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1.
Acta Trop ; 254: 107195, 2024 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-38548212

RESUMO

A clinical case of an adult horse with invasive, ulcerative, proliferative, pyogranulomatous disease of the skin (tumor) in the shoulder region is presented. The mass had a granulomatous and crater-shaped appearance, with serosanguinous discharge and the presence of fistulas with caseous material. The tumor was removed by surgery and sent to the laboratory for diagnosis. Histopathology was performed using Grocott-Gomori methenamine silver stain. The presence of necrotic material, fibrosis, infiltrated cells, and brown-colored hyphae, characteristic of members of the genus Pythium, were observed. To identify the infecting species, conventional PCRs for the amplification of the ITS-1 was carried out. Histopathological and PCR tests confirmed infection by a Pythium insidiosum strain closely associated with previous records from the US and Central America. Our report represents the first molecularly confirmed case of equine pythiosis in Mexico.


Assuntos
Doenças dos Cavalos , Pitiose , Pythium , Animais , Pitiose/diagnóstico , Pitiose/microbiologia , Pitiose/patologia , Cavalos , Pythium/isolamento & purificação , Pythium/genética , Pythium/classificação , Doenças dos Cavalos/parasitologia , Doenças dos Cavalos/microbiologia , Doenças dos Cavalos/diagnóstico , México , Reação em Cadeia da Polimerase , DNA Espaçador Ribossômico/genética , Masculino , Histocitoquímica , Pele/patologia , Pele/microbiologia , Pele/parasitologia
2.
J Equine Vet Sci ; 132: 104976, 2024 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-38056727

RESUMO

Horse pythiosis is considered an endemic disease in the Brazilian Pantanal region, causing devastating health and economic losses. This study aimed to enhance the understanding of pythiosis epidemiology, map the distribution of horse body lesions, and investigate the correlation between these lesions and warm body surface areas, potentially implicating hematophagous vectors in the disease's transmission. A prospective study was conducted on equids in the Pantanal Mato-grossense and adjacent areas from 2012 to 2022, with 112 horses and three mules diagnosed with pythiosis. Clinical and epidemiological data, lesions' photographic records, and healthy equids' thermal imaging were collected. Most pythiosis cases occurred between January and March, correlating with regional flood cycles. Most lesions were found on limbs and the ventral abdomen, with dark-colored horses exhibiting a higher frequency of lesions. Interestingly, the thermal mapping revealed that warm areas on a healthy horse's body overlapped significantly with lesion distribution - blood-sucking insects also prefer these areas. The results suggest that pythiosis lesions in horses correlate with warmer areas of the animal body, reinforcing the hypothesis of vector involvement in disease transmission. This study underscores the need for further observational research to fully understand the complex epidemiological dynamics of pythiosis in horses.


Assuntos
Doenças dos Cavalos , Parasitos , Pitiose , Cavalos , Animais , Doenças dos Cavalos/epidemiologia , Doenças dos Cavalos/patologia , Pitiose/epidemiologia , Pitiose/patologia , Brasil/epidemiologia , Estudos Prospectivos
3.
J Comp Pathol ; 195: 34-50, 2022 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-35817539

RESUMO

Pythiosis is an endemic disease in northeastern Brazil and we now report the epidemiological, clinical and pathological findings in a retrospective study of naturally occurring cases in domestic animals. From January 1985 to December 2020, the Laboratory of Animal Pathology of the Federal University of Campina Grande examined 13,542 tissue samples from necropsies and biopsies. Among these samples, 306 were diagnosed as pythiosis: 195 cases in horses, 75 in sheep, 19 in dogs, six in mules, four in cattle, three in cats, two in goats, one in a donkey and one in an ostrich. Affected equids had lesions in the skin, mammary glands and nasal cavities. Affected sheep had cutaneous, nasal and digestive lesions while cattle and goats had cutaneous lesions. Carnivores developed lesions, mainly in the alimentary tract, of sufficient severity to cause death or result in euthanasia. The single affected bird had lesions in the alimentary tract and surgical excision resulted in remission. The disease had a long and life-threatening clinical course in most affected species but resolved spontaneously in cattle. Clinical signs were directly related to the location of the lesions, which were invariably characterized by chronic inflammation associated with intralesional hyphae. Veterinary clinicians and pathologists should be familiar with the clinicopathological features of pythiosis and the wide range of susceptible animal species.


