Made available in DSpace on 2014-06-11T19:32:51Z (GMT). No. of bitstreams: 0 Previous issue date: 2011-08-25Bitstream added on 2014-06-13T18:44:23Z : No. of bitstreams: 1 coelho_wmd_dr_jabo.pdf: 4287442 bytes, checksum: 7091394609cf026e1187fcfeac0e61e9 (MD5) / Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP) / A criptosporidiose é uma doença entérica grave, economicamente significante, caracterizada principalmente por desordens intestinais, podendo ocasionar manifestações clínicas variadas e eventual mortalidade, principalmente em animais jovens. Este estudo objetivou detectar molecularmente genótipos e subgenótipos de Cryptosporidium spp. em fezes de cabritos provenientes dos Estados de Goiás, Mato Grosso do Sul, Minas Gerais e São Paulo, Brasil. Amostras fecais foram colhidas diretamente do reto de 192 cabritos de diferentes raças, machos e fêmeas, com até doze meses de idade. Adicionalmente, foram analisadas amostras fecais de ovinos, bovinos, suínos e eqüinos jovens. A eliminação de oocistos de Cryptosporidium spp. foi observada por meio das técnicas de Sheather e Kinyoun, seguindo-se a micrometragem dos oocistos com ocular de campo amplo micrométrica 10x (Bioval®) em aumento microscópico de 400 e 1000x. A reação em cadeia da polimerase (PCR) foi realizada para amplificação dos fragmentos dos genes da subunidade 18S do rRNA e da glicoproteína GP60. Presença de oocistos de Cryptosporidium spp. foram observados pela análise microscópica em 11,45% (22/192) das amostras analisadas. Amplificação gênica positiva para Cryptosporidium foi demonstrada em 16,66% (32/192) destas amostras. Com o sequenciamento dos produtos da PCR do gene 18S rRNA, todas as amostras foram identificadas como Cryptosporidium parvum. Por meio da subgenotipagem com o sequenciamento do gene GP60, foi encontrado exclusivamente o subgenótipo de C. parvum IIaA15G2R1. Através dos resultados obtidos, pode-se inferir que a infecção por C. parvum está presente em rebanhos caprinos de diferentes Estados brasileiros podendo, esta espécie animal, atuar como uma importante fonte de infecção do subtipo zoonótico de Cryptosporidium para outras espécies animais, em especial para o ser humano / Cryptosporidiosis is a serious enteric disease, economically significant, mainly characterized by intestinal disorders, may cause various clinical manifestations and eventual mortality, especially in young animals. This study aimed to detect and molecularly genotypes and subgenotypes of Cryptosporidium spp. in feces of goat kids from the states of Goiás, Mato Grosso do Sul, Minas Gerais and São Paulo, Brazil. Fecal samples were collected directly from the rectum of 192 goat kids of different breeds, males and females, with up to twelve months old. Additionally, were analyzed fecal samples from cattle, sheep, pigs and young horses. The elimination of oocysts of Cryptosporidium spp. was observed using the Sheather and Kinyoun techniques, followed by micrometric analysis with ocular micrometer wide-field 10x (Bioval ®) in increase from 400 and 1000x. The polymerase chain reaction (PCR) was performed to amplify fragments of genes of the subunit 18S rRNA and glycoprotein GP60. Presence of Cryptosporidium spp. was observed by microscopic examination in 11.45% (22/192) of the samples. Gene amplification for Cryptosporidium was demonstrated in 16.66% (32/192) of these samples. With the sequencing of the PCR products of 18S rRNA gene, all samples were identified as Cryptosporidium parvum. Through of subgenotyping with sequencing of GP60 gene was found exclusively the subtype of C. parvum IIaA15G2R1. By the results obtained, it can be inferred that infection with C. parvum is present in goat kids in different brazilian states may, this animal species act as an important source of infection with zoonotic subtype of Cryptosporidium to other animal species, especially for humans

Orientador: Katia Denise Saraiva Bresciani / Coorientador: Marcelo Vasconcelos Meireles / Banca: Luiz Eduardo Corrêa Fonseca / Banca: Adjair Antonio do Nascimento / Banca: Jancarlo Ferreira Gomes / Banca: Luiz Augusto do Amaral / Resumo: A criptosporidiose é uma doença entérica grave, economicamente significante, caracterizada principalmente por desordens intestinais, podendo ocasionar manifestações clínicas variadas e eventual mortalidade, principalmente em animais jovens. Este estudo objetivou detectar molecularmente genótipos e subgenótipos de Cryptosporidium spp. em fezes de cabritos provenientes dos Estados de Goiás, Mato Grosso do Sul, Minas Gerais e São Paulo, Brasil. Amostras fecais foram colhidas diretamente do reto de 192 cabritos de diferentes raças, machos e fêmeas, com até doze meses de idade. Adicionalmente, foram analisadas amostras fecais de ovinos, bovinos, suínos e eqüinos jovens. A eliminação de oocistos de Cryptosporidium spp. foi observada por meio das técnicas de Sheather e Kinyoun, seguindo-se a micrometragem dos oocistos com ocular de campo amplo micrométrica 10x (Bioval®) em aumento microscópico de 400 e 1000x. A reação em cadeia da polimerase (PCR) foi realizada