RESUMO
Extracting genomic DNA of pathogenic agents from formalin-fixed specimens is inherently difficult. Storage of samples in formalin results in nucleic acid cross-linking and DNA fragmentation. In this study, DNA was extracted from 45 Giardia-positive stool samples stored in formalin and subjected to PCR amplification targeting the triose phosphate isomerase (tpi), beta gardin (bg) and glutamate dehydrogenase (gdh) genes. Samples were rehydrated by using a descending alcohol series before DNA extraction using a commercial kit. This was followed by EDTA-mediated inhibition of DNase activity and prolonged treatment with proteinase K to digest contaminating proteins. DNA was amplified at rates of 64.4% (29/45) at the tpi, 40% (18/45) at the bg and 20% (9/45) at the gdh loci as seen on nested PCR. DNA quality was subsequently tested in a genotyping experiment which produced high-quality sequences at the tpi (41.2%; 12/29) bg (50%; 9/18), and gdh (22.2%; 2/9) loci and enabled differentiation of Giardia strains at the subtype level. The modified extraction protocol was effective at removing inhibitors and reversing cross-linking of DNA. However, PCR amplification was limited to short fragments of DNA which resulted in highest success rate on amplification of the shortest (334 bp) gene fragment tested.
Assuntos
DNA de Protozoário/isolamento & purificação , Fezes/parasitologia , Fixadores/efeitos adversos , Formaldeído/efeitos adversos , Giardia/genética , Sequência de Bases , Proteínas do Citoesqueleto/genética , DNA de Protozoário/química , DNA de Protozoário/genética , DNA de Protozoário/normas , Etanol/administração & dosagem , Genótipo , Técnicas de Genotipagem , Giardia/química , Giardia/classificação , Giardia/enzimologia , Glutamato Desidrogenase/genética , Humanos , Reação em Cadeia da Polimerase , Proteínas de Protozoários/genética , Solventes/administração & dosagem , Fatores de Tempo , Triose-Fosfato Isomerase/genéticaRESUMO
Objectives: Toxocarosis involving cranial nerves is extremely rare and almost exclusively concerns the optic nerve. Toxocarosis involving the seventh cranial nerve has not been reported. Case report: A 33y male developed left-sided Bell's palsy two days after left-sided otalgia 6y before. Despite extensive diagnostic work-up at that time the cause of Bell's palsy remained unknown. During the following years Bell's palsy slightly improved but retromandibular pain remained almost unchanged and he developed enlarged lymph nodes along the jugular veins, submandibularly, and in the trigonum caroticum. Re-evaluation 6y later revealed an increased titer of serum antibodies against Toxocara canis and a positive Westernblot for Toxocara canis ES-antigen. Despite absent eosinophilia in the serum, toxocarosis was diagnosed and a therapy with albendazole initiated, with benefit for retromandibular pain, but hardly for Bell's palsy or enlarged lymph nodes. CSF investigations after albendazole revealed a positive Westernblot for antibodies against toxocara but absent pleocytosis or eosinophilia, and negative PCR for Toxocara canis. Conclusions: Visceral larva migrans due to Toxocara canis may be associated with Bell's palsy, retromandibular pain, and lymphadenopathy. A causal relation between Bell's palsy and the helminthosis remains speculative. Adequate therapy years after onset of the infestation may be of limited benefit.
Objetivos: La toxocarosis que compromete los nervios craneales es extremadamente infrecuente y afecta casi exclusivamente al nervio óptico. No ha sido comunicada la toxocarosis que afecte al séptimo nervio cranial por lo que se expone un caso clínico. Caso clínico: Varón de 33 años que había presentado seis años antes, una otalgia izquierda seguida dos días más tarde de una parálisis de Bell ipsilateral. A pesar de un exhaustivo estudio, la causa de esta parálisis quedó sin etiología. En los años siguientes la parálisis mejoró moderadamente pero persistió un dolor retromandibular casi sin variaciones y el paciente desarrolló adenopatías en la cadena yugular, submandi-bulares y en el triángulo carotídeo. Una re-evaluación efectuada seis años más tarde detectó un título elevado de anticuerpos contra Toxocara canis y un antígeno ES de T. canis positivo mediante Westernblot. A pesar de no haber eosinofilia en la sangre, se diagnosticó una toxoca-rosis iniciándose terapia con albendazol, con lo cual se observó una reducción del dolor retro-mandibular aunque escaso beneficio sobre la parálisis de Bell y los ganglios comprometidos. El líquido cefalorraquídeo analizado después de haberse efectuado la cura con albendazol reveló un test de Westernblot positivo para anticuerpos anti-toxocara, en ausencia de pleocitosis o eosinofilia, y una RPC para Toxocara canis negativa. Conclusiones: El síndrome de larva migrans visceral debido a T. canis puede asociarse con parálisis de Bell, dolor retroman-dibular y linfoadenopatías. La relación causal entre la parálisis de Bell y la helmintosis permanece en el plano especulativo. La terapia anti-parasitaria realizada años tras la instalación de una infestación puede tener un beneficio aunque limitado.