Assuntos
Doenças do Gato , Doenças dos Bovinos , Doenças do Cão , Pitiose , Pythium , Doenças dos Ovinos , Animais , Animais Domésticos , Brasil/epidemiologia , Gatos , Bovinos , Doenças dos Bovinos/epidemiologia , Cães , Equidae , Pitiose/epidemiologia , Pitiose/patologia , Estudos Retrospectivos , Ovinos
4.
Pesqui. vet. bras ; 40(8): 647-650, Aug. 2020. ilus
Artigo em Inglês | LILACS, VETINDEX | ID: biblio-1135673

RESUMO

Pythiosis is an emerging infectious disease affecting captive and free-ranging wild animals. We report granulomatous pneumonia due to Pythium insidiosum in two South American coatis (Nasua nasua), who were found dead without any clinical records. Severe granulomatous pneumonia associated with pleural effusion was revealed in the necropsy. Microscopically, variably sized granulomas and pyogranulomas presented negative hyphae profiles at the periphery of their necrotic cores. Grocott methenamine silver stain highlighted these structures, and immunostain (anti- P. insidiosum) was strongly positive. Molecular analysis by polymerase chain reaction amplified P. insidiosum specific DNA. These findings characterized P. insidiosum as a cause of granulomatous pneumonia in coatis and proved that pythiosis needs to be considered in the differential diagnosis of respiratory diseases affecting this species in endemic areas.(AU)


A pitiose é uma doença infecciosa emergente que afeta animais silvestres de cativeiro e em vida livre. Reportamos dois casos de pneumonia granulomatosa decorrentes da infecção por Pythium Insidiosum em quatis sul-americanos (Nasua nasua), que foram encontrados mortos sem apresentar nenhum quadro clínico prévio. Pneumonia granulomatosa severa associada a efusão pleural foi observada durante a necropsia. Na microscopia, foram observados múltiplos granulomas e piogranulomas de tamanhos variados que continham imagens negativas de hifas na periferia de seus centros necróticos. A coloração de metenamina de prata (Grocott) evidenciou estas estruturas, e a imunomarcação (anti-P. insidiosum) foi fortemente positiva. A análise molecular pela reação de polimerase em cadeia amplificou o DNA específico do P. insidiousum. Estes achados caracterizaram o P. insidiosum como a causa da pneumonia granulomatosa nos quatis e provou que a pitiose deve ser considerada um diagnostico diferencial para outras doenças respiratórias que afetam esta espécie.(AU)


Assuntos
Animais , Masculino , Feminino , Pneumonia/etiologia , Pneumonia/patologia , Pneumonia/veterinária , Procyonidae , Pitiose/complicações , Pitiose/patologia
5.
Pesqui. vet. bras ; 40(5): 340-345, May 2020. ilus
Artigo em Inglês | VETINDEX, LILACS | ID: biblio-1135633

RESUMO

The epidemiological, clinical and pathological aspects of cutaneous pythiosis occurring in cattle from three farms in the Northeastern of Brazil are described. A biopsy of the lesions of one bovine from each farm was performed. In two cases, the affected cattle had contact with water accumulated in dams during the dry season in the semiarid region. Another case occurred in the coastal tropical region in cattle grazing around irrigation channels. Clinically, lesions were observed mainly on the skin of the thoracic and/or pelvic limbs, characterized by flat and irregular ulcerated areas or nodules of varying sizes, some with fistulous tracts penetrating deep into the subcutaneous tissue. In one case the regional lymph nodes were affected. Histologically, in all cases, pyogranulomatous dermatitis associated with negative hyphae images, in hematoxylin-eosin stained sections, were observed. In sections stained by Grocott methenamine silver, the hyphae measured 2-8μm and had irregular ramifications and rare septations. Immunohistochemistry technique demonstrated strong immunolabeling for Pythium insidiosum. Pythiosis should be included in the differential diagnosis of dermatopathies in cattle in the Northeastern of Brazil.(AU)


