The continued existence of New Zealand’s critically endangered and endemic black stilt or kakï (Himantopus novaezelandiae) relies on an intensive captive management programme. While this is successful at rearing large numbers of birds for release to the wild, poor survivability of these birds is limiting significant increases in the wild population. Predation and starvation are suspected to be the most common causes of death in released birds, but underlying contributing factors to these mortalities have not been fully evaluated. This research investigates the possible contribution of gastrointestinal (GI) helminth burdens and suboptimal digestive organ development to the high mortality rates of released kakï. Emphasis is placed on evaluating the methods used to assess the importance of these factors and to make informed recommendations for future management. The efficacy of the anthelmintic regime used for kakï was assessed by dosing half of the 80 captive birds with praziquantel (PZQ) prior to release in 2007. Faecal samples were collected before and after anthelmintic treatment, and before and after release to the wild. Post mortem worm counts were conducted on 11 birds that died following release and historical worm count records dating back to 1997 were accessed. The main findings were: PZQ had high efficacy against trematodes; treatment did not prevent re-infection; birds were exposed to helminths at release site; and there was no advantage of treatment for survival. Overall, the results suggest that anthelmintic treatment is an unnecessary cost. Consequently, recommendations were made to cease anthelmintic treatment or reduce its intensity, continue health screening, and incorporate annual efficacy testing to monitor the emergence of anthelmintic resistance. The reliability of faecal screening for GI helminths was evaluated. Faecal egg counts (FECs) were found to be poor indicators of worm burden. The two modified sedimentation methods used to detect trematodes provided relatively low egg recovery rates. Trematode egg shedding varied between days but not by hour of the day or temperature. The collection and analysis of pooled faecal samples proved to be more cost and time-effective than samples from individual birds and the larger masses collected resulted in greater sensitivity. In conclusion, faecal analysis of pooled samples is a useful qualitative indicator of helminth presence or absence but is quantitatively unreliable. In order to assess the importance of digestive organ development to captive-reared and released kak?, the digestive organs of healthy and emaciated captive, released and wild Himantopus sp. were compared. Captive and released kak? had generally smaller digestive organs than wild birds, released birds did not increase digestive organ size to match the high fibre wild diet, and emaciated birds did not have atrophied organs. However, the greatest mortality rates occur soon after release, while the birds were still being supplementary fed. These results suggest that reduced digestive efficiency is probably not contributing significantly to mortality rates and the direct impacts of the translocation are probably greater risk factors. The continued provision of supplementary food to released birds and an increased focus on habitat enhancement and predator control at release sites were recommended. The reliability of comparing fresh and formalin fixed Himantopus sp. gut specimens was evaluated. Significant differences in fresh and formalin fixed organ dry masses and variation in preserved organ lengths indicate that this variation should be considered in future studies. In conclusion, current management practices appear to be successful in ensuring that GI helminths and reduced digestive efficiency do not significantly lower the survivorship of captive-reared and released kakï. There is a need for further research into developing a more direct physiological assessment of the impacts of GI helminths and gut morphology as well as considering the role that starvation may have on poor survivability.

