RESUMO
Recent reports of hookworm infection in Alabama, USA, has prompted surveillance in Mississippi, given the states' similar environmental conditions. We collected stool specimens from 277 children in Rankin County, Mississippi. Kato-Katz microscopic smear, agar plate culture, and quantitative PCR indicated no soil-transmitted helminths. Nevertheless, further surveillance in other high-risk Mississippi counties is warranted.
Assuntos
Helmintos , Solo , Criança , Animais , Humanos , Solo/parasitologia , Mississippi/epidemiologia , Fezes/parasitologia , Prevalência , Helmintos/genéticaRESUMO
Hookworm infection is endemic in many countries throughout the world; however, the information about the prevalence of each species, Necatoramericanus and Ancylostomaduodenale, is inaccurate in many South American countries. We aimed to determine the prevalence of human hookworm species by combining the results of both microscopy and PCR among endemic populations in Argentina, represented by natives and immigrants. A total of 140 serial fecal specimens were obtained from natives in the province of Misiones and an immigrant community living in the province of Buenos Aires. Samples were examined using the formalin-ethyl acetate concentration technique (FECT) and one flotation technique (screening tests) and specific PCRs for N. americanus and A. duodenale. We characterized samples containing N. americanus by sequencing a fragment of the cytochrome b gene. The observed hookworm prevalence as assessed by the screening tests and PCR were 24.3% and 32.8%, respectively. PCR positive samples were identified as N. americanus. PCR had 100% sensitivity compared with 73.9% of screening tests. A total of 12 samples from individuals with hookworm-infected household members were positive only by PCR. N. americanus sequences showed 90.5% identity, being more similar to each other than to any of the sequences obtained from GenBank. This is the first study that provides molecular data and characterization of N. americanus in Argentina. The complementary use of FECT and one flotation technique to screen hookworm infections, followed by PCR to differentiate the species contribute to produce better prevalence estimates.
Assuntos
Ancylostoma , Necator americanus , Animais , Humanos , Ancylostoma/genética , Necator americanus/genética , Ancylostomatoidea , Diagnóstico Diferencial , Argentina/epidemiologia , FezesRESUMO
Canine parvovirus type 2 (CPV-2) is among the most important and highly contagious pathogens that cause enteric or systemic infections in domestic and nondomestic carnivores. However, the spillover of CPV-2 to noncarnivores is rarely mentioned. Taiwanese pangolins (Manis pentadactyla pentadactyla) are threatened due to habitat fragmentation and prevalent animal trafficking. Interactions between Taiwanese pangolins, humans, and domestic animals have become more frequent in recent years. However, information about the susceptibility of pangolins to common infectious agents of domestic animals has been lacking. From October 2017 to June 2019, 4 pangolins that were rescued and treated in wildlife rescue centers in central and northern Taiwan presented with gastrointestinal signs. Gross and histopathological examination revealed the main pathologic changes to be necrotic enteritis with involvement of the crypts in all intestinal segments in 2 pangolins. By immunohistochemistry for CPV-2, there was positive labeling of cryptal epithelium throughout the intestine, and immunolabeling was also present in epidermal cells adjacent to a surgical amputation site, and in mononuclear cells in lymphoid tissue. The other 2 pangolins had mild enteritis without crypt involvement, and no immunolabeling was detected. The nucleic acid sequences of polymerase chain reaction (PCR) amplicons from these 4 pangolins were identical to a Chinese CPV-2c strain from domestic dogs. Quantitative PCR revealed a higher ratio of CPV-2 nucleic acid to internal control gene in the 2 pangolins with severe intestinal lesions and positive immunoreactivity. Herein, we present evidence of CPV-2 infections in pangolins.
Assuntos
Doenças do Cão , Infecções por Parvoviridae , Parvovirus Canino , Animais , Animais Selvagens , Cães , Contagem de Leucócitos/veterinária , Pangolins , Infecções por Parvoviridae/veterinária , FilogeniaRESUMO
BACKGROUND: The human hookworm, Necator americanus, is a parasite that infects almost half a billion people worldwide. Although treatment is available, vaccination is favorable to combat the spread of this parasite due to its wide distribution and continuous reinfection cycle in endemic communities. METHODS: We have designed a lipopeptide oral delivery system using a B-cell epitope derived from the aspartic protease Na-APR-1 from N americanus, attached to a T-helper epitope. Lipopeptides were self-assembled into nanoparticles or entrapped in liposomes that were electrostatically coated with alginate and trimethyl chitosan polymer shields. The adjuvant-free vaccine candidates were orally administered to mice and generated a humoral immune response against both peptide antigen, and the parent protein in the hookworm gut. RESULTS: The vaccine candidates were evaluated in a rodent hookworm challenge model, resulting in up to 98% and 99% decreases in mean intestinal worm and egg burdens in immunized mice, respectively. CONCLUSIONS: Lipopeptide survived the gastrointestinal conditions, induced humoral immune responses and drived protection against parasite challenge infection.