para amplificação dos fragmentos dos genes da subunidade 18S do rRNA e da glicoproteína GP60. Presença de oocistos de Cryptosporidium spp. foram observados pela análise microscópica em 11,45% (22/192) das amostras analisadas. Amplificação gênica positiva para Cryptosporidium foi demonstrada em 16,66% (32/192) destas amostras. Com o sequenciamento dos produtos da PCR do gene 18S rRNA, todas as amostras foram identificadas como Cryptosporidium parvum. Por meio da subgenotipagem com o sequenciamento do gene GP60, foi encontrado exclusivamente o subgenótipo de C. parvum IIaA15G2R1. Através dos resultados obtidos, pode-se inferir que a infecção por C. parvum está presente em rebanhos caprinos de diferentes Estados brasileiros podendo, esta espécie animal, atuar como uma importante fonte de infecção do subtipo zoonótico de Cryptosporidium para outras espécies animais, em especial para o ser humano / Abstract: Cryptosporidiosis is a serious enteric disease, economically significant, mainly characterized by intestinal disorders, may cause various clinical manifestations and eventual mortality, especially in young animals. This study aimed to detect and molecularly genotypes and subgenotypes of Cryptosporidium spp. in feces of goat kids from the states of Goiás, Mato Grosso do Sul, Minas Gerais and São Paulo, Brazil. Fecal samples were collected directly from the rectum of 192 goat kids of different breeds, males and females, with up to twelve months old. Additionally, were analyzed fecal samples from cattle, sheep, pigs and young horses. The elimination of oocysts of Cryptosporidium spp. was observed using the Sheather and Kinyoun techniques, followed by micrometric analysis with ocular micrometer wide-field 10x (Bioval ®) in increase from 400 and 1000x. The polymerase chain reaction (PCR) was performed to amplify fragments of genes of the subunit 18S rRNA and glycoprotein GP60. Presence of Cryptosporidium spp. was observed by microscopic examination in 11.45% (22/192) of the samples. Gene amplification for Cryptosporidium was demonstrated in 16.66% (32/192) of these samples. With the sequencing of the PCR products of 18S rRNA gene, all samples were identified as Cryptosporidium parvum. Through of subgenotyping with sequencing of GP60 gene was found exclusively the subtype of C. parvum IIaA15G2R1. By the results obtained, it can be inferred that infection with C. parvum is present in goat kids in different brazilian states may, this animal species act as an important source of infection with zoonotic subtype of Cryptosporidium to other animal species, especially for humans / Doutor

An enhanced notified enteric disease surveillance trial began on 1 July 2007 and continued until 30 June 2008. The aim of the trial was to measure the quality, timeliness and completeness of data collected and submitted by a regional Public Health Service (PHS) to the Institute of Environmental Science and Research Limited (ESR), via the national disease database (EpiSurv) for notified cases of enteric diseases. The trial evaluated two different methods of data collection: postal questionnaires and telephone interviews. Telephone interview techniques were used to improve the contact rate, timeliness and completeness of data gathered from all notified cases of campylobacteriosis in the Manawatu, Horowhenua and Tararua regions. The target set for the project was to achieve a 95% contact rate with 90% full completion of all EpiSurv data fields. For all notified cases of campylobacteriosis a 97% contact rate was achieved in a time frame of between zero to 20 days (three day median) and completeness of all the EpiSurv case report fields ranged between 96 – 100% in the final data. Prior to the commencement of the study, between 1 July 2004 to 30 June 2005, MidCentral PHS (MCPHS) made contact with around 58% of all notified cases of campylobacteriosis and 77% of all other notified enteric disease cases1 . A short pre-screen mail questionnaire, with reply-paid envelope, was sent to all notified cases of cryptosporidiosis, giardiasis, salmonellosis and yersiniosis in the MCPHS regions. EpiSurv case report fields were completed using information supplied in the returned questionnaires. Return rate, timeliness, and completeness were compared with the telephone interview group. Fifty three percent of cases we attempted to contact via mail questionnaire responded within two to 63 days (six day median) and completeness of all the EpiSurv case report fields ranged between 81 – 100%. In addition, we monitored the newly introduced ESR Early Aberration Reporting System (EARS) flags for increased levels of disease compared to historical disease rates, and assessed its usefulness as a tool to identify potential outbreaks in the region. While no outbreaks that had not already been identified by PHS staff were found by monitoring the EARS system, EARS has become an important tool in the MCPHS for comparing our rates of disease with bordering PHSs. EARS also provided a good quick reference tool for media enquiries and the graphs produced in EARS have been