Orientador: Renato Passini Junior / Dissertação (mestrado) - Universidade Estadual de Campinas, Faculdade de Ciencias Medicas / Made available in DSpace on 2018-11-27T10:57:27Z (GMT). No. of bitstreams: 1 Pinto_GiulianeJesusLajos_M.pdf: 245675 bytes, checksum: 71fd93a1854439989a54c4faa89da2dc (MD5) Previous issue date: 2005 / Resumo: Objetivo: estudar a colonização bacteriana endocervical em gestantes com trabalho de parto prematuro e/ou ruptura prematura de membranas (termo e pré-termo). Método: 212 gestantes com trabalho de parto prematuro (TPP) e/ou ruptura prematura de membranas (RPM), internadas no Hospital Estadual Sumaré (Unicamp), foram avaliadas no período de julho de 2002 a janeiro de 2004. Na admissão hospitalar foram coletadas duas amostras do conteúdo endocervical, realizadas bacterioscopia e cultura em meios ágar-sangue ou ágar-chocolate. Foram analisadas associações da colonização endocervical com infecção de trato urinário materno, corioamnionite, uso de antibióticos, dados de parto, sofrimento fetal, prematuridade, infecção e óbito neonatais. Resultados: entre as mulheres estudadas, 74 (35%) tinham TPP e 138 (65%), RPM. A prevalência de colonização endocervical foi de 14,2% (IC=9,5%-18,9%), com resultados similares em TPP e RPM. Na população estudada, o microorganismo mais encontrado foi o estreptococo do grupo B (EGB) (9,4%), sendo também isolados Candida sp (5 casos), Streptococcus sp (2 casos), Streptococcus pneumoniae, Escherichia coli e Enterococcus sp (1 caso de cada). Das bacterioscopias analisadas, os achados mais freqüentes foram baixa prevalência de bacilos de Dodërlein e elevado número de leucócitos. Em mulheres colonizadas houve maior prevalência de infecção de trato urinário (23,8% versus 5,4%; p<0,01), infecção neonatal (25,0% versus 7,3%; p<0,01) e óbito neonatal (dois casos entre as colonizadas; p<0,02), quando comparadas às não-colonizadas. Conclusões: observou-se alta prevalência de colonização endocervical, sem a utilização de meios de cultura seletivos. O EGB foi o principal microorganismo isolado, reforçando a necessidade de triagem deste agente durante a gestação e nas situações de risco estudadas. Um terço das culturas positivas ocorreram por outros agentes. Estudos complementares são necessários para esclarecer a importância destes achados bacteriológicos no canal endocervical e sua associação com complicações gestacionais, sepse e mortalidade neonatais / Abstract: Objective: to study cervical colonization in women with preterm labor and/or premature rupture of membranes. Method: 212 pregnant women with preterm labor and/or premature rupture of membranes (PROM), admitted at Hospital Estadual Sumaré, during the period between July 2002 and January 2004, were studied. Two cervical samples from each woman were collected and bacterioscopy and culture in blood-agar or chocolate-agar plates were performed. Association of cervical microorganisms and urinary infection, chorioamnionitis, antibiotics use, prematurity, neonatal infection and neonatal death were evaluated. Results: the population evaluated consisted of 74 women with preterm labor (35%) and 138 women with PROM (preterm and term). The prevalence of cervical colonization was 14.2% (CI=9.5-18.9%), with similar results in preterm labor or PROM. Group B streptococcus was the most prevalent organism in this population (9.4%). Other organisms isolated were Candida sp, Streptococcus sp, Streptococcus pneumoniae, Escherichia coli and Enterococcus sp. The most common findings of bacterioscopy were a reduced number of lactobacilli and a great number of leukocytes. Endocervical colonization was associated with a higher occurrence of urinary tract infection (23.8% versus 5.4%; p<0.01), early-onset of neonatal infection (25.0% versus 7.3%; p<0.01) and neonatal mortality (2 cases in colonizated women; p<0.02) when compared with a negative culture of endocervical mucus. Conclusions: this study showed high prevalence of endocervical colonization despite of the use of a nonselective culture media. The main microorganism isolated was Group B streptococcus but other organisms were present in one third of studied population. More studies are needed to evaluate the influence of endocervical colonization in obstetrical outcome and in neonatal sepsis and mortality / Mestrado / Tocoginecologia / Mestre em Tocoginecologia

