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Resumo: A leishmaniose visceral é uma enfermidade de grande impacto na saúde pública e de importância mundial que acomete o homem, animais silvestres e domésticos, como cães e gatos. A transmissão ocorre pela picada de flebotomídeos do gênero Lutzomya. Objetivou-se pesquisar a ocorrência de Leishmania (L.) chagasi em gatos provenientes de área endêmica para leishmaniose canina e humana pela utilização dos métodos diagnósticos: Reação de Imunofluorescência Indireta (RIFI), hemocultura e Reação em Cadeia da Polimerase (PCR) para Leishmania (L.) chagasi, comparar os resultados desses métodos quanto à especificidade e sensibilidade e realizar inquérito epidemiológico por meio de entrevista aos proprietários dos gatos avaliados. Foram colhidas amostras de sangue de 109 gatos provenientes de Birigui (SP), região endêmica para leishmaniose visceral canina e humana. Das 109 amostras de sangue, 52 (47,7%) foram positivas na hemocultura, três (2,7%) pela RIFI e 17 (15,6%) pela PCR. De acordo com os resultados encontrados, pode-se concluir que os gatos provenientes de áreas endêmicas para leishmaniose canina e humana são frequentemente expostos ao agente, podem se infectar, provavelmente desenvolver sintomatologia clínica e parasitemia. Faz-se também necessária a associação das técnicas sorológicas e moleculares para elevar a acuidade diagnóstica e, consequentemente, elucidar o verdadeiro papel do gato no ciclo epidemiológico da leishmaniose visceral / Abstract: The visceral leishmaniasis is a disease of great impact in the public health and of world significance. It affects man, wild and domestic animals, such as dogs and cats. The transmission occurs by the sting of phlebotomine vectors of the Lutzomyia gender. This work was aimed to study the occurrence of Leishmania (L.) chagasi in cats from an endemic area for canine and man leishmaniasis using Indirect Immunofluorescence Reaction (IFA), hemoculture and Polymerase Chain Reaction (PCR) for Leishmania (L.) chagasi diagnostic methods, compare the results of these methods as for the specificity and sensibility, and make an epidemiologic search through a questionnaire answered by the owners of the cats assessed. Blood samples were collected from 109 cats from Birigui (SP), an endemic region for canine and man visceral leishmaniasis. From 109 blood samples, 52 (47,7%) were positive in hemocultures, three (2,7%) in the IFA and 17 (15,6%) in the PCR. According to the results found one can conclude that cats from endemic areas for canine and man leishmaniasis are frequently exposed to de agent, they can infect themselves, develop a clinical simptomatology and parasithemia. It is also required an association of serological and molecular techniques to increase the diagnostic accuracy and, consequently, elucidate the true role of a cat in the epidemiological cycle of the visceral leishmaniasis / Orientador: Hélio Langoni / Coorientador: Simone Baldini Lucheis / Banca: Kátia Denise Saraiva Bresciani / Banca: Cassiano Victória / Mestre

Background and Objectives: Pertussis is an infectious disease caused by the Gram-negative bacterium Bordetella pertussis. In Peru, actual public health programs indicate that vaccination against B. pertussis must be mandatory and generalized, be-sides all detected cases must be reported. The objective of this study was to determine the prevalence of B. pertussis among children under five years of age with a presumptive diagnosis of whopping cough in Cajamarca, a region located in northern Peru. Materials and Methods: The population of this cross-sectional study were children under 5 years old hospitalized as presumptive cases of pertussis during December 2017 to December 2018. The nasopharyngeal samples were analyzed by real-time PCR for the detection of B. pertussis. Results: B. pertussis was identified as PCR + in 42.3% of our sample (33/78). The clinical presentation that was observed most frequently includes paroxysmal coughing (97%), difficulty breathing (69.7%), cyanosis (72.7%) and post-tussive em-esis (60.6%). Additionally, pneumonia was the most observed complication (33.3%). Four of the patients with PCR+ for B. pertussis presented only lymphocytosis, five only leukocytosis, two patients with decreased leukocytosis and lymphocytes and only one patient with leukopenia and relative lymphocytosis. There was a percentage of 84.8% of unvaccinated children in the PCR+ group. Finally, the mother was the most frequent symptom carrier (18.2%). Conclusion: In conclusion, in the studied population there is a high rate of PCR+ cases for B. pertussis. Laboratory values may show leukopenia or lymphopenia in patients with pertussis. It is necessary to use appropriate laboratory diagnostic tests in all infants with respiratory symptoms for B. pertussis. Since, the clinical diagnosis overestimates the diagnosis of pertussis. / Revisión por pares

The accuracy of the BioFire FilmArray Meningitis/Encephalitis (ME) panel for the identification of Cryptococcus has recently been called into question. The primary objective of this study was to assess the agreement between the BioFire ME polymerase chain reaction (PCR) and other markers of cryptococcal infection. This retrospective review identified five patients with cryptococcal meningoencephalitis, 4 of whom had a negative ME panel for Cryptococcus. All five cases had positive serum cryptococcal antigens, and three of five had a positive cerebrospinal fluid (CSF) culture for Cryptococcus. The BioFire ME panel does not appear to be reliable for ruling out Cryptococcus meningoencephalitis; multiple testing methods are recommended.

