Detection of pathogens from periodontal lesions

OBJECTIVE: To comparatively detect A. actinomycetemcomitans and F. nucleatum from periodontal and healthy sites. METHODS: Subgingival clinical samples from 50 periodontitis adult patients and 50 healthy subjects were analyzed. Both organisms were isolated using a trypticase soy agar-bacitracin-vancomycin (TSBV) medium and detected by PCR. Conventional biochemical tests were used for bacteria identification. RESULTS: A. actinomycetemcomitans and F. nucleatum were isolated in 18% and 20% of the patients, respectively, and in 2% and 24% of healthy subjects. Among A. actinomycetemcomitans isolates, biotype II was the most prevalent. Primer pair AA was 100% sensitive in the detection of A. actinomycetemcomitans from both subject groups. Primers ASH and FU were also 100% sensitive to detect this organism in healthy subject samples. Primer pair FN5047 was more sensitive to detect F. nucleatum in patients or in healthy samples than primer 5059S. Primers ASH and 5059S were more specific in the detection of A. actinomycetemcomitans and F. nucleatum, respectively, in patients and in healthy subject samples. CONCLUSIONS: PCR is an effective tool for detecting periodontal pathogens in subgingival samples, providing a faster and safer diagnostic tool of periodontal diseases. The method's sensitivity and specificity is conditioned by the choice of the set of primers used.

Prevalence of putative periodontopathogens from periodontal patients and healthy subjects in São Paulo, SP, Brazil

A. actinomycetemcomitans, B. forsythus, P. gingivalis, C. rectus, E. corrodens, P. intermedia, F. nucleatum, and T. denticola were identified from subgingival plaque from 50 periodontal patients and 50 healthy subjects. PCR products from each species showed a specific band and could be used to identify periodontal organisms from clinical specimens. Identical negative or positive results between PCR and culture occurred in 66% (A. actinomycetemcomitans) to 93% (F. nucleatum) of the samples. PCR detection odds ratio values for A. actinomycetemcomitans, B. forsythus, C. rectus, E. corrodens, P. intermedia, and T. denticola were significantly associated with disease having a higher OR values for B. forsythus (2.97, 95% CI 1.88 - 4.70). Cultures showed that A. actinomycetemcomitans, B. forsythus and P. intermedia were associated with periodontitis, however, P. gingivalis, C. rectus, E. corrodens and F. nucleatum were not significantly associated with the disease.

Antimicrobial peptides and nitric oxide production by neutrophils from periodontitis subjects

Neutrophils play an important role in periodontitis by producing nitric oxide (NO) and antimicrobial peptides, molecules with microbicidal activity via oxygen-dependent and -independent mechanisms, respectively. It is unknown whether variation in the production of antimicrobial peptides such as LL-37, human neutrophil peptides (HNP) 1-3, and NO by neutrophils influences the pathogenesis of periodontal diseases. We compared the production of these peptides and NO by lipopolysaccharide (LPS)-stimulated neutrophils isolated from healthy subjects and from patients with periodontitis. Peripheral blood neutrophils were cultured with or without Aggregatibacter actinomycetemcomitans-LPS (Aa-LPS), Porphyromonas gingivalis-LPS (Pg-LPS) and Escherichia coli-LPS (Ec-LPS). qRT-PCR was used to determine quantities of HNP 1-3 and LL-37 mRNA in neutrophils. Amounts of HNP 1-3 and LL-37 proteins in the cell culture supernatants were also determined by ELISA. In addition, NO levels in neutrophil culture supernatants were quantitated by the Griess reaction. Neutrophils from periodontitis patients cultured with Aa-LPS, Pg-LPS and Ec-LPS expressed higher HNP 1-3 mRNA than neutrophils from healthy subjects. LL-37 mRNA expression was higher in neutrophils from patients stimulated with Aa-LPS. Neutrophils from periodontitis patients produced significantly higher LL-37 protein levels than neutrophils from healthy subjects when stimulated with Pg-LPS and Ec-LPS, but no difference was observed in HNP 1-3 production. Neutrophils from periodontitis patients cultured or not with Pg-LPS and Ec-LPS produced significantly lower NO levels than neutrophils from healthy subjects. The significant differences in the production of LL-37 and NO between neutrophils from healthy and periodontitis subjects indicate that production of these molecules might influence individual susceptibility to important periodontal pathogens.