Descrevem-se os aspectos epidemiológicos, clínicos e patológicos da pitiose cutânea em bovinos de três propriedades do Nordeste do Brasil. Uma biópsia das lesões de um bovino de cada propriedade foi realizada. Em dois casos, os bovinos afetados tiveram acesso à água acumulada em açudes durante a estação seca da região semiárida. O outro bovino acometido estava a pastoreio próximo a canais de irrigação na região litorânea. Clinicamente, as lesões foram observadas principalmente na pele dos membros torácicos e/ou pélvicos e caracterizavam-se por áreas planas e irregulares de ulceração ou nódulos de tamanhos variados, alguns com trajetos fistulosos penetrando profundamente no tecido subcutâneo. Em um caso, os linfonodos regionais foram afetados. Histologicamente, em todos os casos, observou-se dermatite piogranulomatosa associada a imagens negativas de hifas, em secções corados por hematoxilina e eosina. Em seções coradas por metenamina de prata de Grocott, as hifas mediam 2-8μm e possuíam ramificações irregulares com raras septações. A imuno-histoquímica demonstrou forte imunomarcação para Pythium insidiosum. A pitiose deve ser incluída como diagnóstico diferencial de dermatopatias de bovinos no Nordeste do Brasil.(AU)


Assuntos
Animais , Bovinos , Doenças dos Bovinos/epidemiologia , Dermatite/veterinária , Pitiose/diagnóstico , Pitiose/patologia , Pitiose/epidemiologia , Pythium
6.
J Mycol Med ; 30(1): 100919, 2020 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-31901425

RESUMO

We evaluated the in vitro activity of miltefosine against 29 Pythium spp. and the in vivo therapeutic response of 2mg/kg/day of miltefosine given orally to rabbit with pythiosis induced experimentally. The MICs (in µg/mL) of miltefosine was medium-dependent and ranged from 0.5 to 2 and 32-64 on RPMI 1640 and Mueller Hinton broth, respectively. The treatment with miltefosine demonstrated significantly lower subcutaneous lesion areas compared to the control group but was not sufficient for the complete remission of the lesions. This study indicates that miltefosine has limited efficacy against pythiosis and furthers in vitro and in vivo studies are necessary to determine the possible potential of this drug in the treatment of pythiosis.


Assuntos
Antifúngicos/uso terapêutico , Dermatomicoses/tratamento farmacológico , Fosforilcolina/análogos & derivados , Pitiose/tratamento farmacológico , Animais , Dermatomicoses/microbiologia , Dermatomicoses/patologia , Modelos Animais de Doenças , Progressão da Doença , Relação Dose-Resposta a Droga , Feminino , Humanos , Testes de Sensibilidade Microbiana , Fosforilcolina/uso terapêutico , Pitiose/microbiologia , Pitiose/patologia , Pythium/isolamento & purificação , Pythium/patogenicidade , Coelhos , Tela Subcutânea/microbiologia , Resultado do Tratamento
7.
Curr Eye Res ; 45(5): 542-549, 2020 05.
Artigo em Inglês | MEDLINE | ID: mdl-31593643

RESUMO

Purpose: To describe the clinical characteristics and histopathological features in a rabbit model of Pythium insidiosum keratitis.Methods: Zoospores of P. insidiosum isolated from a patient with microbial keratitis were used for inoculation of the right eye of 48 New Zealand White rabbits in either low (LD) or high dose (HD). Apart from variable dosage the rabbits were grouped (6 rabbits per group) based on route of inoculation (topical on abraded cornea or intracorneal) and immunosuppression (subconjunctival steroid or no steroid). Left eye received phosphate buffered saline via route similar to the right eye. Daily clinical examination of the eye was done, the corneas were harvested on day 3, 7 and 9 and part of the cornea was preserved in 10% neutral buffered formalin for histopathological examination.Results: Left eye of all rabbits were clinically normal. Eyes with intracorneal injection of zoospores developed infection irrespective of dose of inoculation and administration of steroids. One of the consistent early signs of infection was ring like infiltrate in the peripheral cornea. On day 2, rabbits receiving HD developed significantly greater inflammation compared to LD [median clinical score in HD- 11 (IQR = 10-12), versus 9 (IQR = 8-9) in LD (p = 0.004)]. The density of inflammation showed temporal correlation (increase with time) when the inoculum was low. Of the rabbits that received topical inoculation one rabbit cornea showed mild infiltrate in steroid group while no eye was infected in the group without steroid. Sparsely septate to aseptate branching filaments were noted in the stroma of all infected corneas.Conclusions: We describe the first animal model of Pythium keratitis that holds promise for future studies. While topical inoculation of zoospores was unsuccessful in causing infection intracorneal inoculation without immunosuppression was sufficient to develop clinically severe keratitis in rabbits.