Assuntos
Infecções por Uncinaria/prevenção & controle , Lipopeptídeos/imunologia , Vacinas/imunologia , Animais , Infecções por Uncinaria/parasitologia , Imunidade Humoral , Lipopeptídeos/metabolismo , Masculino , Camundongos , Camundongos Endogâmicos BALB C , Necator americanus/metabolismo , VacinaçãoRESUMO
Hookworms are some of the most widespread of the soil-transmitted helminths (STH) with an estimated 438.9 million people infected. Until relatively recently Ancylostoma ceylanicum was regarded as a rare cause of hookworm infection in humans, with little public health relevance. However, recent advances in molecular diagnostics have revealed a much higher prevalence of this zoonotic hookworm than previously thought, particularly in Asia. This study examined the prevalence of STH and A. ceylanicum in the municipalities of Palapag and Laoang in the Philippines utilizing real-time polymerase chain reaction (PCR) on stool samples previously collected as part of a cross-sectional survey of schistosomiasis japonica. Prevalence of hookworm in humans was high with 52.8% (n = 228/432) individuals positive for any hookworm, 34.5% (n = 149/432) infected with Necator americanus, and 29.6% (n = 128/432) with Ancylostoma spp; of these, 34 were PCR-positive for A. ceylanicum. Considering dogs, 12 (n = 33) were PCR-positive for A. ceylanicum. This is the first study to utilize molecular diagnostics to identify A. ceylanicum in the Philippines with both humans and dogs infected. Control and elimination of this zoonotic hookworm will require a multifaceted approach including chemotherapy of humans, identification of animal reservoirs, improvements in health infrastructure, and health education to help prevent infection.
Assuntos
Ancylostoma/isolamento & purificação , Ancilostomíase/epidemiologia , Ancilostomíase/veterinária , Doenças do Cão/epidemiologia , Adolescente , Adulto , Idoso , Ancilostomíase/parasitologia , Animais , Criança , Pré-Escolar , Doenças do Cão/parasitologia , Cães , Fezes/parasitologia , Feminino , Humanos , Masculino , Pessoa de Meia-Idade , Filipinas/epidemiologia , Prevalência , Reação em Cadeia da Polimerase em Tempo Real/veterinária , Solo/parasitologia , Adulto JovemRESUMO
Human hookworm infections caused by adult Ancylostoma spp. and Necator americanus are one of the most important tropical diseases. We performed a survey of intestinal helminths using the Kato-Katz fecal examination technique targeting 1,156 villagers residing in 2 northern provinces (Preah Vihear and Stung Treng) of Cambodia in 2018. The results revealed a high overall egg positive rate of intestinal helminths (61.9%), and the egg positive rate of hookworms was 11.6%. Nine of the hookworm egg positive cases in Preah Vihear Province were treated with 5-10 mg/kg pyrantel pamoate followed by purging with magnesium salts, and a total of 65 adult hookworms were expelled in diarrheic stools. The adult hookworms were analyzed morphologically and molecularly to confirm the species. The morphologies of the buccal cavity and dorsal rays on the costa were observed with a light microscope, and the nucleotide sequences of mitochondrial cytochrome c oxidase subunit 1 (cox1) gene were analyzed. The majority of the hookworm adults (90.7%) were N. americanus, whereas the remaining 9.3% were Ancylostoma ceylanicum, a rare hookworm species infecting humans. The results revealed a high prevalence of hookworm infections among people in a northern part of Cambodia, suggesting the necessity of a sustained survey combined with control measures against hookworm infections.