[Truncated abstract] Fetal inflammatory response syndrome (FIRS) has only recently been recognised as an important cause of spontaneous preterm delivery (PTD). In addition, it has been associated with a number of other short-term and long-term adverse neonatal outcomes, including early onset neonatal sepsis, necrotising enterocolitis, periventricular leucomalacia, cerebral palsy, and bronchopulmonary dysplasia, although the causal mechanisms are unclear. The hallmark of FIRS is histologic chorioamnionitis (HCA). Mothers with HCA are often asymptomatic and it remains unclear whether elevated maternal inflammatory markers, such as C-reactive protein (CRP) and procalcitonin (PCT), are predictive of preterm birth. Furthermore neonatal inflammatory markers such as CRP, PCT, white cell count (WCC) and absolute neutrophil count (ANC), are commonly used in clinical practice to diagnose infection in the neonatal period. Although both intrauterine inflammation and FIRS may have effects on inflammatory markers for up to 10 days following delivery, the extent to which intrauterine infection and FIRS confound these diagnostic surrogates of neonatal infection is unknown. This work addressed the hypothesis that HCA is associated with inflammatory changes that may be detected in the: (a) maternal circulation at the time of delivery, (b) umbilical cord blood at delivery and (c) post-natal circulation within the first 48 hours of life. The primary aim of this study was to investigate the relationship between the presence of HCA and maternal inflammatory markers (serum CRP and PCT on the day of delivery) as well as neonatal inflammatory markers (haematological parameters, CRP and PCT up to 48 hours following delivery). ... Cord platelet counts were likely affected by platelet activation. For both intra-rater and inter-rater reproducibility, the corrected WCC, ANC and NRBC were shown to be reliable with an ICC of >0.90 for all comparisons. However, I:T ratio was poorly reproducible. HCA appears to be a minor inflammatory insult for the mother. In the majority of cases it is asymptomatic and results in minor increases in PCT and CRP levels on the day of delivery. Conversely HCA results in significant inflammatory changes in the newborn that can be seen in the cord blood. Sensitive markers of inflammation in the cord blood are significantly higher in affected infants (CRP and PCT), while less sensitive markers, such as WCC and ANC are not significantly different. This study has shown that fetal inflammation has sustained effects on CRP and haematological parameters in early neonatal life; CRP, WCC and ANC are significantly higher in newborns exposed to HCA, peaking 24 hours following delivery. These effects may confound the interpretation of common diagnostic tests for early onset neonatal sepsis. Conclusion: HCA results in mild elevations in CRP and PCT in the cord blood. Over the subsequent 24 hours CRP, WCC and ANC increase significantly in these neonates. Intrauterine exposure to HCA may influence surrogate diagnostic markers for early onset sepsis in newborn infants. Future research to investigate novel diagnostic markers, such as CD64 and soluble triggering receptor expressed on myeloid cells (TREM-1), or enhanced microbiological molecular diagnosis, will help distinguish true invasive infection from HCA-driven inflammation in the newborn infant.