well utilised as visual aids for training and seminars presented during the trial period. The results of the surveillance trial initiatives were compared to the rest of New Zealand (NZ) over the same time frame and with a comparable, medium-sized, PHS. While the results of the telephone interviews from the MCPHS trial were close to the comparable PHS, they were significantly higher than for the rest of NZ. The postal questionnaires achieved a lower contact rate than the comparable PHS but similar to the rest of NZ. However, the quality of data gathered in the returned MCPHS postal questionnaire was significantly higher in most fields. Additional analysis was undertaken which indicated that those cases living in higher deprivation and rural areas were less likely to respond to a postal questionnaire. An over-representation of common enteric disease notifications from rural areas in the MCPHS was also highlighted by our research. This trial has shown the effectiveness of utilising telephone interviews and telemarketing techniques for gathering timely and complete data for human enteric disease surveillance within the MCPHS. It has also demonstrated that a short pre-screen questionnaire can be effective in collecting good quality data needed to complete the standard EpiSurv case report form.

An enhanced notified enteric disease surveillance trial began on 1 July 2007 and continued until 30 June 2008. The aim of the trial was to measure the quality, timeliness and completeness of data collected and submitted by a regional Public Health Service (PHS) to the Institute of Environmental Science and Research Limited (ESR), via the national disease database (EpiSurv) for notified cases of enteric diseases. The trial evaluated two different methods of data collection: postal questionnaires and telephone interviews. Telephone interview techniques were used to improve the contact rate, timeliness and completeness of data gathered from all notified cases of campylobacteriosis in the Manawatu, Horowhenua and Tararua regions. The target set for the project was to achieve a 95% contact rate with 90% full completion of all EpiSurv data fields. For all notified cases of campylobacteriosis a 97% contact rate was achieved in a time frame of between zero to 20 days (three day median) and completeness of all the EpiSurv case report fields ranged between 96 – 100% in the final data. Prior to the commencement of the study, between 1 July 2004 to 30 June 2005, MidCentral PHS (MCPHS) made contact with around 58% of all notified cases of campylobacteriosis and 77% of all other notified enteric disease cases1 . A short pre-screen mail questionnaire, with reply-paid envelope, was sent to all notified cases of cryptosporidiosis, giardiasis, salmonellosis and yersiniosis in the MCPHS regions. EpiSurv case report fields were completed using information supplied in the returned questionnaires. Return rate, timeliness, and completeness were compared with the telephone interview group. Fifty three percent of cases we attempted to contact via mail questionnaire responded within two to 63 days (six day median) and completeness of all the EpiSurv case report fields ranged between 81 – 100%. In addition, we monitored the newly introduced ESR Early Aberration Reporting System (EARS) flags for increased levels of disease compared to historical disease rates, and assessed its usefulness as a tool to identify potential outbreaks in the region. While no outbreaks that had not already been identified by PHS staff were found by monitoring the EARS system, EARS has become an important tool in the MCPHS for comparing our rates of disease with bordering PHSs. EARS also provided a good quick reference tool for media enquiries and the graphs produced in EARS have been well utilised as visual aids for training and seminars presented during the trial period. The results of the surveillance trial initiatives were compared to the rest of New Zealand (NZ) over the same time frame and with a comparable, medium-sized, PHS. While the results of the telephone interviews from the MCPHS trial were close to the comparable PHS, they were significantly higher than for the rest of NZ. The postal questionnaires achieved a lower contact rate than the comparable PHS but similar to the rest of NZ. However, the quality of data gathered in the returned MCPHS postal questionnaire was significantly higher in most fields. Additional analysis was undertaken which indicated that those cases living in higher deprivation and rural areas were less likely to respond to a postal questionnaire. An over-representation of common enteric disease notifications from rural areas in the MCPHS was also highlighted by our research. This trial has shown the effectiveness of utilising telephone interviews and telemarketing techniques for gathering timely and complete data for human enteric disease surveillance within the MCPHS. It has also demonstrated that a short pre-screen questionnaire can be effective in collecting good quality data needed to complete the standard EpiSurv case report form.