Assuntos
Ancylostoma/isolamento & purificação , Infecções por Uncinaria/diagnóstico , Infecções por Uncinaria/parasitologia , Enteropatias Parasitárias/diagnóstico , Enteropatias Parasitárias/parasitologia , Necator americanus/isolamento & purificação , Patologia Molecular/métodos , População Rural , Adulto , Ancylostoma/genética , Ancylostoma/ultraestrutura , Animais , Camboja/epidemiologia , Complexo IV da Cadeia de Transporte de Elétrons , Fezes/parasitologia , Feminino , Infecções por Uncinaria/epidemiologia , Infecções por Uncinaria/prevenção & controle , Humanos , Enteropatias Parasitárias/epidemiologia , Masculino , Microscopia , Necator americanus/genética , Necator americanus/ultraestrutura , Prevalência , Adulto JovemRESUMO
Caenorhabditis elegans is a free-living nematode that resides in soil and typically feeds on bacteria. We postulate that haematophagic C. elegans could provide a model to evaluate vaccine responses to intestinal proteins from hematophagous nematode parasites, such as Necator americanus. Human erythrocytes, fluorescently labelled with tetramethylrhodamine succinimidyl ester, demonstrated a stable bright emission and facilitated visualization of feeding events with fluorescent microscopy. C. elegans were observed feeding on erythrocytes and were shown to rupture red blood cells upon capture to release and ingest their contents. In addition, C. elegans survived equally on a diet of erythrocytes. There was no statistically significant difference in survival when compared with a diet of Escherichia coli OP50. The enzymes responsible for the digestion and detoxification of haem and haemoglobin, which are key components of the hookworm vaccine, were found in the C. elegans intestine. These findings support our postulate that free-living nematodes could provide a model for the assessment of neutralizing antibodies to current and future hematophagous parasite vaccine candidates.
Assuntos
Caenorhabditis elegans/fisiologia , Vacinas/imunologia , Animais , Sangue , Dieta , Modelos Animais de Doenças , Comportamento Alimentar , Microscopia de FluorescênciaRESUMO
BACKGROUND: Abdominal obesity and presence of the metabolic syndrome (MetS) significantly increase the risk of developing diseases such as Type 2 diabetes mellitus (T2DM) with escalating emergence of MetS and T2DM constituting a significant public health crisis worldwide. Lower prevalence of inflammatory and metabolic diseases such as T2DM in countries with higher incidences of helminth infections suggested a potential role for these parasites in the prevention and management of certain diseases. Recent studies confirmed the potential protective nature of helminth infection against MetS and T2DM via immunomodulation or, potentially, alteration of the intestinal microbiota. This Phase 1b safety and tolerability trial aims to assess the effect of inoculation with helminths on physical and metabolic parameters, immune responses, and the microbiome in otherwise healthy women and men. METHODS: Participants eligible for inclusion are adults aged 18-50 with central obesity and a minimum of one additional feature of MetS recruited from the local community with a recruitment target of 54. In a randomised, double-blind, placebo-controlled design, three groups will receive either 20 or 40 stage three larvae of the human hookworm Necator americanus or a placebo. Eligible participants will provide blood and faecal samples at their baseline and 6-monthly assessment visits for a total of 24 months with an optional extension to 36 months. During each scheduled visit, participants will also undergo a full physical examination and complete diet (PREDIMED), physical activity, and patient health (PHQ-9) questionnaires. Outcome measurements include tolerability and safety of infection with Necator americanus, changes in metabolic and immunological parameters, and changes in the composition of the faecal microbiome. DISCUSSION: Rising cost of healthcare associated with obesity-induced metabolic diseases urgently calls for new approaches in disease prevention. Findings from this trial will provide valuable information regarding the potential mechanisms by which hookworms, potentially via alterations in the microbiota, may positively influence metabolic health. TRIAL REGISTRATION: The protocol was registered on ANZCTR.org.au on 05 June 2017 with identifier ACTRN12617000818336. Alternatively, a Google search using the above trial registration number will yield a direct link to the trial protocol within the ANZCTR website.
