Characterization of Candida species isolated from the hands of the healthcare workers in the neonatal intensive care unit.

Colonization of health professional hands by potentially pathogenic microorganisms capable to expressing virulence factors, such as Candida spp., is worrisome because of the high contact between patients and professionals. The study aims to evaluate, in vitro, the following virulence factors: hemolytic activity, DNAse expression, biofilm formation, and susceptibility for antifungal agents of Candida species isolated from health professionals hands of a neonatal intensive care unit (NICU). The study includes 50 isolates of Candida spp.: 19 C. parapsilosis sensu stricto, three C. metapsilosis, one C. orthopsilosis, seven C. albicans, six C. famata, five C. lusitaniae, three C. krusei, two C. kefyr, two C. tropicalis, one C. glabrata, and one C. guilliermondii. The hemolytic activity and DNAse were investigated using blood agar and DNAse agar, respectively. Biofilm production was evaluated through XTT sodium salt reduction ability, and the susceptibility of the isolates to antifungals through the microdilution methodology. Forty-nine isolates presented at least one of the three virulence factors investigated. C. albicans showed more intense hemolytic activity. DNAse production was statistically significant between the C. parapsilosis complex and C. albicans, as well as between the C. parapsilosis complex and C. krusei, even as between C. famata and C. albicans, and between C. famata and C. krusei. Forty-three isolates produced biofilm. Seventy-eight percent of the isolates were sensitive to the three antifungals tested. This study demonstrated that Candida isolated from healthcare professionals' hands has virulence.

Quality of human milk expressed in a human milk bank and at home / Qualidade do leite humano ordenhado em banco de leite e no domicílio

Abstract Objective: To evaluate the quality of the human milk expressed at home and at a human milk bank. Methods: This a retrospective, analytical, and observational study, performed by assessing titratable acidity records and the microbiological culture of 100 human milk samples expressed at home and at a human milk bank, in 2014. For the statistical analysis, generalized estimating equations (GEE) and the chi-squared test were used. Results: When comparing the two sample groups, no significant difference was found, with 98% and 94% of the samples being approved among those collected at the milk bank and at home, respectively. No main interaction effect between local and titratable acidity records (p = 0.285) was observed, and there was no statistically significant difference between the expected and observed values for the association between the collection place and the microbiological culture results (p = 0.307). Conclusions: The quality of human milk expressed at home and at the milk bank are in agreement with the recommended standards, confirming that the expression of human milk at home is as safe as expression at the human milk bank, provided that the established hygiene, conservation, storage, and transport standards are followed.

Resumo Objetivo: Avaliar a qualidade do leite humano ordenhado em domicílio e no Banco de Leite Humano. Métodos: Estudo retrospectivo, feito por meio da avaliação dos registros da acidez titulável e dos resultados de cultura microbiológica de 100 amostras de leite humano ordenhado em domicílio e em um Banco de Leite Humano, no ano de 2014. Para análises estatísticas foram utilizadas as Equações de Estimação Generalizadas (Generalized Estimating Equations - GEE) e o teste Qui-quadrado. Resultados: Na comparação dos dois grupos de amostras, não foi detectada diferença significativa, sendo que 98% e 94% das amostras foram aprovadas entre as coletadas no Banco de Leite e em domicílio, respectivamente. Não foi observado efeito principal de interação entre local e grau de acidez titulável (p = 0,285) e não se constatou diferença estatisticamente significante entre os valores observados e esperados para associação entre local de coleta e o resultado da cultura microbiológica (p = 0,307). Conclusões: A qualidade do leite humano ordenhado em domicílio e no Banco de Leite Humano estão de acordo com o padrão preconizado, comprovando que a ordenha de leite humano em domicílio é tão segura quanto a ordenha no Banco de Leite Humano, desde que sejam seguidas as normas de higiene, conservação, armazenamento e transporte estabelecidas.

Quality of human milk expressed in a human milk bank and at home.

OBJECTIVE: To evaluate the quality of the human milk expressed at home and at a human milk bank. METHODS: This a retrospective, analytical, and observational study, performed by assessing titratable acidity records and the microbiological culture of 100 human milk samples expressed at home and at a human milk bank, in 2014. For the statistical analysis, generalized estimating equations (GEE) and the chi-squared test were used. RESULTS: When comparing the two sample groups, no significant difference was found, with 98% and 94% of the samples being approved among those collected at the milk bank and at home, respectively. No main interaction effect between local and titratable acidity records (p=0.285) was observed, and there was no statistically significant difference between the expected and observed values for the association between the collection place and the microbiological culture results (p=0.307). CONCLUSIONS: The quality of human milk expressed at home and at the milk bank are in agreement with the recommended standards, confirming that the expression of human milk at home is as safe as expression at the human milk bank, provided that the established hygiene, conservation, storage, and transport standards are followed.

