of 21 months (10 to 40), all the spines were mobile and the patients free from pain. Radiological fusion occurred in 20% and was predictable after two years. At follow-up, MRI showed variable appearances: changes in the vertebral body usually resolved at 24 months and recovery of the disc was seen after 34 months.