Assuntos
Síndrome Metabólica/terapia , Necatoríase , Obesidade/complicações , Terapia com Helmintos/métodos , Adolescente , Adulto , Animais , Diabetes Mellitus Tipo 2/prevenção & controle , Método Duplo-Cego , Fezes/microbiologia , Feminino , Microbioma Gastrointestinal/fisiologia , Humanos , Imunomodulação , Larva , Masculino , Síndrome Metabólica/imunologia , Síndrome Metabólica/microbiologia , Pessoa de Meia-Idade , Necator americanus , Necatoríase/imunologia , Necatoríase/microbiologia , Placebos , Queensland , Terapia com Helmintos/efeitos adversos , Resultado do TratamentoRESUMO
Background & objectives: Different coproculture techniques have been developed for culturing the hookworm (HW) larvae for morphological identification in the resource-limited settings. The objective of this study was to compare the performances of Harada-Mori culture (HMC), agar plate culture (APC) and modified APC (MAPC) of HW positive stool specimens for identification of HW species in East Sikkim. Methods: This prospective study was done in East Sikkim from May 2015 to May 2016. Stool and blood samples were collected from paediatric patients with gastrointestinal symptoms. The HW positive stool specimens by microscopy were subjected to HMC, APC and MAPC techniques to harvest HW larvae. Stoll's dilution egg count for determining egg intensity and blood parameters were performed in all the 12 HW-positive patients. Results: Twelve of the 180 samples were found positive for HW by microscopy and predominance of Necator americanus (75%) over Ancylostoma duodenale (25%) was observed. Blood parameters results showed high pack cell volume (PCV) values in 78.6 per cent, anaemia in 75 per cent and high eosinophil count in most patients. Stoll's dilution egg count showed moderate infection in 66.6 per cent, light and heavy infections in 16.7 per cent each. Interpretation & conclusions: Our results showed that APC yielded 100 per cent results and was easier to perform in the laboratory compared to MAPC and HMC techniques.
RESUMO
During 2012-2015, US-bound refugees living in Myanmar-Thailand border camps (n = 1,839) were surveyed for hookworm infection and treatment response by using quantitative PCR. Samples were collected at 3 time points: after each of 2 treatments with albendazole and after resettlement in the United States. Baseline prevalence of Necator americanus hookworm was 25.4%, Ancylostoma duodenale 0%, and Ancylostoma ceylanicum (a zoonosis) 5.4%. Compared with N. americanus prevalence, A. ceylanicum hookworm prevalence peaked in younger age groups, and blood eosinophil concentrations during A. ceylanicum infection were higher than those for N. americanus infection. Female sex was associated with a lower risk for either hookworm infection. Cure rates after 1 dose of albendazole were greater for A. ceylanicum (93.3%) than N. americanus (65.9%) hookworm (p<0.001). Lower N. americanus hookworm cure rates were unrelated to ß-tubulin single-nucleotide polymorphisms at codons 200 or 167. A. ceylanicum hookworm infection might be more common in humans than previously recognized.
Assuntos
Ancylostoma/isolamento & purificação , Ancilostomíase/epidemiologia , Ancilostomíase/parasitologia , Refugiados , Adolescente , Adulto , Idoso , Idoso de 80 Anos ou mais , Albendazol/uso terapêutico , Ancilostomíase/tratamento farmacológico , Animais , Anti-Helmínticos/uso terapêutico , Criança , Pré-Escolar , Fezes/parasitologia , Feminino , Humanos , Lactente , Masculino , Pessoa de Meia-Idade , Mianmar/epidemiologia , Tailândia/epidemiologia , Adulto JovemRESUMO
Hookworm is a major public health concern, yet still relatively little is known about the immunological responses involved in human infection. Animal studies are mainly confined to using the natural rodent helminth Nippostrongylus brasiliensis as this has been proposed as the most accurate model of hookworm infection in the mouse, with both its life cycle and the immune responses it invokes having been extremely well characterized. In this review, we examine the roles that type 2 innate lymphoid cells (ILC2s) play in immunity and host tolerance to hookworm infection, particularly N. brasiliensis. This includes their role in the initiation and regulation of immune responses, as well as in the resolution and limitation of tissue damage required after an infection with a large organism, such as a helminth.
Assuntos
Ancylostomatoidea/imunologia , Citocinas/imunologia , Infecções por Uncinaria/imunologia , Imunidade Inata/imunologia , Nippostrongylus/imunologia , Células Th2/imunologia , Animais , Modelos Animais de Doenças , Feminino , Infecções por Uncinaria/parasitologia , Humanos , Masculino , Camundongos , Doenças Negligenciadas/imunologia , Doenças Negligenciadas/parasitologiaRESUMO
Despite the profound health implications of Necator americanus infection in humans, many aspects of its interaction with the host immune system are poorly understood. Here we investigated the early events at the interface of N. americanus larvae (L3) and human dendritic cells (DCs). Our data show that co-culturing DCs and the larvae trigger ex-sheathing of hookworms rapidly where a majority of DCs are sequestered onto the larval sheath allowing the ex-sheathed larvae to migrate away unchallenged. Intriguingly, DCs show negligible interaction with the ex-sheathed larvae, alluding to differences between the surface chemistry of the larva and its sheath. Furthermore, blocking of two key C-type lectin receptors on DC surface (i.e. DC-SIGN and mannose receptor) resulted in inhibition of ex-sheathing process and DC sequestration, highlighting the importance of C-type lectins on DCs in the induction of the ex-sheathing. Analyses of DC phenotype and cytokine profile after co-culture with the N. americanus larvae showed an immature phenotype as evidenced by the low expression of the maturation markers and cytokines. These data provide new insights into early events at the interface of human DCs and N. americanus larvae and could explain how L3 evade immune recognition upon initial interaction with DCs.