[Truncated abstract] Pneumococcal exposure is high and life-long in developing countries including Papua New Guinea (PNG), with children under 2 years of age being at most risk for early upper respiratory tract pneumococcal carriage and infection. Deaths from pneumococcal diseases such as pneumonia and meningitis are common and likely the result of an absence of vaccination programmes. The need for effective and affordable pneumococcal vaccines has led to the testing of protein antigens including pneumolysin (Ply) and pneumococcal surface protein A (PspA) as novel vaccine antigens. Little is known on the immune responses to these proteins in humans, particularly in high-risk populations where such vaccines will be of most benefit. In this study, we examined the roles of naturally acquired antibody and cellular immune responses in mothers and newborns to Ply and PspA family 1 (PspA1) and family 2 (PspA2) in protection against or risk for early carriage in a high-risk PNG population. Antibodies to Ply, PspA1 and PspA2 were measured in plasmas of 241 mothers and 115 newborns (cords) from PNG, and 50 Australian mothers using an enzyme-linked immunosorbent assay (ELISA). Pernasal swabs were collected from PNG mothers at the time of delivery, one month post-partum, and weekly within the first month of life from their newborns to determine pneumococcal carriage. Cellular immune responses to Ply, PspA1 and PspA2, the TLR2/TLR4 ligands, LTA and LPS and to PHA were measured in cord blood mononuclear cells (CBMC) of 84 PNG versus 33 Australian newborns. Innate and T-cell cytokine responses in the PNG newborns were then analysed to determine their effect on infant pneumococcal carriage. ... No protective effect against infant pneumococcal carriage was observed with maternal and cord IgG levels for all antigens. Maternal carriage at time of delivery increased the risk for infant pneumococcal carriage in the first month of life (HR: 1.93, 95% CI 1.36 – 2.73, p = 0.001) with 70% of infants being colonised. Papua New Guinean newborns produced higher innate IL-10 and IFN-¿ (p = 0.003) and TNF-a (p < 0.001) to Ply compared to Australian newborns with no significant differences observed for IL-6 or IL-12. IFN-¿ responses to LPS and LTA (p = 0.005 and p < 0.001) were higher in PNG than Australian newborns, while IL-6, IL-10 (p < 0.001) and TNF-a (p = 0.002) to LPS with LTA-induced IL-6 and IL-10 (p < 0.001) were higher in Australian newborns. T-cell IL-5, IL-10, IL-13, IFN-¿, IL-6 and TNF-a response levels to PspA and PHA stimulation were significantly high in PNG newborns. No differences were observed for cytokine responses to Ply and PspA between PNG infant pneumococci carriers and non-carriers. Papua New Guinean infants are colonised by pneumococci very early in life and this may be influenced by high maternal carriage rates. PspA- and Ply-IgG levels are high in PNG mothers and undergo cross placental transfer but do not appear to be protective against early pneumococcal carriage. In PNG newborns, PspA elicits T-cell responses, while Ply drives more innate cellular responses, neither were demonstrated to have a protective effect against early carriage though further work is required to better define these and their relation to immune development in early childhood.

BACKGROUND AND OBJECTIVES: Streptococcus agalactiae (Group B Streptococcus, GBS) is the leading cause of neonatal infections and deaths in human. It can also cause infections in pregnant women and non-pregnant adults. Penicillin and ampicillin are antibiotics of choice for the treatment of GBS infections. Erythromycin and clindamycin are used as alternative therapy in penicillin allergic patients, however resistance to these agents has been increasingly observed. This present study was undertaken to determine the colonization rate of GBS, susceptibility profile and the mechanism of antibiotic resistance in pregnant women and their babies at Dr. George Mukhari Academic Hospital in Pretoria. METHODS: Rectal and vaginal swabs were collected from pregnant women; ear and umbilical swabs from newborns over an 11 month period. Samples were cultured on selective media (CNA agar and Todd-Hewitt broth) and GBS positively identified using morphological and biochemical tests including Gram staining, hemolytic activity, catalase test, bile esculin, CAMP test and Latex agglutination test. The susceptibility testing was done using the Kirby-Bauer and E-test methods. The D-test method was used to determine the inducible clindamycin resistance. Multiplex PCR with were used to detect different genes coding for resistance. RESULTS: Out of the 413 patients evaluated, 128 (30.9%) were positive with GBS. All isolates were sensitive to penicillin and ampicillin. Erythromycin and clindamycin resistance was 21.1% and 17.2% respectively; of which 69% harbouring constitutive MLBB, 17.4% inducible MLSB. The alteration of ribosomal target encoded by ermB genes was the commonest mechanism of resistance observed in 55% of isolates, 38% of isolates had both ermB and linB genes and efflux pump mediated by mefA genes was detected in one of isolates. Conclusion: This study reaffirms the appropriateness of penicillin as the antibiotic of choice for treating GBS infection. However it raises the challenges of resistance to the macrolides and lincosamides. More GBS treatment options for penicillin allergic patients need to be researched. / Health Studies / M.Sc. (Life Sciences (Microbiology))

