Usefulness of catheter tip culture in the diagnosis of neonatal infections

Objective: To determine the number of colony-forming units (CFU) that best correlates with catheter-related infections (CRI) in newborns.Methods: This was a prospective study of semiquantitative cultures of catheter tips obtained from newborns in the neonatal unit at Faculdade de Medicina de Botucatu, state of São Paulo, Brazil. The microorganisms isolated from catheter and peripheral blood cultures were identified and submitted to a drug susceptibility test. The optimal cutoff point was determined by the receiver operating characteristic (ROC) curve.Results: A total of 85 catheters obtained from 63 newborns were studied. Staphylococcus epidermidis was the predominant species in the catheters (75%). Eight of 11 (72.7%) CRI episodes were associated with coagulase-negative staphylococci, six of which were of the S. epidermidis type. ROC curve analysis indicated that the optimal cutoff point for the diagnosis of CRI was 122 CFU.Conclusions: The cutoff point of 122 CFU correlated best with the diagnosis of CRI in newborns.

What is your diagnosis? Hepatosplenomegaly in a howler monkey (Alouatta fusca)

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Comparison between procalcitonin and C-reactive protein for early diagnosis of children with sepsis or septic shock

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Diagnosis of clinical bovine mastitis by fine needle aspiration followed by staining and scanning electron microscopy in a Prototheca zopfii outbreak

Biopsy by fine needle aspiration together with microbiological examination and scanning electron microscopy were evaluated in diagnosis of clinical bovine mastitis in a Prototheca zopfii outbreak. Fine needle aspiration was performed in 21 mammary quarters from ten Holstein cows presenting clinical mastitis caused by P. zopfii. The algae were previously identified in the microbiological examination of milk collected from these cows. Material aspirated from these 21 mammary glands was submitted to cytological staining (Gram, Giemsa and/or Shor staining). Fine needle aspiration enabled cytological identification of the algae in these 21 mammary glands, from which P. zopfii was isolated in the milk. Simultaneously, five mammary fragments collected by fine needle aspiration from these 21 mammary glands presenting clinical mastitis were also submitted to microbiological examination. P. zopfii was also isolated from these five fragments. Scanning electron microscopy technique also identified three of these five P. zopfii strains isolated from mammary fragments collected by cytological aspiration. These results suggest that fine needle aspiration may be an alternative method for the diagnosis of clinical mastitis.

Detection of IgG in cerebrospinal fluid for diagnosis of neurocysticercosis: evaluation of saline and SDS extracts from Taenia solium and Taenia crassiceps metacestodes by ELISA and immunoblot assay

We compared saline (S) and sodium dodecyl sulphate (SDS) extracts from Taenia solium (homologous species - HO) and Taenia crassiceps (heterologous species - HE) metacestodes in order to detect Ige by ELISA and immunoblot assay (IBA) in cerebrospinal fluid (CSF) for the diagnosis of human neurocysticercosis (NC). CSF samples were obtained from 93 patients. of these, 40 had NC, five had a diagnosis of probable NC, nine had central nervous system schistosomiasis or strongyloidiasis and 39 had other neurological alterations. Samples were analysed by ELISA and the results were compared with IBA in all samples with confirmed and probable NC diagnosis, in all samples with other central nervous system parasitic infection, and in 10 of those with another neurological alterations. ELISA sensitivity was 100%, 85%, 95% and 87.5% for the S-HO, S-HE, SDS-HO and SDS-HE extracts, respectively, and ELISA specificity was 100% for S-HO, S-HE, SDS-HO extracts and 97.9% for SDS-HE antigen. Immunodominant peptides detected by IBA were, by decreasing percentage of recognition: 64-68 and 45 kDa for S-HO; 108-114, 92-95, 64-68, 83 and 88 kDa for S-HE; 64-68, 108-114, 77 and 86 kDa for SDS-HO; and 108-114, 88 and 92-95 kDa for SDS-HE. Overall the homologous antigenic extracts showed higher sensitivity than the heterologous extracts in the diagnosis of NC in CSF samples. The heterologous extracts contained most of the immunodominant peptides presented in the homologous extracts, which are recognized by Ige antibodies in CSF samples.