Assuntos
Células Dendríticas/imunologia , Interações Hospedeiro-Parasita/imunologia , Evasão da Resposta Imune , Larva/fisiologia , Necator americanus/fisiologia , Animais , Antígenos de Helmintos/imunologia , Moléculas de Adesão Celular/antagonistas & inibidores , Células Cultivadas , Citocinas/imunologia , Células Dendríticas/parasitologia , Humanos , Larva/imunologia , Lectinas Tipo C/antagonistas & inibidores , Receptor de Manose , Lectinas de Ligação a Manose/antagonistas & inibidores , Necator americanus/imunologia , Necatoríase/imunologia , Receptores de Superfície Celular/antagonistas & inibidoresRESUMO
Hookworm infections are widely prevalent in tropical and subtropical areas, especially in low income regions. In the body, hookworms parasitize the proximal small intestine, leading to chronic intestinal hemorrhage and iron deficiency anemia. Occasionally, hookworms can cause overt gastrointestinal bleeding, but this is often ignored in heavily burdened individuals from endemic infectious areas. A total of 424 patients with overt obscure gastrointestinal bleeding were diagnosed by numerous blood tests or stool examinations as well as esophagogastroduodenoscopy, colonoscopy, capsule endoscopy or double-balloon enteroscopy. All of the patients lived in hookworm endemic areas and were not screened for hookworm infection using sensitive tests before the final diagnosis. The patients recovered after albendazole treatment, blood transfusion, and iron replacement, and none of the patients experienced recurrent bleeding in the follow-up. All the 31 patients were diagnosed with hookworm infections without other concomitant bleeding lesions, a rate of 7.3% (31/424). Seventeen out of 227 patients were diagnosed with hookworm infections in the capsule endoscopy (CE), and 14 out of 197 patients were diagnosed with hookworm infections in the double balloon enteroscopy (DBE). Hookworm infections can cause overt gastrointestinal bleeding and should be screened in patients with overt obscure gastrointestinal bleeding (OGIB) in endemic infectious areas with sensitive methods. Specifically, the examination of stool specimens is clinically warranted for most patients, and the proper examination for stool eggs relies on staff's communication.
Assuntos
Hemorragia Gastrointestinal/etiologia , Hemorragia Gastrointestinal/parasitologia , Infecções por Uncinaria/complicações , Infecções por Uncinaria/parasitologia , Adulto , Idoso , Albendazol/uso terapêutico , Ancylostomatoidea/isolamento & purificação , Anemia Ferropriva/diagnóstico , Anemia Ferropriva/etiologia , Anemia Ferropriva/parasitologia , Anemia Ferropriva/terapia , Animais , Anti-Helmínticos/uso terapêutico , Endoscopia por Cápsula , Endoscopia Gastrointestinal , Fezes/parasitologia , Feminino , Hemorragia Gastrointestinal/diagnóstico , Hemorragia Gastrointestinal/terapia , Infecções por Uncinaria/diagnóstico , Infecções por Uncinaria/terapia , Humanos , Masculino , Pessoa de Meia-Idade , Contagem de Ovos de Parasitas , Resultado do TratamentoRESUMO
Hookworm infections are rare causes of acute gastrointestinal bleeding. We report a middle aged man with primary nephrotic syndrome and pulmonary embolism. During the treatment with steroids and anticoagulants, the patient presented acute massive hemorrhage of the gastrointestinal tract. The results of gastroscopy showed red worms in the duodenum. Colonoscopy and CT angiogram of abdomen were unremarkable. Capsule endoscopy revealed fresh blood and multiple hookworms in the jejunum and ileum. Hookworms caused the acute intestinal bleeding. The patient responded well to albendazole. Hematochezia was markedly ameliorated after eliminating the parasites. Hence, hookworm infection should be considered in the differential diagnosis of a patient with obscure gastrointestinal bleeding. Capsule endoscopy may offer a better means of diagnosis for intestinal hookworm infections.