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Human papillomaviruses (HPV) have been implicated as etiologic agents in the genesis of cervical carcinoma and certain other benign lesions of the cervix. Clinical and epidemiological data, and the demonstration of HPV 16 viral DNA sequences in cervical cancer biopsies lend support to the etiologic association of HPV type 16 and cervical carcinoma. Interpretation of the association between HPV 16 and cervical cancer is limited by methods of detection. Different methods of detection of viral DNA sequences have been used based on DNA-DNA hybridization. Recently, a method based upon the in vitro enzymatic amplification of specific viral DNA sequences or polymerase chain reaction (PCR) has been used. The purpose of this study was to compare PCR with DNA-DNA hybridization methods in clinical specimens obtained from invasive cervical cancer. The in vitro enzymatic amplification or PCR was carried out on three specific regions of HPV 16. E6, E7 and L1 regions of HPV 16 were chosen as the target sequences of amplification and primers were synthesized specific to these regions. PCR was performed on 163 cervical cancer specimens using primers specific for E6 and E7 regions of HPV 16. 112 of these specimens were also analyzed using L1 primers of HPV 16. Estimates of sensitivity and specificity of the different methods to see if PCR is a better, more sensitive method compared to the other methods were computed. The results suggest that although percent positivity by PCR method increases significantly, thereby improving sensitivity of detection, the specificity suffers compared to the other methods. However the advantages of using PCR as a diagnostic tool are attractive, as it requires only picogram quantities of DNA, is rapid and easy to perform, and is amenable to automation. / Thesis / Master of Science (MS)

Latent feline leukemia virus (FeLV) infections, in which proviral DNA is integrated into host DNA, but not actively transcribed, are suspected to be associated with many diseases. Bone marrow is the suspected site of the majority of latent infections. The purpose of this study was to determine if polymerase chain reaction (PCR) could detect FeLV proviral DNA in bone marrow and provide a method of detecting latent infections. Blood and bone marrow samples from fifty cats and bone marrow from one fetus were collected; sixteen had FeLV-associated diseases. Serum ELISA, blood and bone marrow immunofluorescent antibody test (IFA), and blood and bone marrow PCR were performed on each cat, and IFA and PCR on bone marrow of the fetus. Forty-one cats were FeLV negative. Five cats and one fetus were persistently infected with FeLV. Four cats were discordant; two ELISA positive with other tests negative, one bone marrow IFA negative with other tests positive, and one bone marrow IFA positive with other tests negative. No cats were positive on bone marrow PCR only. These results indicate that PCR can detect FeLV in bone marrow, but no cats in this study harbored FeLV only in the bone marrow. Not all cats with FeLV-associated diseases are persistently or latently infected with FeLV. / Master of Science

Corneal transplantation carries a high rate of success in the domestic cat and is an indicated treatment for specific corneal diseases in this species. The potential for iatrogenic transmission of viral diseases is a well-recognized problem in human corneal transplantation programs and screening donors for certain diseases is routine. Feline leukemia virus (FeLV) is a common agent of disease in domestic cats and available blood tests are highly effective in identification of infected individuals. This study investigates the presence of FeLV within corneal tissues of FeLV infected cats. Seventeen cats were identified to be positive for serum p27 antigen by enzyme-linked immunosorbent assay (ELISA). Twelve of these individuals were found to be positive on peripheral blood by immunofluorescent antibody (IFA) testing. Seventeen ELISA negative cats were identified to serve as negative controls. Full thickness corneal specimens were collected from all subjects and analyzed for the presence of FeLV proviral DNA and gp70 antigen by polymerase chain reaction (PCR) and immunohistochemical (IHC) testing, respectively. Eleven (64.7%) positive corneal PCR results were obtained from 17 ELISA positive cats. Of 12 cats which were both ELISA and IFA positive on peripheral blood, 10 (83.3%) had positive corneal PCR results. All corneal tissues from ELISA negative subjects were PCR negative. IHC staining of corneal sections revealed the presence of FeLV gp70 in corneal tissues of nine (52.9%) ELISA positive cats. Of the 12 cats which were both ELISA and IFA positive on peripheral blood, 8 (66.7%) had positive corneal IHC results. Positive IHC staining was localized to the corneal epithelium. Corneal tissues of all ELISA negative cats and all IFA negative cats were negative on IHC testing. This study reveals FeLV to be present within the corneal epithelium of some FeLV infected cats. Screening potential corneal donors for this virus is warranted. This work was funded by grants from the American College of Veterinary Ophthalmologists, the Virginia Veterinary Medical Association Pet Memorial Fund, and the DSACS Quick Response Fund. / Master of Science