The continued existence of New Zealand’s critically endangered and endemic black stilt or kakï (Himantopus novaezelandiae) relies on an intensive captive management programme. While this is successful at rearing large numbers of birds for release to the wild, poor survivability of these birds is limiting significant increases in the wild population. Predation and starvation are suspected to be the most common causes of death in released birds, but underlying contributing factors to these mortalities have not been fully evaluated. This research investigates the possible contribution of gastrointestinal (GI) helminth burdens and suboptimal digestive organ development to the high mortality rates of released kakï. Emphasis is placed on evaluating the methods used to assess the importance of these factors and to make informed recommendations for future management. The efficacy of the anthelmintic regime used for kakï was assessed by dosing half of the 80 captive birds with praziquantel (PZQ) prior to release in 2007. Faecal samples were collected before and after anthelmintic treatment, and before and after release to the wild. Post mortem worm counts were conducted on 11 birds that died following release and historical worm count records dating back to 1997 were accessed. The main findings were: PZQ had high efficacy against trematodes; treatment did not prevent re-infection; birds were exposed to helminths at release site; and there was no advantage of treatment for survival. Overall, the results suggest that anthelmintic treatment is an unnecessary cost. Consequently, recommendations were made to cease anthelmintic treatment or reduce its intensity, continue health screening, and incorporate annual efficacy testing to monitor the emergence of anthelmintic resistance. The reliability of faecal screening for GI helminths was evaluated. Faecal egg counts (FECs) were found to be poor indicators of worm burden. The two modified sedimentation methods used to detect trematodes provided relatively low egg recovery rates. Trematode egg shedding varied between days but not by hour of the day or temperature. The collection and analysis of pooled faecal samples proved to be more cost and time-effective than samples from individual birds and the larger masses collected resulted in greater sensitivity. In conclusion, faecal analysis of pooled samples is a useful qualitative indicator of helminth presence or absence but is quantitatively unreliable. In order to assess the importance of digestive organ development to captive-reared and released kak?, the digestive organs of healthy and emaciated captive, released and wild Himantopus sp. were compared. Captive and released kak? had generally smaller digestive organs than wild birds, released birds did not increase digestive organ size to match the high fibre wild diet, and emaciated birds did not have atrophied organs. However, the greatest mortality rates occur soon after release, while the birds were still being supplementary fed. These results suggest that reduced digestive efficiency is probably not contributing significantly to mortality rates and the direct impacts of the translocation are probably greater risk factors. The continued provision of supplementary food to released birds and an increased focus on habitat enhancement and predator control at release sites were recommended. The reliability of comparing fresh and formalin fixed Himantopus sp. gut specimens was evaluated. Significant differences in fresh and formalin fixed organ dry masses and variation in preserved organ lengths indicate that this variation should be considered in future studies. In conclusion, current management practices appear to be successful in ensuring that GI helminths and reduced digestive efficiency do not significantly lower the survivorship of captive-reared and released kakï. There is a need for further research into developing a more direct physiological assessment of the impacts of GI helminths and gut morphology as well as considering the role that starvation may have on poor survivability.

The continued existence of New Zealand’s critically endangered and endemic black stilt or kakï (Himantopus novaezelandiae) relies on an intensive captive management programme. While this is successful at rearing large numbers of birds for release to the wild, poor survivability of these birds is limiting significant increases in the wild population. Predation and starvation are suspected to be the most common causes of death in released birds, but underlying contributing factors to these mortalities have not been fully evaluated. This research investigates the possible contribution of gastrointestinal (GI) helminth burdens and suboptimal digestive organ development to the high mortality rates of released kakï. Emphasis is placed on evaluating the methods used to assess the importance of these factors and to make informed recommendations for future management. The efficacy of the anthelmintic regime used for kakï was assessed by dosing half of the 80 captive birds with praziquantel (PZQ) prior to release in 2007. Faecal samples were collected before and after anthelmintic treatment, and before and after release to the wild. Post mortem worm counts were conducted on 11 birds that died following release and historical worm count records dating back to 1997 were accessed. The main findings were: PZQ had high efficacy against trematodes; treatment did not prevent re-infection; birds were exposed to helminths at release site; and there was no advantage of treatment for survival. Overall, the results suggest that anthelmintic treatment is an unnecessary cost. Consequently, recommendations were made to cease anthelmintic treatment or reduce its intensity, continue health screening, and incorporate annual efficacy testing to monitor the emergence of anthelmintic resistance. The reliability of faecal screening for GI helminths was evaluated. Faecal egg counts (FECs) were found to be poor indicators of worm burden. The two modified sedimentation methods used to detect trematodes provided relatively low egg recovery