BACKGROUND AND OBJECTIVES: Streptococcus agalactiae (Group B Streptococcus, GBS) is the leading cause of neonatal infections and deaths in human. It can also cause infections in pregnant women and non-pregnant adults. Penicillin and ampicillin are antibiotics of choice for the treatment of GBS infections. Erythromycin and clindamycin are used as alternative therapy in penicillin allergic patients, however resistance to these agents has been increasingly observed. This present study was undertaken to determine the colonization rate of GBS, susceptibility profile and the mechanism of antibiotic resistance in pregnant women and their babies at Dr. George Mukhari Academic Hospital in Pretoria. METHODS: Rectal and vaginal swabs were collected from pregnant women; ear and umbilical swabs from newborns over an 11 month period. Samples were cultured on selective media (CNA agar and Todd-Hewitt broth) and GBS positively identified using morphological and biochemical tests including Gram staining, hemolytic activity, catalase test, bile esculin, CAMP test and Latex agglutination test. The susceptibility testing was done using the Kirby-Bauer and E-test methods. The D-test method was used to determine the inducible clindamycin resistance. Multiplex PCR with were used to detect different genes coding for resistance. RESULTS: Out of the 413 patients evaluated, 128 (30.9%) were positive with GBS. All isolates were sensitive to penicillin and ampicillin. Erythromycin and clindamycin resistance was 21.1% and 17.2% respectively; of which 69% harbouring constitutive MLBB, 17.4% inducible MLSB. The alteration of ribosomal target encoded by ermB genes was the commonest mechanism of resistance observed in 55% of isolates, 38% of isolates had both ermB and linB genes and efflux pump mediated by mefA genes was detected in one of isolates. Conclusion: This study reaffirms the appropriateness of penicillin as the antibiotic of choice for treating GBS infection. However it raises the challenges of resistance to the macrolides and lincosamides. More GBS treatment options for penicillin allergic patients need to be researched. / Health Studies / M. Sc. (Life Sciences (Microbiology))

Background: Group B streptococcus (GBS) is regarded as one of the most important causes of maternal and neonatal morbidity and mortality in many parts of the world. GBS recto-vaginal colonization is important in the health of a mother and her neonate, especially in developing countries. Maternal vaginal colonization with GBS at the time of delivery can cause vertical transmission to the neonate. Multilocus sequence typing (MLST) is a technique used to characterize microbial isolates by means of sequencing internal fragments of housekeeping genes and has the advantage of reproducibility and has been shown to correlate with the other typing techniques and thus has emerged as the standard for delineating the clonal population of GBS. The study aimed to investigate the epidemiology of GBS colonization among pregnant women and their neonates, and to characterize the isolates by multilocus sequence typing technique at Dr George Mukhari Academic Hospital, Pretoria. Methodology: A total of 413 pregnant women who visited the antenatal clinic were recruited and screened. Participants were interviewed using a questionnaire to gather demographic and other relevant information such as history of current pregnancy, previous miscarriages and still births. Samples from maternal rectum and vagina as well as neonate ear and umbilical cord were taken for culture using colistin and nalidixic acid (CNA) blood agar and incubated for 24-48 hours. If negative after 48 hours, Todd-Hewitt broth was subcultured after 18-48 hours onto sheep blood agar. Multilocus sequence typing (MLST) was used to characterize seven group B streptococcus isolates collected at Dr George Mukhari academic hospital. Fragments of seven housekeeping genes were amplified by polymerase chain reaction (PCR) for each strain and sequenced. CLC bio software (Inqaba biotech, South Africa; Pretoria) was used to analyse sequenced loci and UPGMA dendrogram was constructed. Results: The colonization rate for GBS in pregnant women and their neonates was 30.9% and 0%, respectively. A higher proportion of GBS were isolated from the rectum (37.9%) as compared to the vagina (20.6%). Most socio-economic, demographic and obstetric factors analysed were not significantly associated with.GBS colonization. On 128 positive samples, the results of Todd-Hewitt enrichment broth and direct plating method using CNA were compared. A total of 45.3% of colonised were positive on direct selective agar (CNA); an additional 54.7% samples were recovered from Todd-Hewitt broth. Three genes (adhP, glnA and tkt) were sequenced successfully for six samples (1, 2. 