Antimicrobial activity of the essential oil from Lippia sidoides, carvacrol and thymol against oral pathogens

Dental caries and periodontal disease are associated with oral pathogens. Several plant derivatives have been evaluated with respect to their antimicrobial effects against such pathogenic microorganisms. Lippia sidoides Cham (Verbenaceae), popularly known as "Alecrim-pimenta" is a typical shrub commonly found in the Northeast of Brazil. Many plant species belonging to the genus Lippia yield very fragrant essential oils of potential economic value which are used by the industry for the commercial production of perfumes, creams, lotions, and deodorants. Since the leaves of L. sidoides are also extensively used in popular medicine for the treatment of skin wounds and cuts, the objective of the present study was to evaluate the composition and antimicrobial activity of L. sidoides essential oil. The essential oil was obtained by hydro-distillation and analyzed by GC-MS. Twelve compounds were characterized, having as major constituents thymol (56.7%) and carvacrol (16.7%). The antimicrobial activity of the oil and the major components was tested against cariogenic bacterial species of the genus Streptococcus as well as Candida albicans using the broth dilution and disk diffusion assays. The essential oil and its major components thymol and carvacrol exhibited potent antimicrobial activity against the organisms tested with minimum inhibitory concentrations ranging from 0.625 to 10.0 mg/mL. The most sensitive microorganisms were C. albicans and Streptococcus mutans. The essential oil of L. sidoides and its major components exert promising antimicrobial effects against oral pathogens and suggest its likely usefulness to combat oral microbial growth.

Prevalence of periodontal disease in dogs and owners' level of awareness - a prospective clinical trial

Periodontal disease (PD) is widely known among veterinarians for its high prevalence and serious consequences to the dogs. The objective of this study was to assess the occurrence of PD in dogs that live in the micro-region of Viçosa, treated at the Veterinary Hospital of the Federal University of Viçosa (HVT - Hospital Veterinário da Universidade Federal de Viçosa), as well as to assess how aware of this disease dog owners are. In order to do so, all dogs treated at the HVT from March 10th, 2009 to November 30th, 2009, on alternate days, had their oral cavities examined. Medical history data, such as age, type of food, main complaint and owner consent, halitosis, presence of dental calculus, inflammation and gingival recession and tooth loss, were collected. A prevalence of 88.67% was found for PD in dogs referred to the HVT, and 2.67% were referred due to this disease. Of all the owners who participated in the study, 43.83% knew about periodontal disease and of these 17.46% made use of some type of prevention or treatment. Therefore, periodontal disease is highly prevalent and the owners are not aware of the disease. Thus, a dog owner clarification program on periodontal disease is needed in the area where HVT-UFV operates.

Registro periodontal simplificado em gestantes

OBJETIVO: Avaliar através do PSR (Registro Periodontal Simplificado) a prevalência, severidade e necessidades básicas de tratamento da doença periodontal em gestantes que freqüentaram a Clínica de Prevenção da Faculdade de Odontologia de Araraquara ­ UNESP. MÉTODOS: Foram examinadas 41 gestantes com idades que variaram de 16 a 37 anos. O PSR foi aplicado com auxílio de uma sonda especialmente recomendada para este exame (sonda Trinity - tipo 621 OMS), indicando os códigos 0 a 4 cujos critérios identificam de saúde gengival, sangramento, cálculo, bolsa periodontal rasa e profunda. Estes foram atribuídos a cada sextante, podendo ou não estarem associados a um asterisco (*) diante da presença de recessão gengival, invasão de furca, mobilidade ou alterações muco-gengivais. RESULTADOS: Demonstraram que 100% das gestantes apresentaram alguma alteração gengival, sendo os códigos 2 (56,1%) e o * (19,5%) os mais prevalentes. Os grupos etários de 15-19 e 20-24 anos, apresentaram o código 2 como maior escore e ausência de sextante excluído (X). A partir do grupo de 25-29 anos, além da maior prevalência ainda ser do código 2 (54,5%), ocorreram os códigos 3 e 4 (bolsa periodontal). Os códigos * e sextante excluído (X) tenderam a aumentar com a idade no grupo de 30-37 anos. De modo geral, os códigos 1 e 2, prevaleceram em relação ao percentual de sextantes afetados, correspondendo a 41,6% e 39,8%, respectivamente e afetando 2,49 e 2,39 sextantes, em média, por gestante. Em relação às necessidades de tratamento, 90,2% das gestantes necessitaram tratamentos adicionais aos preventivos, ou seja, 61,0% das gestantes necessitaram de raspagem e alisamento radicular e/ou eliminar margens de restaurações defeituosas e 29,2% de tratamento complexo. CONCLUSÃO: O atendimento às necessidades de tratamento na gravidez deve receber especial atenção com o intuito de se promover saúde bucal e motivação, e conseqüentemente, contribuir para minimizar a provável transmissibilidade de microrganismos bucais patogênicos para a criança, obtendo assim uma prevenção primária das principais doenças bucais.