rates. Trematode egg shedding varied between days but not by hour of the day or temperature. The collection and analysis of pooled faecal samples proved to be more cost and time-effective than samples from individual birds and the larger masses collected resulted in greater sensitivity. In conclusion, faecal analysis of pooled samples is a useful qualitative indicator of helminth presence or absence but is quantitatively unreliable. In order to assess the importance of digestive organ development to captive-reared and released kak?, the digestive organs of healthy and emaciated captive, released and wild Himantopus sp. were compared. Captive and released kak? had generally smaller digestive organs than wild birds, released birds did not increase digestive organ size to match the high fibre wild diet, and emaciated birds did not have atrophied organs. However, the greatest mortality rates occur soon after release, while the birds were still being supplementary fed. These results suggest that reduced digestive efficiency is probably not contributing significantly to mortality rates and the direct impacts of the translocation are probably greater risk factors. The continued provision of supplementary food to released birds and an increased focus on habitat enhancement and predator control at release sites were recommended. The reliability of comparing fresh and formalin fixed Himantopus sp. gut specimens was evaluated. Significant differences in fresh and formalin fixed organ dry masses and variation in preserved organ lengths indicate that this variation should be considered in future studies. In conclusion, current management practices appear to be successful in ensuring that GI helminths and reduced digestive efficiency do not significantly lower the survivorship of captive-reared and released kakï. There is a need for further research into developing a more direct physiological assessment of the impacts of GI helminths and gut morphology as well as considering the role that starvation may have on poor survivability.

The continued existence of New Zealand’s critically endangered and endemic black stilt or kakï (Himantopus novaezelandiae) relies on an intensive captive management programme. While this is successful at rearing large numbers of birds for release to the wild, poor survivability of these birds is limiting significant increases in the wild population. Predation and starvation are suspected to be the most common causes of death in released birds, but underlying contributing factors to these mortalities have not been fully evaluated. This research investigates the possible contribution of gastrointestinal (GI) helminth burdens and suboptimal digestive organ development to the high mortality rates of released kakï. Emphasis is placed on evaluating the methods used to assess the importance of these factors and to make informed recommendations for future management. The efficacy of the anthelmintic regime used for kakï was assessed by dosing half of the 80 captive birds with praziquantel (PZQ) prior to release in 2007. Faecal samples were collected before and after anthelmintic treatment, and before and after release to the wild. Post mortem worm counts were conducted on 11 birds that died following release and historical worm count records dating back to 1997 were accessed. The main findings were: PZQ had high efficacy against trematodes; treatment did not prevent re-infection; birds were exposed to helminths at release site; and there was no advantage of treatment for survival. Overall, the results suggest that anthelmintic treatment is an unnecessary cost. Consequently, recommendations were made to cease anthelmintic treatment or reduce its intensity, continue health screening, and incorporate annual efficacy testing to monitor the emergence of anthelmintic resistance. The reliability of faecal screening for GI helminths was evaluated. Faecal egg counts (FECs) were found to be poor indicators of worm burden. The two modified sedimentation methods used to detect trematodes provided relatively low egg recovery rates. Trematode egg shedding varied between days but not by hour of the day or temperature. The collection and analysis of pooled faecal samples proved to be more cost and time-effective than samples from individual birds and the larger masses collected resulted in greater sensitivity. In conclusion, faecal analysis of pooled samples is a useful qualitative indicator of helminth presence or absence but is quantitatively unreliable. In order to assess the importance of digestive organ development to captive-reared and released kak?, the digestive organs of healthy and emaciated captive, released and wild Himantopus sp. were compared. Captive and released kak? had generally smaller digestive organs than wild birds, released birds did not increase digestive organ size to match the high fibre wild diet, and emaciated birds did not have atrophied organs. However, the greatest mortality rates occur soon after release, while the birds were still being supplementary fed. These results suggest that reduced digestive efficiency is probably not contributing significantly to mortality rates and the direct impacts of the translocation are probably greater risk factors. The continued provision of supplementary food to released birds and an increased focus on habitat enhancement and predator control at release sites were recommended. The reliability of comparing fresh and formalin fixed Himantopus sp. gut specimens was evaluated. Significant differences in fresh and formalin fixed organ dry masses and variation in preserved organ lengths indicate that this variation should be considered in future studies. In conclusion, current management practices appear to be successful in ensuring that GI helminths and reduced digestive efficiency do not significantly lower the survivorship of captive-reared and released kakï. There is a need for further research into developing a more direct physiological assessment of the impacts of GI helminths and gut morphology as well as considering the role that starvation may have on poor survivability.