Assuntos
Endoscopia por Cápsula , Endoscopia Gastrointestinal , Hemorragia Gastrointestinal/diagnóstico , Hemorragia Gastrointestinal/etiologia , Infecções por Uncinaria/complicações , Doença Aguda , Albendazol/uso terapêutico , Ancylostomatoidea/isolamento & purificação , Animais , Anti-Helmínticos/uso terapêutico , Diagnóstico Diferencial , Duodeno/parasitologia , Infecções por Uncinaria/tratamento farmacológico , Infecções por Uncinaria/parasitologia , Humanos , Íleo/parasitologia , Jejuno/parasitologia , Masculino , Pessoa de Meia-Idade , Resultado do TratamentoRESUMO
Fatty acid and retinol-binding proteins (FARs) comprise a family of unusual α-helix rich lipid-binding proteins found exclusively in nematodes. They are secreted into host tissues by parasites of plants, animals and humans. The structure of a FAR protein from the free-living nematode Caenorhabditis elegans is available, but this protein [C. elegans FAR-7 (Ce-FAR-7)] is from a subfamily of FARs that does not appear to be important at the host/parasite interface. We have therefore examined [Necator americanus FAR-1 (Na-FAR-1)] from the blood-feeding intestinal parasite of humans, N. americanus. The 3D structure of Na-FAR-1 in its ligand-free and ligand-bound forms, determined by NMR (nuclear magnetic resonance) spectroscopy and X-ray crystallography respectively, reveals an α-helical fold similar to Ce-FAR-7, but Na-FAR-1 possesses a larger and more complex internal ligand-binding cavity and an additional C-terminal α-helix. Titration of apo-Na-FAR-1 with oleic acid, analysed by NMR chemical shift perturbation, reveals that at least four distinct protein-ligand complexes can be formed. Na-FAR-1 and possibly other FARs may have a wider repertoire for hydrophobic ligand binding, as confirmed in the present study by our finding that a range of neutral and polar lipids co-purify with the bacterially expressed recombinant protein. Finally, we show by immunohistochemistry that Na-FAR-1 is present in adult worms with a tissue distribution indicative of possible roles in nutrient acquisition by the parasite and in reproduction in the male.
Assuntos
Interações Hospedeiro-Parasita , Necator americanus/metabolismo , Necatoríase/metabolismo , Proteínas de Ligação ao Retinol/metabolismo , Animais , Sítios de Ligação , Caenorhabditis elegans/metabolismo , Caenorhabditis elegans/patogenicidade , Proteínas de Caenorhabditis elegans/química , Proteínas de Caenorhabditis elegans/metabolismo , Proteínas de Transporte/química , Proteínas de Transporte/metabolismo , Ácidos Graxos/química , Ácidos Graxos/metabolismo , Ligantes , Necator americanus/química , Necator americanus/patogenicidade , Necatoríase/parasitologia , Reprodução , Proteínas de Ligação ao Retinol/químicaRESUMO
Hookworm infection is still prevalent in southern Thailand despite control measures. Hookworm eggs submerged for an extended period under water from rainfall or in latrines may not survive, but they may recover their ability to develop into infective larvae when exposed to atmospheric air. This study examined the survival of the hookworm eggs in stool suspension and the restoration of development capability after prolonged storage. In stool mass, eggs developed normally and yielded infective filariform larvae (FL) in 7 days. On the contrary, in 1:10 stool suspension, hookworm eggs were found to remain at the 4-8 cell stage; degenerated eggs were observed after 15 days of storage, and the number of degenerated eggs reached 80 % on day 30. Aeration of the suspension, or transferring to a Petri dish or agar plate, restored the capacity of eggs stored for up to 15 days to develop into FL; thereafter, the capacity declined sharply. Retardation of egg development under water or in stool suspension may be due to a lack of atmospheric air. Use of "night soil" from latrines as fertilizer may be one factor in maintaining hookworm transmission, as worm eggs can undergo normal development upon exposure to atmospheric air.