Assuntos
Adulto , Humanos , Masculino , Paralisia de Bell/parasitologia , Dor Facial/parasitologia , Toxocaríase/complicações , Paralisia de Bell/diagnóstico , Dor Facial/diagnóstico , Toxocaríase/diagnósticoRESUMO
OBJECTIVES: Toxocarosis involving cranial nerves is extremely rare and almost exclusively concerns the optic nerve. Toxocarosis involving the seventh cranial nerve has not been reported. CASE REPORT: A 33y male developed left-sided Bell's palsy two days after left-sided otalgia 6y before. Despite extensive diagnostic work-up at that time the cause of Bell's palsy remained unknown. During the following years Bell's palsy slightly improved but retromandibular pain remained almost unchanged and he developed enlarged lymph nodes along the jugular veins, submandibularly, and in the trigonum caroticum. Re-evaluation 6y later revealed an increased titer of serum antibodies against Toxocara canis and a positive Westernblot for Toxocara canis ES-antigen. Despite absent eosinophilia in the serum, toxocarosis was diagnosed and a therapy with albendazole initiated, with benefit for retromandibular pain, but hardly for Bell's palsy or enlarged lymph nodes. CSF investigations after albendazole revealed a positive Westernblot for antibodies against toxocara but absent pleocytosis or eosinophilia, and negative PCR for Toxocara canis. CONCLUSIONS: Visceral larva migrans due to Toxocara canis may be associated with Bell's palsy, retromandibular pain, and lymphadenopathy. A causal relation between Bell's palsy and the helminthosis remains speculative. Adequate therapy years after onset of the infestation may be of limited benefit.
Assuntos
Paralisia de Bell/parasitologia , Dor Facial/parasitologia , Toxocaríase/complicações , Adulto , Paralisia de Bell/diagnóstico , Dor Facial/diagnóstico , Humanos , Masculino , Toxocaríase/diagnósticoRESUMO
Capillaria hepatica (syn. for Calodium hepaticum) is a zoonotic nematode parasitizing in the livers of rodents as main hosts and in numerous other mammals including humans. It is the causative agent of the rare conditions of hepatic capillariosis and spurious C. hepatica infections in humans. In this review, 163 reported cases of infestations with this parasite (72 reports of hepatic capillariosis, 13 serologically confirmed infestations and 78 observations of spurious infections) are summarized with an overview on the distribution, symptoms, pathology, diagnosis, serology and therapy of this rare human pathogen.
Assuntos
Capillaria/fisiologia , Infecções por Enoplida , Hepatomegalia/parasitologia , Estágios do Ciclo de Vida/fisiologia , Fígado/parasitologia , Doenças dos Roedores , Adulto , África , Animais , Anti-Helmínticos/administração & dosagem , Anti-Helmínticos/uso terapêutico , Anticorpos Anti-Helmínticos/análise , Anticorpos Anti-Helmínticos/imunologia , Biópsia , Capillaria/efeitos dos fármacos , Criança , Pré-Escolar , Reservatórios de Doenças , Infecções por Enoplida/diagnóstico , Infecções por Enoplida/epidemiologia , Infecções por Enoplida/imunologia , Infecções por Enoplida/mortalidade , Infecções por Enoplida/parasitologia , Infecções por Enoplida/patologia , Infecções por Enoplida/fisiopatologia , Infecções por Enoplida/terapia , Ensaio de Imunoadsorção Enzimática , Europa (Continente) , Feminino , Hepatomegalia/imunologia , Hepatomegalia/patologia , Hepatomegalia/fisiopatologia , Humanos , Lactente , Estágios do Ciclo de Vida/efeitos dos fármacos , Fígado/imunologia , Fígado/patologia , Fígado/fisiopatologia , Masculino , América do Norte , Filogeografia , Doenças dos Roedores/diagnóstico , Doenças dos Roedores/epidemiologia , Doenças dos Roedores/imunologia , Doenças dos Roedores/mortalidade , Doenças dos Roedores/parasitologia , Doenças dos Roedores/patologia , Doenças dos Roedores/fisiopatologia , Roedores , América do Sul , Taxa de Sobrevida , Tomografia Computadorizada por Raios XRESUMO
We report a 51-year-old male with a history of palpitations, hepatopathy and hypercholesterolemia, who habitually ate raw goat meat, and developed general fasciculations, muscle cramps in the lower limbs, distal muscle weakness and wasting, without upper motor neuron signs or sensory abnormalities. Diagnostic workup revealed positive antibodies against Toxocara canis in the serum and cerebrospinal fluid. Nerve conduction studies revealed a proximal and distal axonal lesion of motor nerves and needle electromyography was indicative of acute and chronic denervation with giant motor unit action potentials. Despite a therapy with albendazole and riluzole, muscle weakness and wasting further progressed and affected also the respiratory muscles. Followup nerve conduction studies and electromyography confirmed progression of the axonal degeneration. Whether lower motor neuron disease was causally related to neurotoxocariasis or due to a general metabolic defect, remains speculative.