Doença periodontal materna como fator associado ao baixo peso ao nascer

OBJETIVO: Estudos recentes têm apresentado evidências de que a doença periodontal em gestantes pode ser um dos determinantes do baixo peso ao nascer. Realizou-se estudo para verificar a existência de associação entre doença periodontal materna e baixo peso ao nascer. MÉTODOS: Estudo do tipo caso-controle com 302 mulheres, sendo 102 mães de nascidos vivos de baixo peso (grupo caso) e 200 mães de nascidos vivos com peso normal (grupo controle). A existência de associação entre doença periodontal e baixo peso ao nascer foi avaliada mediante modelo multivariado de regressão logística, considerando outros fatores de risco para o baixo peso. RESULTADOS: Ambos os grupos de mães eram comparáveis no que se refere a idade, altura, peso pré-gestacional, tabagismo, alcoolismo, doenças prévias, estado civil, situação socioeconômica, número de escovações e uso de fio dental, número de refeições diárias, e visitas ao dentista. A doença periodontal foi diagnosticada em 57,8% das mães do grupo caso e 39,0% do grupo controle. A análise de regressão logística indicou associação positiva entre doença periodontal e baixo peso ao nascer (ORbruto=2,15; IC 95%: 1,32-3,48), especialmente entre as mães com escolaridade menor ou igual a quatro anos (ORajustada=3,98; IC 95%: 1,58-10,10). CONCLUSÕES: A doença periodontal é um possível fator de risco para o baixo peso ao nascer.

Desigualdades sociais e doença periodontal no estudo SBBrasil 2010: abordagem multinível

OBJETIVO: Estimar a prevalência e a distribuição geográfica da doença periodontal na população adulta brasileira e sua associação com desigualdades sociais contextuais e individuais. MÉTODOS: Foram utilizados dados dos adultos de 35 a 44 anos de idade da Pesquisa Nacional de Saúde Bucal – SBBrasil 2010 (n = 9.564). O Índice Periodontal Comunitário (CPI) e o Índice de Perda Periodontal (PIP) foram usados para definir a doença periodontal em “moderada a grave” (CPI > 2 e PIP > 0) e “grave” (CPI > 2 e PIP > 1). As desigualdades sociais contextuais incluíram o índice de desenvolvimento humano e a desigualdade de renda (Índice de Gini). Outras variáveis contextuais foram a cobertura de equipes de saúde bucal da Estratégia de Saúde da Família e o percentual de adultos fumantes. Modelos de regressão logística multinível para os participantes com dados completos (n = 4.594) foram usados para estimar odds ratios (OR) e intervalos de 95% de confiança (IC95%) entre desigualdades sociais e doença periodontal. RESULTADOS: A prevalência da doença periodontal “moderada a grave” em brasileiros adultos foi de 15,3% e 5,8% para a condição “grave”, com variações consideráveis entre os municípios. Dentre as variáveis contextuais, a desigualdade de renda foi independentemente associada com a doença periodontal “grave” (OR = 3,0; IC95% 1,5;5,9). A menor cobertura de equipes de saúde bucal foi associada com as duas formas de doença periodontal, enquanto o percentual de fumantes manteve-se associado com a doença periodontal “moderada a grave”. Adultos com idade mais avançada, de cor de pele parda, sexo masculino, menor renda familiar e menor escolaridade apresentaram maiores chances para ambas as condições periodontais investigadas. CONCLUSÕES: No Brasil, a prevalência da doença periodontal variou conforme o município e a definição de doença empregada. A desigualdade de renda teve um papel importante na ocorrência da doença periodontal “grave”. Características individuais de posição social foram associadas com as duas formas de doença periodontal.