An enhanced notified enteric disease surveillance trial began on 1 July 2007 and continued until 30 June 2008. The aim of the trial was to measure the quality, timeliness and completeness of data collected and submitted by a regional Public Health Service (PHS) to the Institute of Environmental Science and Research Limited (ESR), via the national disease database (EpiSurv) for notified cases of enteric diseases. The trial evaluated two different methods of data collection: postal questionnaires and telephone interviews. Telephone interview techniques were used to improve the contact rate, timeliness and completeness of data gathered from all notified cases of campylobacteriosis in the Manawatu, Horowhenua and Tararua regions. The target set for the project was to achieve a 95% contact rate with 90% full completion of all EpiSurv data fields. For all notified cases of campylobacteriosis a 97% contact rate was achieved in a time frame of between zero to 20 days (three day median) and completeness of all the EpiSurv case report fields ranged between 96 – 100% in the final data. Prior to the commencement of the study, between 1 July 2004 to 30 June 2005, MidCentral PHS (MCPHS) made contact with around 58% of all notified cases of campylobacteriosis and 77% of all other notified enteric disease cases1 . A short pre-screen mail questionnaire, with reply-paid envelope, was sent to all notified cases of cryptosporidiosis, giardiasis, salmonellosis and yersiniosis in the MCPHS regions. EpiSurv case report fields were completed using information supplied in the returned questionnaires. Return rate, timeliness, and completeness were compared with the telephone interview group. Fifty three percent of cases we attempted to contact via mail questionnaire responded within two to 63 days (six day median) and completeness of all the EpiSurv case report fields ranged between 81 – 100%. In addition, we monitored the newly introduced ESR Early Aberration Reporting System (EARS) flags for increased levels of disease compared to historical disease rates, and assessed its usefulness as a tool to identify potential outbreaks in the region. While no outbreaks that had not already been identified by PHS staff were found by monitoring the EARS system, EARS has become an important tool in the MCPHS for comparing our rates of disease with bordering PHSs. EARS also provided a good quick reference tool for media enquiries and the graphs produced in EARS have been well utilised as visual aids for training and seminars presented during the trial period. The results of the surveillance trial initiatives were compared to the rest of New Zealand (NZ) over the same time frame and with a comparable, medium-sized, PHS. While the results of the telephone interviews from the MCPHS trial were close to the comparable PHS, they were significantly higher than for the rest of NZ. The postal questionnaires achieved a lower contact rate than the comparable PHS but similar to the rest of NZ. However, the quality of data gathered in the returned MCPHS postal questionnaire was significantly higher in most fields. Additional analysis was undertaken which indicated that those cases living in higher deprivation and rural areas were less likely to respond to a postal questionnaire. An over-representation of common enteric disease notifications from rural areas in the MCPHS was also highlighted by our research. This trial has shown the effectiveness of utilising telephone interviews and telemarketing techniques for gathering timely and complete data for human enteric disease surveillance within the MCPHS. It has also demonstrated that a short pre-screen questionnaire can be effective in collecting good quality data needed to complete the standard EpiSurv case report form.