The continued existence of New Zealand’s critically endangered and endemic black stilt or kakï (Himantopus novaezelandiae) relies on an intensive captive management programme. While this is successful at rearing large numbers of birds for release to the wild, poor survivability of these birds is limiting significant increases in the wild population. Predation and starvation are suspected to be the most common causes of death in released birds, but underlying contributing factors to these mortalities have not been fully evaluated. This research investigates the possible contribution of gastrointestinal (GI) helminth burdens and suboptimal digestive organ development to the high mortality rates of released kakï. Emphasis is placed on evaluating the methods used to assess the importance of these factors and to make informed recommendations for future management. The efficacy of the anthelmintic regime used for kakï was assessed by dosing half of the 80 captive birds with praziquantel (PZQ) prior to release in 2007. Faecal samples were collected before and after anthelmintic treatment, and before and after release to the wild. Post mortem worm counts were conducted on 11 birds that died following release and historical worm count records dating back to 1997 were accessed. The main findings were: PZQ had high efficacy against trematodes; treatment did not prevent re-infection; birds were exposed to helminths at release site; and there was no advantage of treatment for survival. Overall, the results suggest that anthelmintic treatment is an unnecessary cost. Consequently, recommendations were made to cease anthelmintic treatment or reduce its intensity, continue health screening, and incorporate annual efficacy testing to monitor the emergence of anthelmintic resistance. The reliability of faecal screening for GI helminths was evaluated. Faecal egg counts (FECs) were found to be poor indicators of worm burden. The two modified sedimentation methods used to detect trematodes provided relatively low egg recovery rates. Trematode egg shedding varied between days but not by hour of the day or temperature. The collection and analysis of pooled faecal samples proved to be more cost and time-effective than samples from individual birds and the larger masses collected resulted in greater sensitivity. In conclusion, faecal analysis of pooled samples is a useful qualitative indicator of helminth presence or absence but is quantitatively unreliable. In order to assess the importance of digestive organ development to captive-reared and released kak?, the digestive organs of healthy and emaciated captive, released and wild Himantopus sp. were compared. Captive and released kak? had generally smaller digestive organs than wild birds, released birds did not increase digestive organ size to match the high fibre wild diet, and emaciated birds did not have atrophied organs. However, the greatest mortality rates occur soon after release, while the birds were still being supplementary fed. These results suggest that reduced digestive efficiency is probably not contributing significantly to mortality rates and the direct impacts of the translocation are probably greater risk factors. The continued provision of supplementary food to released birds and an increased focus on habitat enhancement and predator control at release sites were recommended. The reliability of comparing fresh and formalin fixed Himantopus sp. gut specimens was evaluated. Significant differences in fresh and formalin fixed organ dry masses and variation in preserved organ lengths indicate that this variation should be considered in future studies. In conclusion, current management practices appear to be successful in ensuring that GI helminths and reduced digestive efficiency do not significantly lower the survivorship of captive-reared and released kakï. There is a need for further research into developing a more direct physiological assessment of the impacts of GI helminths and gut morphology as well as considering the role that starvation may have on poor survivability.