Assuntos
Úlcera da Córnea/patologia , Modelos Animais de Doenças , Infecções Oculares Parasitárias/patologia , Pitiose/patologia , Pythium/isolamento & purificação , Animais , Córnea/parasitologia , Úlcera da Córnea/diagnóstico , Infecções Oculares Parasitárias/diagnóstico , Pitiose/diagnóstico , Coelhos , Microscopia com Lâmpada de Fenda , Corpo Vítreo/parasitologia
8.
Mycopathologia ; 185(5): 801-812, 2020 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-31845178

RESUMO

Pythiosis is an emerging infectious disease caused by the aquatic oomycete Pythium insidiosum, a fungal-like organism. It is believed that P. insidiosum's zoospores, its infected form, play major role in pathogenesis. Vascular and ocular infections are the most common clinical manifestation in humans. It is difficult to establish the diagnosis given its relatively rarity and difficulty to distinguish P. insidiosum from other molds. Delay in diagnosis and treatment has been associated with poor outcomes. High index of suspicion is the key, particularly in thalassemia patients with arterial insufficiency and patients with fungal keratitis/endophthalmitis without improvement on antifungal therapy. Tissue culture and zoospore induction remain gold standard for diagnosis; however, DNA-based method should be performed simultaneously. The combination of radical surgery, antifungal agents, and immunotherapy has been recommended. It was previously believed that surgery with negative surgical margins was the essential to survive in vascular pythiosis; however, it was recently found that patients could have residual disease despite documented negative surgical margins as infected clot may be dislodged to proximal arterial sites prior to surgery. Serum ß-D-glucan (BG) has been used to monitor disease response after treatment initiation in vascular pythiosis. A significant decrease in BG levels within 2 weeks after surgery is indicative of the absence of residual infection. Unfortunately, monitoring tools for ocular pythiosis are not yet available. Itraconazole plus terbinafine have generally been used in P. insidiosum-infected patients; however, antibacterial agents, including azithromycin and linezolid, have also been used with favorable outcomes in ocular disease. Recently, azithromycin or clarithromycin plus doxycyclin were used in two relapsed vascular pythiosis patients with good outcomes.


Assuntos
Pitiose , Pythium , Antibacterianos/uso terapêutico , Antifúngicos/farmacologia , Doenças Transmissíveis Emergentes/diagnóstico , Doenças Transmissíveis Emergentes/terapia , Doenças Transmissíveis Emergentes/transmissão , Combinação de Medicamentos , Infecções Oculares Fúngicas/diagnóstico , Infecções Oculares Fúngicas/terapia , Imunoterapia/métodos , Itraconazol/farmacologia , Oomicetos , Patologia Molecular , Pitiose/diagnóstico , Pitiose/patologia , Pitiose/terapia , Pitiose/transmissão , Pythium/efeitos dos fármacos , Pythium/isolamento & purificação , Testes Sorológicos , Esporos Fúngicos/isolamento & purificação , Terbinafina/farmacologia , Talassemia/complicações , Lesões do Sistema Vascular/diagnóstico , Lesões do Sistema Vascular/microbiologia , Lesões do Sistema Vascular/terapia , beta-Glucanas/sangue
9.
Vet Clin Pathol ; 48(1): 83-88, 2019 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-30897244

RESUMO

A 2-year-old female spayed Boxer dog was presented for a 1-month history of progressive hemorrhagic diarrhea with tenesmus and weight loss despite trial courses of antibiotics and diet change. Abdominal ultrasound revealed severe, focal thickening, and loss of normal architecture of the colonic wall with abdominal lymphadenomegaly. Dry-mount fecal cytology, performed on several consecutive days, consistently revealed numerous, round, 16-20 µm structures with basophilic, granular content, and a thin cell wall. Transmission electron microscopy identified these structures as fungi. Culture, polymerase chain reaction (PCR), and sequencing of the internal transcribed spacer, D1/D2 regions, and DNA-directed RNA polymerase II core subunit (RPB2) confirmed the presence of Basidiobolus microsporus in the feces. Biopsies collected via ileocolonoscopy revealed marked, multifocal, chronic, neutrophilic, and eosinophilic ileitis and colitis with ulceration, granulation tissue, and intralesional hyphae (identified with Gomori methenamine silver stain). A Pythium enzyme-linked immunosorbent assay and Pythium-specific PCR performed on the formalin-fixed paraffin-embedded biopsy specimens were positive while Basidiobolus-specific PCR was negative, thus confirming a diagnosis of pythiosis. This report describes a fatal case of colonic and intestinal pythiosis with the presence of fecal Basidiobolus sp. spores, suggestive of concurrent gastrointestinal basidiobolomycosis.