Fatores de risco para colonização e sepse por Candida albicanse Candida não albicans em neonatos críticos / Risk factors for colonization and sepsis by Candida albicans and non-albicans Candida in critical newborns

O objetivo do estudo foi avaliar as taxas de sepse e de colonização por Candida albicans e Candida não albicans e os fatores de risco relacionados em neonatos críticos. O estudo foi realizado na Unidade de Terapia Intensiva Neonatal (UTIN) do Hospital de Clínicas da Universidade Federal de Uberlândia, com 10 leitos, através de dois tipos de vigilância epidemiológica: “National Nosocomial Infections Surveillance”(NNIS) e laboratorial, no período de Jan/02 a Dez/07. As amostras de Candida isoladas de infecção foram obtidas no Laboratório de Microbiologia do hospital. Os neonatos foram avaliados quanto à colonização nas mucosas oral e intestinal, após 24 horas da internação na unidade e em intervalos semanais até a alta, utilizando-se “swab” estéril, cultivado em “Cromo-agar”. No total, a taxa de sepse foi 13,6%, sendo que as candidemias responderam por 6% das mesmas, todas de natureza hospitalar, associadas com uma letalidade de 33,3%. A taxa de infecção por Candida/ 1.000 pacientes dia foi de 0,96, sendo a maioria (47,4%; 9/19) por Candida albicans. Foram constatados 19 (54,28%) neonatos colonizados, dos quais originaram 35 amostras, 15 de mucosa bucal com 53,3% por Candida albicans e 46,7% Candida não albicans, e 20 de mucosa perianal com 50,0% por Candida albicans. Os fatores de risco associados com a candidemia invasiva (P d” 0,05) foram: uso de CVC e de antibióticos, e para colonização por Candida foram: nutrição parenteral, entubação e uso prévio de antibiótico com um predomínio de cefalosporina de terceira geração (83,3%). Não foi observada relação entre colonização prévia e candidemia invasiva, com o de apenas uma criança com sepse por Candida albicans, colonizada previamente.

The objective of this study was to evaluate the rates of sepsis and colonization by Candida albicans andnon- albicans Candida and the risk factors observed in critical newborns. The study was carried out in a 10-bed Neonatal Intensive Care Unit (NICUs) of the Clinical Hospital of the Federal University of Uberlândia, through two types of epidemiological surveillance: “National Nosocomial Infections Surveillance" (NNIS) and laboratorial in the period of Jan/02 to Dec/07. Samples of Candida isolated from infection were obtained in the Laboratory of Microbiology of the hospital. The newborns were evaluated according to oral and intestinal colonization, after 24 hours of hospitalization in that unit in weekly intervals until discharge. Overall, the rate of sepsis was 13.6%, and the candidemias accounted for 6%, from hospital nature , associated with 33.3% of lethality. The rate of infection by Candida/1,000 patient daily was 0.96, while the majority (47.4%; 9/19) by Candida albicans. Nineteen (54.28%) neonates colonized were observed from them 35 samples were provided, 15 of oral mucosa with 53.3% by Candida albicans, and 46.7% by non-albicans Candida, and 20 of mucous perianal with 50.0% by Candida albicans. The risk factors associated with invasive candidemia (P d” 0.05) were: use of CVC and antibiotics, and colonization by Candida were: parenteral nutrition, intubation and previous use of antibiotic with a predominance of the third generation cephalosporin ( 83.3%). There was no relationship between prior colonization and invasive candidemia, with only one child with sepsis by Candida albicans, previously colonized.

Two outbreaks of mixed etiology associated with central venous catheters inserted by phlebotomy in critical neonates

Staphylococcus aureus and coagulase-negative staphylococci are the main cause of sepsis in Neonatal Intensive Care Unit (NICU). Central venous catheters (CVCs) are an important part of critical neonates' treatment and are associated with sepsis. The aim of this study was to investigate two outbreaks caused by Staphylococcus aureus and Staphylococcus epidermidis associated with CVC inserted by phlebotomy in critical neonates. The surveillance was performed from January 2001 to December 2005 at the Brazilian NICU. The genotypic analysis of oxacillin susceptible S. aureus (OSSA) and oxacillin resistant S. epidermidis (ORSE) was performed based on pulsed-field gel electrophoresis (PFGE). Staphylococcus was the most frequent pathogen (65.8 percent) with highest incidence of CoNS (59.9 percent) followed by S. aureus (40.1 percent). During the five years of surveillance, there were two outbreaks detected, occurred in January-February/02 and August/02 and confirmed by PFGE analysis. The predisposing factors for infection corresponding to both outbreaks were: age <7 days, hospitalization > 7 days, and use of polyethylene CVC through dissection of vein (phlebotomy). This is the first relate of staphylococcal outbreaks associated with CVC inserted by phlebotomy in NICU. PFGE showed polyclonal spread of OSSA during both epidemic and endemic period, and two monoclonal outbreaks of ORSE in the same epidemic period of OSSA.