The continued existence of New Zealand’s critically endangered and endemic black stilt or kakï (Himantopus novaezelandiae) relies on an intensive captive management programme. While this is successful at rearing large numbers of birds for release to the wild, poor survivability of these birds is limiting significant increases in the wild population. Predation and starvation are suspected to be the most common causes of death in released birds, but underlying contributing factors to these mortalities have not been fully evaluated. This research investigates the possible contribution of gastrointestinal (GI) helminth burdens and suboptimal digestive organ development to the high mortality rates of released kakï. Emphasis is placed on evaluating the methods used to assess the importance of these factors and to make informed recommendations for future management. The efficacy of the anthelmintic regime used for kakï was assessed by dosing half of the 80 captive birds with praziquantel (PZQ) prior to release in 2007. Faecal samples were collected before and after anthelmintic treatment, and before and after release to the wild. Post mortem worm counts were conducted on 11 birds that died following release and historical worm count records dating back to 1997 were accessed. The main findings were: PZQ had high efficacy against trematodes; treatment did not prevent re-infection; birds were exposed to helminths at release site; and there was no advantage of treatment for survival. Overall, the results suggest that anthelmintic treatment is an unnecessary cost. Consequently, recommendations were made to cease anthelmintic treatment or reduce its intensity, continue health screening, and incorporate annual efficacy testing to monitor the emergence of anthelmintic resistance. The reliability of faecal screening for GI helminths was evaluated. Faecal egg counts (FECs) were found to be poor indicators of worm burden. The two modified sedimentation methods used to detect trematodes provided relatively low egg recovery rates. Trematode egg shedding varied between days but not by hour of the day or temperature. The collection and analysis of pooled faecal samples proved to be more cost and time-effective than samples from individual birds and the larger masses collected resulted in greater sensitivity. In conclusion, faecal analysis of pooled samples is a useful qualitative indicator of helminth presence or absence but is quantitatively unreliable. In order to assess the importance of digestive organ development to captive-reared and released kak?, the digestive organs of healthy and emaciated captive, released and wild Himantopus sp. were compared. Captive and released kak? had generally smaller digestive organs than wild birds, released birds did not increase digestive organ size to match the high fibre wild diet, and emaciated birds did not have atrophied organs. However, the greatest mortality rates occur soon after release, while the birds were still being supplementary fed. These results suggest that reduced digestive efficiency is probably not contributing significantly to mortality rates and the direct impacts of the translocation are probably greater risk factors. The continued provision of supplementary food to released birds and an increased focus on habitat enhancement and predator control at release sites were recommended. The reliability of comparing fresh and formalin fixed Himantopus sp. gut specimens was evaluated. Significant differences in fresh and formalin fixed organ dry masses and variation in preserved organ lengths indicate that this variation should be considered in future studies. In conclusion, current management practices appear to be successful in ensuring that GI helminths and reduced digestive efficiency do not significantly lower the survivorship of captive-reared and released kakï. There is a need for further research into developing a more direct physiological assessment of the impacts of GI helminths and gut morphology as well as considering the role that starvation may have on poor survivability.