Assuntos
Coinfecção/veterinária , Doenças do Cão/microbiologia , Entomophthorales , Gastroenteropatias/veterinária , Pitiose/diagnóstico , Pythium , Zigomicose/veterinária , Animais , Coinfecção/diagnóstico , Coinfecção/microbiologia , Coinfecção/patologia , Doenças do Cão/diagnóstico , Doenças do Cão/patologia , Cães , Feminino , Gastroenteropatias/diagnóstico , Gastroenteropatias/microbiologia , Gastroenteropatias/patologia , Pitiose/complicações , Pitiose/microbiologia , Pitiose/patologia , Zigomicose/complicações , Zigomicose/diagnóstico , Zigomicose/patologia
10.
J Zoo Wildl Med ; 49(3): 784-787, 2018 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-30212327

RESUMO

Pythium insidiosum, an aquatic oomycete, causes chronic lesions in the skin and digestive tract of multiple species. A captive-bred Bactrian camel ( Camelus bactrianus) showed clinical signs of lethargy and weight loss in a clinical course of 30 days, with no response to treatment. At necropsy, the abdominal cavity had approximately 32 L of a yellow, turbid fluid with fibrin. The third compartment of the stomach (C-3) showed a focal area of rupture covered with fibrin. Close to this area, the C-3 wall was thickened and firm, demonstrating irregular, yellow, and friable areas on cut surface (kunkers). Microscopically, these corresponded to necrosis, characterized by a central amorphous eosinophilic material, surrounded by a pyogranulomatous inflammatory infiltrate and fibrosis. Negatively stained hyphae were observed at the periphery of the necrotic areas, which showed marked immunostaining for P. insidiosum. Pythiosis in camelids may involve the stomach, resulting in peritonitis and death.


Assuntos
Camelus , Pitiose/patologia , Gastropatias/veterinária , Animais , Evolução Fatal , Masculino , Gastropatias/microbiologia , Gastropatias/patologia
11.
Br J Ophthalmol ; 102(12): 1629-1633, 2018 12.
Artigo em Inglês | MEDLINE | ID: mdl-29545414

RESUMO

BACKGROUND: Pythium insidiosum is a parafungus that causes keratitis resembling fungal keratitis. This study compares outcome in a large cohort of patients with P insidiosum keratitis treated with antifungal drugs, to a pilot group treated with antibacterial antibiotics. METHODS: Between January 2014 and December 2016, 114 patients with culture positive P insidiosum keratitis were included in the study. A subset of culture isolates was tested in vitro for response to nine antibacterial antibiotics by disc diffusion and E test. Patients were treated with topical natamycin in 2014, 2015 and up until mid 2016. Thereafter, the patients received a combination of topical linezolid and topical and oral azithromycin. Therapeutic penetrating keratoplasty (TPK) was done for patients not responding to medical therapy. RESULTS: In vitro disc diffusion assay showed linezolid to be most effective. The rate of TPK was significantly higher in 2015 compared with 2016 (43/45, 95.6% vs 22/32, 68.8%; p=0.002). Eighteen patients were treated with antibacterial and 14 were treated with antifungal antibiotic in 2016. One patient was lost to follow-up in each group. The rate of TPK was higher and proportion of healed ulcers was lower (p=0.21, Fisher's exact test) in the group on antifungal therapy (TPK-11/13, 84.6%; Healed-2/13, 15.3%) compared with the group on antibacterial therapy (TPK-11/17, 64.7%; Healed-6/17, 35.2%). CONCLUSIONS: We report favourable but not statistically significant response of P insidiosum keratitis to antibacterial agents in a pilot series of patients. Further evaluation of this strategy in larger number of patients is recommended.