4,6,12 and 65). The UPGMA tree with 1000 bootstrap showing the relationship between six samples was drawn.Conclusion: This study revealed that pregnant women of all ages are at risk of group B streptococcus colonization. Group B streptococcus was common among pregnant women at Dr George Mukhari Academic Hospital. No socio-economic risk factor was associated with group B streptococcus colonization. Results confirm that the combination of Todd-Hewitt broth and CNA agar plate is a time saving and sensitive method. The allelic profile, characteristics such as G+C (guanine+cytosine) content and dN/dS ratio were not analysed because of the smaller sample size used in this study, which shows that the MLST method was unsuccessful in this study. The UPGMA tree based on differences in consensus of the isolates showed that all group B streptococcus isolates are clustered and descend from a single node. / Life & Consumer Sciences / Life Sciences / M.Sc. (Life Sciences)

Background: Group B streptococcus (GBS) is regarded as one of the most important causes of maternal and neonatal morbidity and mortality in many parts of the world. GBS recto-vaginal colonization is important in the health of a mother and her neonate, especially in developing countries. Maternal vaginal colonization with GBS at the time of delivery can cause vertical transmission to the neonate. Multilocus sequence typing (MLST) is a technique used to characterize microbial isolates by means of sequencing internal fragments of housekeeping genes and has the advantage of reproducibility and has been shown to correlate with the other typing techniques and thus has emerged as the standard for delineating the clonal population of GBS. The study aimed to investigate the epidemiology of GBS colonization among pregnant women and their neonates, and to characterize the isolates by multilocus sequence typing technique at Dr George Mukhari Academic Hospital, Pretoria. Methodology: A total of 413 pregnant women who visited the antenatal clinic were recruited and screened. Participants were interviewed using a questionnaire to gather demographic and other relevant information such as history of current pregnancy, previous miscarriages and still births. Samples from maternal rectum and vagina as well as neonate ear and umbilical cord were taken for culture using colistin and nalidixic acid (CNA) blood agar and incubated for 24-48 hours. If negative after 48 hours, Todd-Hewitt broth was subcultured after 18-48 hours onto sheep blood agar. Multilocus sequence typing (MLST) was used to characterize seven group B streptococcus isolates collected at Dr George Mukhari academic hospital. Fragments of seven housekeeping genes were amplified by polymerase chain reaction (PCR) for each strain and sequenced. CLC bio software (Inqaba biotech, South Africa; Pretoria) was used to analyse sequenced loci and UPGMA dendrogram was constructed. Results: The colonization rate for GBS in pregnant women and their neonates was 30.9% and 0%, respectively. A higher proportion of GBS were isolated from the rectum (37.9%) as compared to the vagina (20.6%). Most socio-economic, demographic and obstetric factors analysed were not significantly associated with.GBS colonization. On 128 positive samples, the results of Todd-Hewitt enrichment broth and direct plating method using CNA were compared. A total of 45.3% of colonised were positive on direct selective agar (CNA); an additional 54.7% samples were recovered from Todd-Hewitt broth. Three genes (adhP, glnA and tkt) were sequenced successfully for six samples (1, 2. 4,6,12 and 65). The UPGMA tree with 1000 bootstrap showing the relationship between six samples was drawn.Conclusion: This study revealed that pregnant women of all ages are at risk of group B streptococcus colonization. Group B streptococcus was common among pregnant women at Dr George Mukhari Academic Hospital. No socio-economic risk factor was associated with group B streptococcus colonization. Results confirm that the combination of Todd-Hewitt broth and CNA agar plate is a time saving and sensitive method. The allelic profile, characteristics such as G+C (guanine+cytosine) content and dN/dS ratio were not analysed because of the smaller sample size used in this study, which shows that the MLST method was unsuccessful in this study. The UPGMA tree based on differences in consensus of the isolates showed that all group B streptococcus isolates are clustered and descend from a single node. / Life Sciences / M.Sc. (Life Sciences)