The continued existence of New Zealand’s critically endangered and endemic black stilt or kakï (Himantopus novaezelandiae) relies on an intensive captive management programme. While this is successful at rearing large numbers of birds for release to the wild, poor survivability of these birds is limiting significant increases in the wild population. Predation and starvation are suspected to be the most common causes of death in released birds, but underlying contributing factors to these mortalities have not been fully evaluated. This research investigates the possible contribution of gastrointestinal (GI) helminth burdens and suboptimal digestive organ development to the high mortality rates of released kakï. Emphasis is placed on evaluating the methods used to assess the importance of these factors and to make informed recommendations for future management. The efficacy of the anthelmintic regime used for kakï was assessed by dosing half of the 80 captive birds with praziquantel (PZQ) prior to release in 2007. Faecal samples were collected before and after anthelmintic treatment, and before and after release to the wild. Post mortem worm counts were conducted on 11 birds that died following release and historical worm count records dating back to 1997 were accessed. The main findings were: PZQ had high efficacy against trematodes; treatment did not prevent re-infection; birds were exposed to helminths at release site; and there was no advantage of treatment for survival. Overall, the results suggest that anthelmintic treatment is an unnecessary cost. Consequently, recommendations were made to cease anthelmintic treatment or reduce its intensity, continue health screening, and incorporate annual efficacy testing to monitor the emergence of anthelmintic resistance. The reliability of faecal screening for GI helminths was evaluated. Faecal egg counts (FECs) were found to be poor indicators of worm burden. The two modified sedimentation methods used to detect trematodes provided relatively low egg recovery rates. Trematode egg shedding varied between days but not by hour of the day or temperature. The collection and analysis of pooled faecal samples proved to be more cost and time-effective than samples from individual birds and the larger masses collected resulted in greater sensitivity. In conclusion, faecal analysis of pooled samples is a useful qualitative indicator of helminth presence or absence but is quantitatively unreliable. In order to assess the importance of digestive organ development to captive-reared and released kak?, the digestive organs of healthy and emaciated captive, released and wild Himantopus sp. were compared. Captive and released kak? had generally smaller digestive organs than wild birds, released birds did not increase digestive organ size to match the high fibre wild diet, and emaciated birds did not have atrophied organs. However, the greatest mortality rates occur soon after release, while the birds were still being supplementary fed. These results suggest that reduced digestive efficiency is probably not contributing significantly to mortality rates and the direct impacts of the translocation are probably greater risk factors. The continued provision of supplementary food to released birds and an increased focus on habitat enhancement and predator control at release sites were recommended. The reliability of comparing fresh and formalin fixed Himantopus sp. gut specimens was evaluated. Significant differences in fresh and formalin fixed organ dry masses and variation in preserved organ lengths indicate that this variation should be considered in future studies. In conclusion, current management practices appear to be successful in ensuring that GI helminths and reduced digestive efficiency do not significantly lower the survivorship of captive-reared and released kakï. There is a need for further research into developing a more direct physiological assessment of the impacts of GI helminths and gut morphology as well as considering the role that starvation may have on poor survivability.

Diss. (sammanfattning) Umeå : Univ., 2009. / Härtill 4 uppsatser.

Made available in DSpace on 2016-12-23T13:56:03Z (GMT). No. of bitstreams: 1 HELMINtiASE INTESTINAL.pdf: 1507224 bytes, checksum: 6132aca0f1aadf166d4e2b773c982a74 (MD5) Previous issue date: 2009-04-27 / Mycobacterium tuberculosis (MTB) é um exemplo clássico de patógeno para o qual a resposta protetora depende da imunidade celular do tipo Th1, que é caracterizada pela presença de linfócitos T CD4+ produtores de IFN-g. Essa citocina ativa mecanismos microbicidas no macrófago infectado, levando à eliminação do bacilo. Evidências sugerem que a progressão para a tuberculose esteja relacionada à presença de mecanismos imunossupressores mediados por citocinas e por células T reguladoras. Acredita-se que a presença de helmintíase intestinal possa prejudicar o desenvolvimento de uma resposta adaptativa capaz de conter ou eliminar