Assuntos
Ancylostomatoidea/crescimento & desenvolvimento , Fezes/parasitologia , Infecções por Uncinaria/parasitologia , Necator/crescimento & desenvolvimento , Necatoríase/parasitologia , Preservação Biológica/métodos , Ancylostomatoidea/patogenicidade , Animais , Feminino , Infecções por Uncinaria/epidemiologia , Infecções por Uncinaria/transmissão , Humanos , Larva , Necator/patogenicidade , Necatoríase/epidemiologia , Necatoríase/transmissão , Óvulo/crescimento & desenvolvimento , Preservação Biológica/normas , Prevalência , Solo/parasitologia , Suspensões , Tailândia/epidemiologia , Água/parasitologiaRESUMO
CONTEXT: Dichapetalum filicaule Breteler (Dichapetalaceae) is a rare species occurring only in Côte d'Ivoire and Ghana. Although research on several species of the genus has produced interesting bioactive compounds, particularly the Dichapetalins, a novel class of triterpenoids with antineoplastic properties, there is virtually no information on the ethnobotanical uses and chemical constituents of D. filicaule. OBJECTIVE: The phytochemical and anthelminthic activities of the constituents of D. filicaule were investigated. MATERIALS AND METHODS: Chemical constituents of the petroleum ether, chloroform-acetone, and methanol root extracts of D. filicaule were isolated by column chromatography and characterized by their physico-chemical properties, 1-D and 2-D NMR spectroscopy and mass spectrometry. In vitro anthelminthic activity of the extracts and compounds against the human hookworm, Necator americanus, Stiles 1902 (Nematoda: Ancylostomatidae) was determined within a concentration range of 2500-250 µg/ml using the Egg Hatch Inhibition (EHI) Assay. The hookworm species were identified using a published polymerase chain reaction (PCR) method. RESULTS: A new dichapetalin, dichapetalin X (1), together with the known dichapetalin A (2), pomolic acid (3), glycerol monostearate (4), D:A-friedooleanan-3ß-ol (5), and D:A-friedooleanan-3-one (6) were isolated. Compounds 1, 2, and 4 exhibited EHI with IC50 values of 523.2, 162.4, and 306.0 µg/ml, respectively, against the hookworm. The positive control albendazole gave an IC50 value of 93.27 µg/ml. DISCUSSION AND CONCLUSION: This is the first report of the phytochemical investigation of D. filicaule. The study has yielded a new dichapetalin and also demonstrated the potential anthelminthic properties of the constituents.
Assuntos
Anti-Helmínticos/farmacologia , Compostos Heterocíclicos de 4 ou mais Anéis/farmacologia , Magnoliopsida , Necator americanus/efeitos dos fármacos , Extratos Vegetais/farmacologia , Compostos de Espiro/farmacologia , Adolescente , Adulto , Idoso , Idoso de 80 Anos ou mais , Animais , Anti-Helmínticos/isolamento & purificação , Criança , Pré-Escolar , Cromatografia em Camada Fina , Feminino , Compostos Heterocíclicos de 4 ou mais Anéis/isolamento & purificação , Humanos , Espectroscopia de Ressonância Magnética , Magnoliopsida/química , Masculino , Espectrometria de Massas , Pessoa de Meia-Idade , Necator americanus/genética , Necator americanus/crescimento & desenvolvimento , Contagem de Ovos de Parasitas , Testes de Sensibilidade Parasitária , Fitoterapia , Extratos Vegetais/isolamento & purificação , Raízes de Plantas , Plantas Medicinais , Solventes/química , Compostos de Espiro/isolamento & purificação , Adulto JovemRESUMO
Na-ASP-2 is an efficacious hookworm vaccine antigen. However, despite elucidation of its crystal structure and studies addressing its immunobiology, the function of Na-ASP-2 has remained elusive. We probed a 9000-protein human proteome microarray with Na-ASP-2 and showed binding to CD79A, a component of the B-cell antigen receptor complex. Na-ASP-2 bound to human B lymphocytes ex vivo and downregulated the transcription of approximately 1000 B-cell messenger RNAs (mRNAs), while only approximately 100 mRNAs were upregulated, compared with control-treated cells. The expression of a range of molecules was affected by Na-ASP-2, including factors involved in leukocyte transendothelial migration pathways and the B-cell signaling receptor pathway. Of note was the downregulated transcription of lyn and pi3k, molecules that are known to interact with CD79A and control B-cell receptor signaling processes. Together, these results highlight a previously unknown interaction between a hookworm-secreted protein and B cells, which has implications for helminth-driven immunomodulation and vaccine development. Further, the novel use of human protein microarrays to identify host-pathogen interactions, coupled with ex vivo binding studies and subsequent analyses of global gene expression in human host cells, demonstrates a new pipeline by which to explore the molecular basis of infectious diseases.