Assuntos
Antibacterianos/uso terapêutico , Antifúngicos/uso terapêutico , Surtos de Doenças/prevenção & controle , Infecções Oculares Fúngicas/tratamento farmacológico , Ceratite/tratamento farmacológico , Pitiose/tratamento farmacológico , Pythium/efeitos dos fármacos , Adulto , Infecções Oculares Fúngicas/patologia , Feminino , Humanos , Ceratite/patologia , Ceratoplastia Penetrante/estatística & dados numéricos , Masculino , Pessoa de Meia-Idade , Projetos Piloto , Pitiose/patologia , Pythium/patogenicidade , Resultado do Tratamento
12.
Mycoses ; 61(2): 104-110, 2018 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-28972292

RESUMO

Pythiosis is a severe disease caused by Pythium insidiosum. Currently, the research on the treatment of pythiosis uses rabbits as an experimental infection model. To reduce the use of animals in scientific experimentation, alternative models are increasingly necessary options. The objective of this study was to establish a new experimental infection model for pythiosis using embryonated chicken eggs. First, we tested the inoculation of 4 zoospore concentrations into the egg allantoic cavity at 3 embryonic days. We observed that increased zoospore concentration causes a decrease in survival time, and at a later embryonic day (the 14th) of infection, embryos showed delayed mortality. To confirm the reproducibility of the model, we chose the 14th embryonic day for the inoculation of 50 zoospores/egg, and the experiment was repeated twice. Mortality began with 30% embryos 48 hours after inoculation, and 95% embryos died within 72 hours. There was no mortality in the uninfected control group. The infection was confirmed by culture, PCR and histopathology. Immunohistochemistry confirmed the presence of hyphae in blood vessels in the umbilical cords in 95% of embryos and only 1 liver (5%). Our results suggest that embryonated eggs can be a very useful alternative infection model to study pythiosis.


Assuntos
Modelos Animais de Doenças , Pitiose/patologia , Pythium/crescimento & desenvolvimento , Pythium/patogenicidade , Animais , Embrião de Galinha , Histocitoquímica , Imuno-Histoquímica , Técnicas Microbiológicas , Reação em Cadeia da Polimerase , Reprodutibilidade dos Testes , Análise de Sobrevida , Fatores de Tempo
13.
Cornea ; 36(9): 1124-1132, 2017 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-28582375

RESUMO

PURPOSE: To elucidate the histopathology of Pythium insidiosum keratitis and to describe a novel, simple, and rapid staining technique for identification of oomycete Pythium insidiosum and to differentiate it from fungi. METHODS: This is a laboratory investigation study of 38 nonconsecutive cases (37 ocular samples and 1 colonic biopsy); 14 microbiologically diagnosed as Pythium insidiosum keratitis and 24 as fungal keratitis. Review of clinical, demographic details, microbiological results, and identification of cases that necessitated evisceration was performed. Reevaluation of histopathology slides was done using stains such as hematoxylin-eosin, Gomori methenamine silver (GMS), periodic acid-Schiff (PAS), potassium iodide-sulfuric acid (IKI-H2SO4). Morphology, degree, and nature of inflammation and load, distribution, and staining results of Pythium insidiosum and its comparison with fungi were studied. RESULTS: Delay in zoospore formation, failure of growth, and delay in identification of Pythium were the main cause of evisceration. Corneal pythiosis showed epithelial ulceration, stromal destruction, and varying inflammation; load and distribution of Pythium were inversely proportional to inflammation. The filaments were commonly wide, with admixed narrower structures and uncommonly involved Descemet membrane. The oomycete was not discretely discerned with PAS stain and stained distinctly with GMS stain and IKI-H2SO4 stain (100% sensitive). In comparison, fungal organisms stained well with PAS and GMS stain, but not with IKI-H2SO4 stain (100% specific). CONCLUSIONS: Pythium insidiosum keratitis is perhaps not more devastating than fungal keratitis but late diagnosis, misdiagnosis, and treatment as fungal infection are major heralds. Early diagnosis may markedly improve the patient outcome. IKI-H2SO4 is a cost-effective, simple, sensitive, and specific stain for the diagnosis of oomycete Pythium.


Assuntos
Técnicas de Diagnóstico Oftalmológico , Infecções Oculares Parasitárias , Ceratite , Pitiose , Coloração e Rotulagem/métodos , Adulto , Idoso , Córnea/microbiologia , Córnea/parasitologia , Infecções Oculares Parasitárias/diagnóstico , Infecções Oculares Parasitárias/patologia , Feminino , Humanos , Compostos de Iodo , Ceratite/diagnóstico , Ceratite/parasitologia , Ceratite/patologia , Masculino , Pessoa de Meia-Idade , Pitiose/diagnóstico , Pitiose/patologia , Ácidos Sulfúricos
14.
PLoS One ; 12(5): e0177868, 2017.
Artigo em Inglês | MEDLINE | ID: mdl-28542438