Seroprevalence of HIV, HTLV-I/II and other perinatally-transmitted pathogens in Salvador, Bahia

Generation of epidemiological data on perinatally-transmitted infections is a fundamental tool for the formulation of health policies. In Brazil, this information is scarce, particularly in Northeast, the poorest region of the country. In order to gain some insights of the problem we studied the seroprevalence of some perinatally-transmitted infections in 1,024 low income pregnant women in Salvador, Bahia. The prevalences were as follow: HIV-1 (0.10%), HTLV-I/II (0.88%), T.cruzi (2.34%). T.pallidum (3.91%), rubella virus (77.44%). T.gondii IgM (2.87%) and IgG (69.34%), HBs Ag (0.6%) and anti-HBs (7.62%). Rubella virus and T.gondii IgG antibodies were present in more than two thirds of pregnant women but antibodies against other pathogens were present at much lower rates. We found that the prevalence of HTLV-I/II was nine times higher than that found for HIV-1. In some cases such as T.cruzi and hepatitis B infection there was a decrease in the prevalence over the years. On the other hand, there was an increase in the seroprevalence of T.gondii infection. Our data strongly recommend mandatory screening tests for HTLV-I/II, T.gondii (IgM), T.pallidum and rubella virus in prenatal routine for pregnant women in Salvador. Screening test for T.cruzi, hepatitis and HIV-1 is recommended whenever risk factors associated with these infections are suspected. However in areas with high prevalence for these infections, the mandatory screening test in prenatal care should be considered.

Periodontal aspects of the juvenile form of paracoccidioidomycosis

Three cases of the juvenile form of paracoccidioidomycosis are reported. Emphasis has been given to the oral manifestations, particularly the periodontal involvement. The main periodontal findings were: generalized and progressive alveolar bone destruction leading to gingival recession with exposure of the tooth roots, and spontaneous tooth losses. The gingival mucosa was predominantly smooth, erithematous and slightly swollen. These aspects, although rare, may be the earliest signs of the disease and sometimes its only manifestation.

FREQUENCY, URINALYSIS AND SUSCEPTIBILITY PROFILE OF PATHOGENS CAUSING URINARY TRACT INFECTIONS IN ENUGU STATE, SOUTHEAST NIGERIA

Objective: This study was designed to determine the frequency and causative agent(s) of urinary tract infections (UTIs) in individuals with symptoms of urinary tract infections in Enugu State of Southeast Nigeria, and to determine the antibiotic susceptibility pattern of microbial agents isolated from urine culture.Methods: The study involved 211 individuals (149 females and 62 males) clinically suspected for UTI. Urine samples were collected by the mid-stream ‘clean catch’ method and tested using standard procedures. Antibiotic susceptibility of the isolated pathogens was tested using the Kirby-Bauer technique according to the Clinical and Laboratory Standards Institute (CLSI) guidelines.Results: Microscopy of centrifuged urine samples showed 16 patients had pyuria while 54 had pus cells. Calcium oxalate crystals were found in 14 samples. Urinalysis performed with urine samples showed 17 had protein; seven were nitrite positive and three had moderate to high glucose concentration. Fifty-four urine samples (36.2%) from females and 12 (19.4%) from males showed significant growth upon culture. Gram stain and biochemical tests identified nine different organisms with Escherichia coli as the most common isolated species. Forty three randomly selected strains were further tested for their susceptibility against a panel of antibiotics. Thirty isolates (81.08%) were resistant to four or more antibiotics with the highest resistance shown by E. coli (76.67%). All the Gram- negative isolates were resistant to Ampicilox, Cefuroxime and Amoxicillin.Conclusion: Urinary tract infections were found more in females in the area under study. As found in other studies, E. coli was the most predominant isolate, although other organisms seem to be on the increase.

Report on ticks collected in the Southeast and Mid-West regions of Brazil: analyzing the potential transmission of tick-borne pathogens to man

Specimens of ticks were collected in 1993, 1996, 1997, and 1998, mostly from wild and domestic animals in the Southeast and Mid-West regions of Brazil. Nine species of Amblyommidae were identified: Anocentor nitens, Amblyomma cajennense, Amblyomma ovale, Amblyomma fulvum, Amblyomma striatum, Amblyomma rotundatum, Boophilus microplus, Boophilus annulatus, and Rhipicephalus sanguineus. The potential of these tick species as transmitters of pathogens to man was analyzed. A Flaviviridade Flavivirus was isolated from Amblyomma cajennense specimens collected from a sick capybara (Hydrochaeris hydrochaeris). Amblyomma cajennense is the main transmitter of Rickettsia rickettsii (=R. rickettsi), the causative agent of spotted fever in Brazil. Wild mammals, mainly capybaras and deer, infested by ticks and living in close contact with cattle, horses and dogs, offer the risk of transmission of wild zoonosis to these domestic animals and to man.

Mycetoma fungal infection: multiple organisms as colonizers or pathogens?

We describe a patient with mycetoma or Madura foot, in which histopathological stains of the bone and surface cultures suggested three different organisms including Nocardia species as the cause. Criteria for the diagnosis of the organisms, differentiation between colonizer and pathogen, and significance of mixed infections are discussed.