Molecular surveillance of methicillin-susceptible Staphylococcus aureus at a neonatal intensive care unit in Brazil.

BACKGROUND: We evaluated the relationship among hospital infection and colonization by methicillin-susceptible Staphylococcus aureus (MSSA), clonal spread, and associated risk factors in a neonatal intensive care unit (NICU) of the Uberlândia Federal University-affiliated hospital in Brazil. METHODS: Between February 2004 and June 2005, a longitudinal surveillance study was carried out in an NICU with neonates presenting infections, through both the NNIS system and S aureus punctual colonization prevalence inquests. RESULTS: The overall rate of infection incidence was 23/1000 patient-days. Of all the neonates assessed, 15 were infected and 15 colonized. Sepsis was the most frequent infection, whereas anterior nare was the most isolated site. Antibiotics use, central vascular catheter (CVC), and CVC use more than 7 days and its insertion by phlebotomy were the risk factors for colonization/infection. Molecular analysis showed polyclonal origin (12 genotypes), with predominance of a genotype ("B"), and clonal identity between colonization and infection samples. CONCLUSION: The analysis by means of classical epidemiology and molecular techniques pointed out that methicillin-susceptible Staphylococcus aureus infections were associated with previous colonization by the pathogen, with evidence of horizontal transmission within the unit.

Two outbreaks of mixed etiology associated with central venous catheters inserted by phlebotomy in critical neonates.

Staphylococcus aureus and coagulase-negative staphylococci are the main cause of sepsis in Neonatal Intensive Care Unit (NICU). Central venous catheters (CVCs) are an important part of critical neonates' treatment and are associated with sepsis. The aim of this study was to investigate two outbreaks caused by Staphylococcus aureus and Staphylococcus epidermidis associated with CVC inserted by phlebotomy in critical neonates. The surveillance was performed from January 2001 to December 2005 at the Brazilian NICU. The genotypic analysis of oxacillin susceptible S. aureus (OSSA) and oxacillin resistant S. epidermidis (ORSE) was performed based on pulsed-field gel electrophoresis (PFGE). Staphylococcus was the most frequent pathogen (65.8%) with highest incidence of CoNS (59.9%) followed by S. aureus (40.1%). During the five years of surveillance, there were two outbreaks detected, occurred in January-February/02 and August/02 and confirmed by PFGE analysis. The predisposing factors for infection corresponding to both outbreaks were: age < or =7 days, hospitalization > or =7 days, and use of polyethylene CVC through dissection of vein (phlebotomy). This is the first relate of staphylococcal outbreaks associated with CVC inserted by phlebotomy in NICU. PFGE showed polyclonal spread of OSSA during both epidemic and endemic period, and two monoclonal outbreaks of ORSE in the same epidemic period of OSSA.

Specific IgA and IgG antibodies in paired serum and breast milk samples in human strongyloidiasis.

Strongyloidiasis, caused by the nematode Strongyloides stercoralis, is one of the major worldwide parasitic infections in humans. Breastfeeding may offer a potential protection against this infection. Feces, serum and milk samples were obtained from 90 lactating women from Clinical Hospital of Universidade Federal de Uberlândia, Brazil. The fecal samples were collected for parasitological diagnosis and the serum and milk samples were examined for specific S. stercoralis IgA and IgG antibodies using the indirect fluorescent antibody test (IFAT) and enzyme-linked immunosorbent assay (ELISA). Fecal examination showed that the rate of prevalence of S. stercoralis infection in the lactating women was 4.4%. IFAT manifested a 16.7% positivity rate for specific IgA antibody in serum and a 28.9% rate in milk samples; specific IgG was 41.1% in serum and 25.5% in milk samples. According to ELISA the positivity rate for specific IgA antibody was 21.1% in serum and 42.2% in milk samples; specific IgG was 40% in serum and 18.9% in milk samples. In serum samples, these immunological tests showed a concurrence of 91.1% and 94.4%, respectively, in detecting specific IgA and IgG antibodies. In milk samples, they showed a concurrence of 70% and 78.9%, respectively, in detecting specific IgA and IgG antibodies. There was a statistically significant difference between concordant and discordant results of immunological tests (P<0.0001). IFAT and ELISA highly concurred in their detection of specific S. stercoralis IgA and IgG antibodies in serum and in milk samples reconfirming prior studies that the serological method is a complement to the direct diagnosis of the parasite, and suggesting that immunological methods using milk samples can also be helpful. Furthermore, in endemic areas, infants may acquire antibodies to S. stercoralis from breast milk, possibly, contributing to the enhancement of specific mucosal immunity against this parasite.

