RESUMO
Human cryptosporidiosis is an intestinal infection caused by different species belonging to the genus Cryptosporidium in both immunocompetent and immunocompromised individuals. The life cycle of Cryptosporidium sp. when affecting the digestive system is well known but the infection of other organs is less studied. Molecular methods are necessary for species and subtypes identification. The goal of this work is to propose a new approach that contributes to the diagnosis of the extra-intestinal dissemination process of Cryptosporidium infection. Cryptosporidium sp. was detected in stool and biopsy samples of two HIV-infected patients. DNA was extracted from feces, biopsy specimens, blood, and cerebrospinal fluid (CSF). All samples were analyzed by nested PCR-RFLP of the 18S rDNA, real-time PCR, and gp60 subtyping. Cryptosporidium DNA was detected in stool and tissue samples and it was also present in blood and CSF samples. Both cases were characterized as Cryptosporidium hominis subtype IeA11G3T3. This is the first report that demonstrates the presence of Cryptosporidium DNA in blood and CSF of HIV-infected patients.
Assuntos
Criptosporidiose/diagnóstico , Cryptosporidium/isolamento & purificação , DNA de Protozoário/sangue , DNA de Protozoário/líquido cefalorraquidiano , Infecções por HIV/complicações , Adulto , Animais , Criptosporidiose/sangue , Criptosporidiose/líquido cefalorraquidiano , Criptosporidiose/complicações , Cryptosporidium/classificação , Cryptosporidium/genética , DNA Ribossômico/genética , Fezes/química , Infecções por HIV/sangue , Infecções por HIV/líquido cefalorraquidiano , Infecções por HIV/parasitologia , Humanos , Hospedeiro Imunocomprometido , Masculino , Pessoa de Meia-Idade , Polimorfismo de Fragmento de Restrição , Reação em Cadeia da Polimerase em Tempo RealRESUMO
Infection by Cryptosporidium serpentis is one of the most important diseases in reptiles and is characterized by chronic clinical or subclinical infection and the presence of hypertrophic gastritis, food regurgitation, progressive weight loss, mortality, and intermittent or continuous shedding of oocysts in the feces. The objectives of this study were to standardize an indirect enzyme-linked immunosorbent assay (ELISA) to detect antibodies against C. serpentis and to evaluate the clinical, parasitological, and humoral immune response in snakes naturally infected with C. serpentis. Twenty-one snakes naturally infected with C. serpentis and housed at the Butantan Institute, São Paulo, Brazil, underwent clinical and parasitological analyses for C. serpentis infection through daily records of clinical signs and a monthly survey of fecal shedding of oocysts using the Kinyoun's acid-fast staining. The serological evaluation was performed monthly by indirect ELISA using crude total antigen from oocysts of C. serpentis to detect anti-C. serpentis antibodies. Clinical symptoms consisted of food regurgitation, inappetence, and progressive weight loss. The parasitological analysis revealed intermittent fecal shedding of a variable number of oocysts in all snakes, with positivity in 85.32% (157/184) of the samples. The indirect ELISA was positive in 68.25% (86/126) of the samples. A humoral immune response was observed in most animals; however, fluctuating antibodies levels, leading to alternating positive and negative results, were observed in most snakes.
Assuntos
Criptosporidiose/veterinária , Cryptosporidium/classificação , Serpentes , Animais , Criptosporidiose/sangue , Criptosporidiose/parasitologia , Serpentes/classificaçãoRESUMO
BACKGROUND: Mannose-binding lectin (MBL) is a component of the innate immune response and binds microbial surfaces through carbohydrate recognition domains. MBL deficiency may contribute to susceptibility to a variety of infectious diseases, particularly in young children. MBL binds to the Cryptosporidium sporozoite and may be important in resistance to cryptosporidiosis. METHODS: We studied the association of serum MBL levels and cryptosporidiosis in a case-control study of young Haitian children with cryptosporidiosis versus children who were control subjects. RESULTS: Ninety-nine children were enrolled, as follows: 49 children with cryptosporidiosis, 41 healthy controls, and 9 children with diarrhea from other causes. Case children were more malnourished than controls, and 49% had persistent or chronic diarrhea. At enrollment, mean serum MBL levels were markedly lower in children with cryptosporidiosis (P = .002), as was the number of children with an MBL deficiency of < or = 70 ng/mL (P = .005). In multivariate analysis, the association of cryptosporidiosis and MBL deficiency persisted (P = .002; adjusted odds ratio, 22.4), as did the association of cryptosporidiosis with general malnutrition. The subset of children with cryptosporidiosis and MBL deficiency were more likely to be male (P = .025). CONCLUSIONS: MBL may be an important component of innate immune protection against Cryptosporidium infection in young children. Additional studies are necessary to determine whether MBL intestinal losses, deficient epithelial expression, and/or genetic polymorphisms in the MBL gene contribute to MBL deficiency in cryptosporidiosis and other enteric infections in young children.
Assuntos
Criptosporidiose/metabolismo , Lectina de Ligação a Manose/deficiência , Estudos de Casos e Controles , Criptosporidiose/sangue , Criptosporidiose/imunologia , Suscetibilidade a Doenças , Feminino , Haiti , Humanos , Imunidade Inata/fisiologia , Lactente , Masculino , Lectina de Ligação a Manose/sangue , Lectina de Ligação a Manose/imunologiaAssuntos
Criptosporidiose/metabolismo , Lectina de Ligação a Manose/deficiência , Estudos de Casos e Controles , Criptosporidiose/sangue , Criptosporidiose/genética , Criptosporidiose/imunologia , Suscetibilidade a Doenças , Haiti , Humanos , Imunidade Inata/genética , Imunidade Inata/fisiologia , Lactente , Lectina de Ligação a Manose/sangue , Lectina de Ligação a Manose/genética , Lectina de Ligação a Manose/imunologiaRESUMO
The seroepidemiology of Cryptosporidium infection was investigated in a representative sample of a normal population in the State of Sao Paulo, Brazil using a recombinant form of the immunodominant 27-kDa sporozoite antigen. IgG seropositivity was low in infants following loss of maternal antibody but quickly increased to approximately 60% by 5 years, then 80% by the age of 10 years, after which prevalence remained constant. The broad range of antibody concentrations is consistent with previous reports that the IgG response to C. parvum is short-lived. There is also evidence that average antibody concentrations increase with age. Results suggest that the recombinant antigen may be a more sensitive method of measuring seroprevalence than the native antigen in Western blot. Although cross-sectional studies can provide an insight into the epidemiology of C. parvum in normal populations, further studies investigating the dynamics of the humoral immune responses to Cryptosporidium and the use of serology in epidemiological studies are required.