Assuntos
Helmintíase do Sistema Nervoso Central/complicações , Doença dos Neurônios Motores/parasitologia , Toxascaríase/complicações , Toxocara canis , Animais , Humanos , Masculino , Pessoa de Meia-Idade , Toxocara canis/imunologia , Toxocara canis/isolamento & purificaçãoRESUMO
We report a 51-year-old male with a history of palpitations, hepatopathy and hypercholesterolemia, who habitually ate raw goat meat, and developed general fasciculations, muscle cramps in the lower limbs, distal muscle weakness and wasting, without upper motor neuron signs or sensory abnormalities. Diagnostic workup revealed positive antibodies against Toxocara canis in the serum and cerebrospinal fuid. Nerve conduction studies revealed a proximal and distal axonal lesion of motor nerves and needle electromyography was indicative of acute and chronic denervation with giant motor unit action potentials. Despite a therapy with albendazole and riluzole, muscle weakness and wasting further progressed and affected also the respiratory muscles. Followup nerve conduction studies and electromyography confrmed progression of the axonal degeneration. Whether lower motor neuron disease was causally related to neurotoxocariasis or due to a general metabolic defect, remains speculative.
Presentamos un hombre de 51 años con una historia de palpitaciones, hepatopatía e hipercolesterolemia que comía habitualmente carne de cabra cruda, que desarrolló un cuadro caracterizado por fasciculaciones generalizadas, calambres musculares en las extremidades inferiores, pérdida de fuerza y atrofa muscular distal, sin signos de lesión de motoneurona superior o alteraciones sensitivas. El laboratorio mostró anticuerpos anti Toxocara canis en suero y líquido cefalorraquídeo. La electromiografía mostró una lesión axonal proximal y distal de nervios motores y denervación con potenciales de acción gigantes en las unidades motoras. A pesar de tratarse con albendazol y riluzona, la debilidad muscular y atrofa continuaron progresando. Una nueva electromiografía confrmó la progresión de la degeneración axonal. La asociación entre esta enfermedad de motoneurona inferior y la neurotoxocariasis, es especulativa.
Assuntos
Animais , Humanos , Masculino , Pessoa de Meia-Idade , Helmintíase do Sistema Nervoso Central/complicações , Doença dos Neurônios Motores/parasitologia , Toxascaríase/complicações , Toxocara canis , Toxocara canis/imunologia , Toxocara canis/isolamento & purificaçãoRESUMO
Cutaneous leishmaniasis of the New World, in particular when caused by Leishmania (L.) braziliensis, harbours the risk of lymphogenic as well as hematogenic dissemination. This may result in mucocutaneous leishmaniasis causing severe destruction of orofacial structures. Dissemination may occur years after the disappearance of the skin lesions. In contrast, cutaneous leishmaniasis of the old world, is typically restricted to the site of inoculation. Therefore, a conservative diagnostic and therapeutic approach is usually sufficient. Infections acquired in the new world should be treated systemically, if infection with Leishmania (Viannia) braziliensis complex cannot be excluded. Here we report on three Austrian soldiers, who, weeks after having participated in an international jungle patrol course in Belize, presented themselves with multiple ulcers on the upper limbs. Diagnosis of cutaneous leishmaniasis was made based upon histological evaluation of biopsies taken from several ulcers revealing the presence of leishmanial bodies, and detection of amastigote leishmania in smears of material obtained from the ulcers. As species phenotyping could not be performed, infection with L. brasiliensis as well as progression into a mucocutaneous form were possible, demanding systemic therapy. Several treatment options including local cryotherapy with liquid nitrogen, paromomycin (Humatis Pulvis, Parke-Davis) 15% topically or oral fluconazole (Diflucan, Pfizer) 200 mg/d were applied, but showed no effect. Hence, a systemic therapy with intravenous pentamidine (Pentacarinat, Gerot), three times in total, 3-4 mg/kg body weight each, led to a complete regression of the lesions within four weeks.