Formação de biofilme em amostras de Staphylococcus epidermidis isoladas de sepse relacionada a cateter vascular central em neonatos críticos / Biofilm production in Staphylococcus epidermidis samples isolated fromsepsis associated with central vascular catheter (CVC) in critical neonates

Esta pesquisa teve como objetivo verificar a presença de biofilme em amostras de S. epidermidis isoladas de sangue em casos de sepse relacionados ao uso de CVC em neonatos críticos. O estudo foi realizado na Unidade de Terapia Intensiva Neonatal (UTIN) do Hospital de Clínicas da Universidade Federal de Uberlândia, no período de Jan/02 a Dez/06. Adicionalmente, foi realizado inquéritos de prevalência pontual de colonização por este microrganismo, através da coleta de espécimes da narina e axila com auxílio de swabs, entre Janeiro de 2004 e Junho de 2005. A avaliação de formação de biofilme foi realizada por densidade ótica em placas de poliestireno, após coloração com safranina, utilizando espectrofotômetro (570nm). O teste de suscetibilidadeà oxacilina foi realizado pelo método de diluição de acordo com o “Clinical and Laboratory Standards Institute”.Foram analisadas 109 amostras de S. epidermidis de episódios de sepse, sendo 83 (76,1%) relacionadas ao uso de CVC. Em relação à presença de biofilme, 67/83 (80,7%) das amostras isoladas de neonatos com sepserelacionadas a CVC, apresentaram produção intensa, enquanto que entre as 26 não relacionadas ao uso de CVC apenas seis (23,1%) demonstraram produção de biofilme, assim como, 10 (12,3%) entre as associadas à colonização. Cerca de 73% das amostras foram resistentes à oxacilina (concentração inibitória mínima ³0,5mg/ml), incluindo 56 (83,6%) correspondentes a casos de infecção relacionados à CVC, seis (100%) amostras não relacionadas ao uso de CVC e em quatro (40%) de amostras de colonização. Conclui-se que a maioria das amostras de S. epidermidis isoladas de sepse relacionadas à CVC foram biofilme positivas e resistente à oxacilina

This research aimed to observe the biofilm production by Staphylococcus epidermidis isolated associated with central vascular catheter (CVC) related bacteremia in critical neonates. The study it was carried out in Neonatal Intensive Care Unit of the Hospital das Clínicas da Universidade Federal de Uberlândia, in the period of Jan/02 to Dez/06. Additionally, it was carried out inquiries of prevalence, through the specimen collection of the nostril and armpit with aid of swabs, between Jan/04 and Jun/05. The evaluation of biofilm production was by optical density in polystyrene plates, stained with safranin and measured with ELISA reader at 570nm. The oxacilin susceptibility test was carried out through to the dilution method in accordance with the “Clinical and Laboratory Standards Institute”. It was analyzed 109 S. epidermidis samples, being 83(76.1%) related to the use of CVC. In relation to the biofilm presence, 80.7% of the neonates isolated samples with sepsis related the CVC, had presented intense production, whereas of the 26 unrelated ones to the useof CVC, only six (23.1%) had demonstrated biofilm production as well as 10 (12.3%) of the samples associated to the colonization. About 73% of the samples were oxacilin resistant (minimum inhibitory concentration³0.5mg/ml) . In conclusion, the S. epidermidis samples isolated of CVC related sepsis were biofilm positive and pertaining to oxacilin resistant phenotype

Crescimento de recém-nascido pré-termo / Growth of preterm infants

Acompanharam-se 141 recém-nescidos pré-termo com idade de 26 a 36 semanas ao nascer, adequados para a idade gestacional, do nascimento até a idade pos-menstrual corrigida de 42 semanas. As medidas do peso, comprimento e perimetro cefalico foram obtidas semanalmente. A partir dos valores médios e dos percentuais dessas medidas, foi ajustada uma funçäo polinomial,conseguindo melhor ajuste com o polinômio de 3§ grau.As curvas de crescimento obtidas, quando comparadas com as chamadas curvas de crescimento intra-uterino, mostram que na idade da 40ª a 42ª semanas pos-menstrual as médias ou medianas säo semelhantes. Observou-se uma dinâmica do crescimento dos recém-nascidos pré-termo com caracteristicas de um crescimento de recuperaçäo .