Epidemiological and Serological Aspects in Canine Toxoplasmosis in Animals with Nervous Symptoms

The presence of anti-Toxoplasma gondii IgM and IgG antibodies was studied in samples of blood serum taken from eighty dogs with nervous symptoms at the Serviço de Enfermidades Infecciosas dos Animais, Faculdade de Medicina Veterinária e Zootecnia, Unesp, Botucatu, São Paulo, Brazil. The frequency of IgG titers were 16 (13.7%), 64 (13.7%), and 256 (5%), and for IgM titers were 16 (7.5%), 64 (15%), and 256 (8.7%). Positive reactions were more frequent in the older animals, males, from a rural environment, in constant contact with small animals, principally birds and rodents. There was a higher frequency of a positive reaction in dogs fed with kitchen food, especially in those fed with raw ingredients. The most common neurological pictures were alterations in consciousness, in movement, and in the hand-cart test. The percentage of reagents with specific IgM antibodies was high, indicating active infections, but the possibility of co-infection with the distemper virus can not be discarded, and this may be a predisposing factor for toxoplasmosis infection, once the distemper virus has a potent immunosupressive action.

Sorologia Anti PGL-1 e risco de ocorrência de hanseníase em área de alta endemicidade do Estado de São Paulo: quatro anos de seguimento

Os testes sorológicos para diagnóstico de hanseníase, usando o glicolipídeo-fenólico-1 (PGL-1), considerado antígeno específico do M. leprae, têm aberto algumas possibilidades de estudo do comportamento epidemiológico desta doença. Algumas questões, como tempo de latência da doença, infecção subclínica e importância do contato intra-domiciliar (contatos) no controle da endemia, puderam ser melhor analisadas usando este instrumental. Este estudo teve por objetivo verificar a existência de associação entre a situação sorológica e a ocorrência de hanseníase. Foram seguidas, durante 4 anos, 6.520 pessoas com idade igual ou superior a 5 anos, submetidas no início do seguimento ao teste sorológico Anti PGL-1, pertencentes ao universo de 7.416 habitantes da área urbana de um município paulista caracterizado por elevada endemicidade de hanseníase. Foi identificado um grupo de 590 indivíduos soropositivos (9,0 %). Foram diagnosticados, no período, 82 casos novos de hanseníase, 26 no grupo de soropositivos (441 casos novos/10.000 indivíduos) e 48 no de soronegativos (81/10.000). Entre os que não fizeram sorologia, surgiram 8 casos novos (89/10.000). Procurou-se controlar, na análise, a condição de contato, dado que a taxa de soropositividade padronizada por idade e sexo era de 9,61% no grupo de contatos e 7,65% no de não-contatos. Tomando-se os não-contatos soronegativos como o grupo de não expostos, foram calculados os riscos relativos de adoecimento no período, a partir das taxas de detecção padronizadas por idade, resultando no seguinte: os contatos ID soropositivos apresentaram a taxa de 1.704/10.000, 27 vezes maior que a dos não-expostos, igual a 63/10.000; os não-contatos soropositivos e os contatos soronegativos apresentaram taxas, respectivamente, de 274 e 198/10.000, ambas maiores que as dos não-expostos e iguais entre si. A soropositividade associou-se à elevação de 8,6 vezes do risco de hanseníase entre os contatos e de 4,4 entre os não-contatos. Na situação epidemiológica estudada, caracterizada por elevada endemicidade de hanseníase, 50% dos casos novos surgiram entre os não-contatos soronegativos, ou seja, sem fonte de infecção conhecida. Portanto, o teste anti-PGL-1 usado revela-se, na prática, de pouca aplicabilidade. Resta estudar ainda o comportamento da sorologia anti-PGL-1 em áreas de média e baixa endemicidade para que se possa tirar conclusões mais consubstanciadas sobre sua utilidade no controle da endemia. Recomenda-se o aprofundamento das pesquisas sorológicas e de outras que aprimorem o diagnóstico precoce da infecção subclínica, inclusive para detecção de formas paucibacilares, para se ampliar as possibilidades de influir no controle endêmico.