Assuntos
Ancylostomatoidea/imunologia , Linfócitos B/imunologia , Infecções por Uncinaria/imunologia , Receptores de Células Precursoras de Linfócitos B/imunologia , Proteoma/imunologia , Proteínas Recombinantes/imunologia , Transdução de Sinais/imunologia , Adulto , Animais , Antígenos de Helmintos/imunologia , Antígenos CD79/imunologia , Células Cultivadas , Regulação para Baixo/genética , Regulação para Baixo/imunologia , Proteínas de Helminto/imunologia , Interações Hospedeiro-Patógeno/genética , Interações Hospedeiro-Patógeno/imunologia , Humanos , Leucócitos Mononucleares/imunologia , Pessoa de Meia-Idade , Fosfatidilinositol 3-Quinases/genética , Fosfatidilinositol 3-Quinases/imunologia , Análise Serial de Proteínas/métodos , Proteoma/genética , RNA Mensageiro/genética , RNA Mensageiro/imunologia , Transdução de Sinais/genética , Transcrição Gênica/genética , Transcrição Gênica/imunologia , Regulação para Cima/genética , Regulação para Cima/imunologia , Quinases da Família src/genética , Quinases da Família src/imunologiaRESUMO
BACKGROUND: Estimation of prevalence of Strongyloides stercoralis infection is required in endemic areas, in order to identify areas in need of control programmes. Data on prevalence of strongyloidiasis in Madagascar are scant. Aim of this work was to estimate prevalence of S. stercoralis in four districts of Madagascar. METHODS: Fecal and serum samples collected in the context of a previous study on schistosomiasis were tested with S. stercoralis real-time PCR and serology, respectively. A multiplex real-time PCR for Ascaris lumbricoides, Ancylostoma duodenalis, Necator americanus, and Trichuris trichiura was done on fecal samples collected in the areas demonstrating higher prevalence of strongyloidiasis. Comparisons between proportions were made using Fisher exact test, with false discovery rate correction used for post-hoc comparisons. A multivariable Firth logistic regression model was used to assess potential risk factors for S. stercoralis infection. RESULTS: Overall, 1775 serum samples were tested, of which 102 of 487 (20.9%) and 104 of 296 (35.2%) were serological-positive in Marovoay and in Vatomandry districts (both coastal areas), respectively, compared to 28 of 496 (5.6%) and 30 of 496 (6.1%) in Tsiroanomandidy and in Ambositra districts (both highlands), respectively (adj. p < 0.001). PCR for S. stercoralis was positive in 15 of 210 (7.1%) and in 11 of 296 (3.7%) samples from Marovoay from Vatomandry, respectively, while was negative for all samples tested in the other two districts. High prevalence of A. lumbricoides (45.9%), hookworm (44.6%) and T. trichiura (32.1%) was found in Vatomandry. In the multivariable analysis, strongyloidiasis was associated with hookworm infection. Hookworm infection was also associated with male sex and lower education level. CONCLUSIONS: S. stercoralis prevalence proved higher in coastal areas compared to highlands. Different climatic conditions may explain this distribution, along with previous rounds of anthelminthics distributed in the country, which may have reduced the parasite load in the population. The high prevalence of the other soil-transmitted helminths (STH) in Vatomandry was unexpected, given the good coverage with benzimidazole in control campaigns. Further studies are needed to explore the risk factors for STH and S. stercoralis infections in Madagascar, in order to align with the WHO recommendations.
RESUMO
Necator americanus is the major cause of human hookworm infection, which is a global cause of anemia in the developing world. Ongoing efforts to control hookworm infection include the identification of candidate vaccine antigens as well as potential therapeutic targets from the infective L3 larval stages and adult stages of the parasite. One promising family of proteins are the adult-stage-secreted cytosolic glutathione S-transferases (GSTs). Nematode GSTs facilitate the inactivation and degradation of a variety of electrophilic substrates (drugs) via the nucleophilic addition of reduced glutathione. Parasite GSTs also play significant roles in multi-drug resistance and the modulation of host immune defense mechanisms. Here, the structure of Na-GST-3, one of three GSTs secreted by adult-stage N. americanus, is reported. Unlike most GST structures, the Na-GST-3 crystal contains a monomer in the asymmetric unit. However, the monomer forms a prototypical GST dimer across the crystallographic twofold. A glutathione from the fermentation process is bound to the monomer. The overall binding cavity of Na-GST-3 is reminiscent of that of other N. americanus GSTs and is larger and capable of binding a wider array of ligands than GSTs from organisms that have other major detoxifying mechanisms. Furthermore, despite having low sequence identity to the host GST, Na-GST-3 has a greater tertiary-structure similarity to human sigma-class GST than was observed for the other N. americanus GSTs.