Assuntos
Leishmaniose Cutânea/diagnóstico , Leishmaniose Cutânea/terapia , Militares , Viagem , Adulto , Áustria , Belize , Humanos , MasculinoRESUMO
Infection of humans with embryonated eggs of Toxocara canis (larva migrans) remains asymptomatic, or results in covert or common toxocarosis, visceral larva migrans syndrome, or ophthalmologic and neurologic impairment. Though neurological manifestations of Toxocara canis larvae are rare, toxocarosis remains an important differential diagnosis of various neurological disorders. Manifestations of the central nervous system are dementia, meningo-encephalitis, myelitis, cerebral vasculitis, epilepsy, or optic neuritis. Manifestations of the peripheral nervous system comprise radiculitis, affection of cranial nerves, or musculo-skeletal involvement. If toxocarosis is neglected, ignored, or refused as a differential of these abnormalities, it may be easily overlooked for years. Early recognition and treatment of the infection is, however, of paramount importance since it reduces morbidity and mortality and the risk of secondary superinfection. Like the visceral manifestations, neurological manifestations of toxocarosis are treated by benzimidazole components, most frequently albendazole, corticosteroids, or diethylcarbamazine. If detected and treated early, the prognosis of neurological manifestations of toxocarosis is favourable.
Assuntos
Helmintíase do Sistema Nervoso Central/parasitologia , Doenças do Sistema Nervoso Periférico/parasitologia , Toxocara canis , Toxocaríase/diagnóstico , Animais , Gatos , Helmintíase do Sistema Nervoso Central/diagnóstico , Helmintíase do Sistema Nervoso Central/tratamento farmacológico , Diagnóstico Diferencial , Cães , Humanos , Doenças do Sistema Nervoso Periférico/diagnóstico , Doenças do Sistema Nervoso Periférico/tratamento farmacológico , Toxocaríase/complicações , Toxocaríase/tratamento farmacológicoRESUMO
Infection of humans with embryonated eggs of Toxocara canis (larva migrans) remains asymptomatic, or results in covert or common toxocarosis, visceral larva migrans syndrome, or ophthalmologic and neurologic impairment. Though neurological manifestations of Toxocara canis larvae are rare, toxocarosis remains an important differential diagnosis of various neurological disorders. Manifestations of the central nervous system are dementia, meningo-encephalitis, myelitis, cerebral vasculitis, epilepsy, or optic neuritis. Manifestations of the peripheral nervous system comprise radiculitis, affection of cranial nerves, or musculo-skeletal involvement. If toxocarosis is neglected, ignored, or refused as a differential of these abnormalities, it may be easily overlooked for years. Early recognition and treatment of the infection is, however, of paramount importance since it reduces morbidity and mortality and the risk of secondary superinfection. Like the visceral manifestations, neurological manifestations of toxocarosis are treated by benzimidazole components, most frequently albendazole, corticosteroids, or diethylcarbamazine. If detected and treated early, the prognosis of neurological manifestations of toxocarosis is favourable.
Infecção humana com ovos embrionados de Toxocara canis (larva migrans) pode permanecer assintomática ou resultar em toxocaríase acentuada ou comum, síndrome da larva migrans visceral ou manifestações neurológicas ou oftalmológicas. Embora manifestações neurológicas das larvas de Toxocara canis sejam raras, a toxocaríase permanece como importante diagnóstico diferencial de várias manifestações neurológicas. Manifestações do sistema nervoso central são demência, meningoencefalite, mielite, vasculite cerebral, epilepsia, ou neurite ótica. Manifestações do sistema nervoso periférico compreendem radiculite, agressão de nervos cranianos ou envolvimento músculo-esquelético. Se a toxocaríase é negligenciada, ignorada, ou recusada como diferencial destas anormalidades, ela pode ser facilmente desapercebida por anos. Reconhecimento precoce de tratamento da infecção é portanto de fundamental importância uma vez que reduz sua morbidade e mortalidade e o risco de superinfecção secundária. Da mesma maneira que as manifestações viscerais, as neurológicas são tratadas por benzimidazólicos, mais freqüentemente albendazole, corticosteróides ou dietilcarbamazine. Se detectado e tratado precocemente, o prognóstico das manifestações neurológicas da toxocaríase é favorável.