A polymerase chain reaction for the detection of Brucella canis in semen of naturally infected dogs

The objective was to evaluate a PCR assay for the detection of Brucella canis in canine semen, comparing its performance with that of bacterial isolation, serological tests and PCR assay of blood. Fifty-two male dogs were examined clinically to detect reproductive abnormalities and their serum was tested by the rapid slide agglutination test, with and without 2-mercaptoethanol (2ME-RSAT and RSAT, respectively). In addition, microbiological culture and PCR assays were performed on blood and semen samples. The findings of the semen PCR were compared (Kappa coefficient and McNemar test) to those of blood PCR, culture of blood and semen, RSAT, and 2ME-RSAT. Nucleic acid extracts from semen collected from dogs not infected with B. canis were spiked with decreasing amounts of B. canis RM6/66 DNA and the resulting samples subjected to PCR. In addition, semen samples of non-infected dogs were spiked with decreasing amounts of B. canis CFU and the resulting suspensions were used for DNA extraction and amplification. of the 52 dogs that were examined, the following tests were positive: RSAT, 16 (30.7%); 2ME-RSAT, 5 (9.6%); blood culture, 14 (26.9%); semen culture, 11 (21.1%); blood PCR, 18 (34.6%); semen PCR, 18 (34.6%). The PCR assay detected as few as 3.8 fg of B. canis DNA experimentally diluted in 444.9 ng of canine DNA (extracted from semen samples of noninfected dogs). In addition, the PCR assay amplified B. canis genetic sequences from semen samples containing as little as 1.0 x 10(0) cfu/mL. We concluded that PCR assay of semen was a good candidate as a confirmatory test for the diagnosis of brucellosis in dogs; its diagnostic performance was similar to blood culture or blood PCR. Furthermore, the PCR assay of semen was more sensitive than the 2ME-RSAT or semen culture. Examination of semen by PCR should be included for diagnosis of brucellosis prior to natural mating or AI; in that regard, some dogs that were negative on serological and microbiological examinations as well as blood PCR were positive on PCR of semen. (c) 2007 Elsevier B.V. All rights reserved.

A polymerase chain reaction for detection of Brucella canis in vaginal swabs of naturally infected bitches

A PCR assay for the detection of Brucella canis in canine vaginal swab samples was evaluated, comparing its performance with that of bacterial isolation, serological tests, and a blood PCR assay. One hundred and forty-four female dogs were clinically examined to detect reproductive problems and they were tested by the rapid slide agglutination test, with and without 2-mercaptoethanol (2ME-RSAT and RSAT, respectively). In addition, microbiological culture and PCR were performed on blood and vaginal swab samples. The results of the vaginal swab PCR were compared to those of the other tests using the Kappa coefficient and McNemar test. of the 144 females that were examined, 66 (45.8%) were RSAT positive, 23 (15.9%) were 2ME-RSAT positive, 49 (34.02%) were blood culture positive, 6 (4.1%) were vaginal swab culture positive, 54 (37.5%) were blood PCR positive, 52 (36.2%) were vaginal swab PCR positive, and 50.69% (73/144) were positive by the combined PCR. The PCR was able to detect as few as 3.8 fg of B. canis DNA experimentally diluted in 54 ng of canine DNA, extracted from vaginal swab samples of non-infected bitches. In addition, the PCR assay amplified B. canis genetic sequences from vaginal swab samples containing 1.0 x 10(0) cfu/mL. In conclusion, vaginal swab PCR was a good candidate as a confirmatory test for brucellosis diagnosis in bitches suspected to be infected, especially those negative on blood culture or blood PCR; these animals may be important reservoirs of infection and could complicate attempts to eradicate the disease in confined populations. (C) 2007 Elsevier B.V. All rights reserved.