o MTB, tornando assim o indivíduo susceptível ao adoecimento. Para aquilatar a influência da infecção por helmintos intestinais na resposta celular durante a tuberculose pulmonar, neste trabalho, foram avaliados os perfis quantitativo e fenotípico de populações celulares de sangue periférico e o padrão de citocinas em culturas de sangue total estimuladas com antígenos de MTB, em pacientes portadores de tuberculose pulmonar apresentando ou não helmintíase intestinal no momento do diagnóstico e durante a terapia antituberculose. Para isso, foram arrolados 53 pacientes com diagnóstico recente de tuberculose pulmonar. Desses, 26% eram portadores de pelo menos uma espécie de helminto intestinal. Pacientes com tuberculose pulmonar apresentaram uma redução significativa nos números de linfócitos T CD8+, células NK e NKT. Os indivíduos com helmintíase intestinal associada à tuberculose apresentaram uma maior freqüência de células T reguladoras, com ambos os fenótipos CD4+CD25HIGH e CD4+CD25HIGHFoxp3+. Além disso, os resultados sugerem que a presença de infecção por helmintos intestinais tenha induzido um estado de hipoergia em pacientes portadores de tuberculose pulmonar, uma vez que esses pacientes apresentaram concentrações menores das citocinas IL-2, TNF-a, IL-4, IL-5 e IL-10 nos sobrenadantes de culturas em relação às concentrações encontradas no grupo TB e no grupo controle. Portanto, os resultados desse trabalho sugerem que a presença de infecção por helmintos intestinais tenha um impacto negativo na resposta imunitária à tuberculose em pacientes portadores de tuberculose pulmonar. / The protection against Mycobacterium tuberculosis (MTB) depends on a cellmediated Th1 type-immune response. This response is characterized by IFN-g production by CD4+ T cells, which activates macrophages to enhance microbicidal mechanisms leading to the bacillus eradication. Factors related to tuberculosis resistance or susceptibility are not completely understood. There are evidences suggesting that the progress to active disease is related to immune downregulation caused by suppressors cytokines and regulatory T cells. It is believed that the association with helminth infection can disturb the protective immune response that should contain or eliminate MTB. Here, we investigated the role of intestinal helminth infection on M. tuberculosis specific immune response during active pulmonar tuberculosis in patients with associated tuberculosis and intestinal helminth infection at the time of diagnosis and during tuberculosis therapy. Quantitative and phenotypic analyses of peripheral blood cells populations were performed and the MTBstimulated whole blood culture cytokines production was evaluated. Fifty-three patients with newly diagnosed tuberculosis were enrolled for this study. Twenty-six percent of these patients were infected with at least one intestinal helminth (TB + HELM patients). Patients with pulmonary tuberculosis presented a significant reduction in the numbers of TCD8+, NK and NKT cells. Patients with both intestinal helminth infection and tuberculosis presented higher frequency of regulatory T cells, of both phenotype CD4+CD25HIGH and CD4+CD25HIGHFoxp3+, as compared to TB group, to HELM group, and to control group. In addition, the results suggest a hipoergy status in TB + HELM patients because the production of the cytokines IL-2, TNF-a, IL-4, IL-5 and IL-10 decreased in whole blood culture of these patients as compared to both TB patients and healthy controls. The data from this study indicated that the associated intestinal helminth infection has a negative impact on immunity to tuberculosis in patients with tuberculosis.

A estrongiloidíase é a infecção parasitária causada pelo nematódeo Strongyloides stercoralis. O diagnóstico definitivo é realizado pela visualização de larvas, principalmente nas fezes. Porém as técnicas parasitológicas têm baixa sensibilidade. As técnicas sorológicas apresentam-se como importante alternativa diagnóstica. Pesquisas apontam para a utilização de antígenos heterólogos solúveis, principalmente de Strongyloides venezuelensis. A identificação e caracterização dos antígenos de membrana podem fornecer fonte alternativa de antígenos e assim auxiliar o desenvolvimento das técnicas imunológicas. O presente trabalho teve como objetivo a avaliação das técnicas ELISA e WB frente a diferentes frações antigênicas de larvas filarioides de S. venezuelensis. Foram utilizadas amostras de sangue e fezes de 92 indivíduos, 20 indivíduos com estrongiloidíase (grupo I), 32 indivíduos com outras parasitoses (grupo II) e 40 indivíduos negativos (grupo III) pelos métodos de Lutz, cultura em placa de ágar e Rugai. Para preparação dos antígenos foram utilizadas larvas infectantes obtidas a partir de ratos infectados experimentalmente com S. venezuelensis. Seis frações antigênicas foram preparadas: frações salinas solúveis e de membrana (PBS 0,01M pH 7,2 e SDS 1%, SS e MS; Tris-HCl 25mM pH 7,5 e CHAPS 1%, ST e MT, respectivamente) e frações alcalinas solúvel e de membrana (NaOH 0,15M e SDS 1%, SA e MA, respectivamente). Para a técnica ELISA foram utilizadas placas sensibilizadas com 10ug/mL de antígeno, soro dos indivíduos diluídos 1:200 em PBS 0,05% Tween 3% de leite (PBSTM) e o conjugado (anti IgG-humana peroxidase) em PBSTM. As amostras foram consideradas positivas quando o Índice ELISA foi maior que 1. Para a técnica de WB os soros foram diluídos 1:100 em Tris-HCl 5% de leite (TM) e o conjugado (anti IgG-humana peroxidase) em TM. Após as técnicas sorológicas foram determinadas os parâmetros de diagnóstico pela curva ROC como sensibilidade (SE), especificidade (ES), Likelihood ratio (LR) além da determinação da acurácia diagnóstica (AC) e do índice Kappa (k). A técnica ELISA destacou as frações de membrana com melhor desempenho em relação aos parêmetros diagnósticos estudados (SE 95%, ES 94,4%, AC 94,8%, LR 17,1, k 0,848). O WB revelou componentes antigênicos imunodominantes variando de 260-10kDa, mas destacam-se as frações de 40-35kDa mais frequentes em todas frações antigênicas. Pela técnica de WB, a fração ST apresentou melhor desempenho em relação aos parêmetros diagnósticos estudados (SE 100%, ES 93,1%, AC 94,5, LR 14,4,k 0,854). A utilização das frações de membrana no imunodiagnóstico da estrongiloidíase humana torna-se fonte acessível e eficaz em relação às frações purificadas, não necessitando de gastos complementares para sua obtenção / Strongyloidiasis is a parasitic infection caused by a nematode Strongyloides stercoralis. The definitive diagnosis is made by the larvae visualization in stool samples. However parasitological techniques have low sensitivity. Serological techniques became as suitable diagnostic alternative. Research indicates for the soluble heterologous antigen utilization, mainly Strongyloides venezuelensis. Identification and characterization of membrane antigen may constitute an alternative source of antigen and then assist the development of serological techniques. The aim of this study was evaluate ELISA and WB techniques behind different antigenic fractions of S. venezuelensis´ infective larvae. A total of 92 serum and stool samples was analyzed, 20 from individuals with strongyloidisis (group 1), 32 with other parasitic diseases (group 2) and 40 from individuals with negative coproparasitology (group 3) using Lutz, agar plate culture and Rugai methods. For the antigen preparation infective larvae of S. venezuelensis from experimental infected rats were employed. Six antigenic fractions were prepareted: saline soluble and from membrane fractions (0.01M PBS pH 7.2, and 1% SDS, SS and MS; 25mM Tris-HCl pH 7.5, 1% CHAPS, MT and ST, respectively) and alkaline soluble and membrane fractions (0.15 M NaOH and 1% SDS, SA and MA, respectively). For ELISA technique, plates were sensitized with 10 ug/mL of antigen, serum samples were diluted 1:200 in 0.05% Tween in PBS 3% milk (PBSTM) and conjugate (anti-human IgG peroxidase) in PBSTM. Positive samples were considered when ELISA index was greater than 1. To WB technique, serum samples were diluted 1:100 in Tris-HCl 5% milk (TM) and conjugate (anti-human IgG peroxidase) in the TM. After serological techniques diagnostics parameters were determined by ROC curve how sensitivity (SE), specificity (ES), Likelihood ratio (LR) and determination of diagnostic accuracy (AC) and Kappa (k) index. ELISA technique highlighted the membrane fractions with better performance compared to parameters diagnoses studied (95% SE, 94.4% ES, 94.8% AC, 17.1 LR, 0.848 k). The WB revealed immunodominant antigenic components ranging from 260-10kDa, but there are the fractions of 40-35kDa more frequent in all antigenic fractions. WB technique showed ST fraction better performance in relation to the diagnostic parameters (100% SE, 93.1% ES, 94.5% AC, 14.4 LR, 0.854 k). Membrane fractions in the immunodiagnosis of human strongyloidiasis become an accessible and effective source of antigens in relation to the purified fractions, requiring no additional expense to obtain it