RESUMO

Pythiosis is a severe and life-threatening disease that affects humans and various animal species. We report a model of vascular/disseminated pythiosis occurring after subcutaneous inoculation of 2 x 104 Pythium insidiosum zoospores/mL in immunocompromised BALB/c mice. For this model, we carried out two rounds of experiments. First, we evaluated two protocols of immunosuppression before inoculation: cyclophosphamide at 150 mg/kg (CYP group) and cyclophosphamide 200 mg/kg plus hydrocortisone acetate at 250 mg/kg (CYP+HCA group). It was not possible to obtain mortality in the CYP group; however, the combination of CYP+HCA altered disease outcomes, with mortality rates reaching 60%. Second, we used the CYP+HCA immunosuppression protocol to analyze the histological and immunological statuses triggered by disease. When we inoculated immunocompetent mice with P. insidiosum zoospores, self-healing occurred via increased levels of IL-2, IFN-γ and IL-17A, which are characteristic of the Th1/Th17 cytokine response. For infected and immunosuppressed mice, the cytokine profiles showed high levels of IL-10, IL-6 and TNF-α. Increased IL-10 values are related to fungal infection susceptibility and led us to speculate that infection may be established through suppression of the host immune response. In addition, histopathological evaluation of the kidneys and liver demonstrated the presence of hyphae and the cellular findings suggested an acute vascular inflammation that mimics vascular/disseminated pythiosis in humans. This is the first murine model for pythiosis that is useful both for understanding the pathogenesis of this disease and for evaluating new treatment approaches.


Assuntos
Ciclofosfamida/toxicidade , Hidrocortisona/análogos & derivados , Modelos Teóricos , Pitiose/etiologia , Pitiose/patologia , Pythium/imunologia , Animais , Citocinas/metabolismo , Combinação de Medicamentos , Feminino , Humanos , Hidrocortisona/toxicidade , Camundongos , Camundongos Endogâmicos BALB C , Pitiose/metabolismo , Pythium/efeitos dos fármacos
15.
Ann Clin Microbiol Antimicrob ; 16(1): 7, 2017 Feb 23.
Artigo em Inglês | MEDLINE | ID: mdl-28231795

RESUMO

BACKGROUND: Pythium insidiosum is the etiological agent of pythiosis, an emerging life-threatening infectious disease in tropical and subtropical regions. The pathogen is a fungus-like organism resistant to antifungal therapy, for this reason, most cases need extensive surgical debridments as treatment, but depending on the size and anatomical region of the lesion, such approach is unfeasible. We investigate the fungicidal effect and toxicity of crude bark extract of Stryphnodendron adstringens and commercially available tannin on Pythium insidiosum both in vitro and in vivo. METHODS: Standardized fragments of mycelia of fifteen isolates of P. insidiosum were tested with different concentrations of bark extract (10 to 30% v/v) and tannin (0.5, 1.0 and 1.5 mg/mL). For in vivo study, fifteen rabbits were experimentally infected with zoospores of P. insidiosum and treated by oral and intralesional applications of bark extract and tannin. Acute toxicity tests with both substances were also performed in rats. RESULTS: In vitro studies showed fungicidal effect for both substances at different concentrations and the SEM showed alteration on the cell wall surface of the pathogen. All infected rabbits developed a firm nodular mass that reached around 90 mm2 ninety days after inoculation, but neither the intralesional inoculation of tannin, nor the oral administration of crude extract and tannin were able to promote remission of the lesions. CONCLUSIONS: Lesions developed by rabbits presented an encapsulated abscess being quite different of naturally acquired pythiosis, which is characterized by ulcerated lesions. Since no toxicity was observed in rats or rabbits inoculated with these products, while in vitro experiments showed direct antifungal effect, therapeutic activity of S. adstringens and tannin should be clinically tested as an alternative for healing wounds in naturally acquired pythiosis.


Assuntos
Antifúngicos/farmacologia , Fabaceae/química , Micélio/efeitos dos fármacos , Pitiose/tratamento farmacológico , Pythium/efeitos dos fármacos , Taninos/farmacologia , Administração Oral , Animais , Modelos Animais de Doenças , Relação Dose-Resposta a Droga , Humanos , Injeções Intralesionais , Masculino , Micélio/crescimento & desenvolvimento , Micélio/ultraestrutura , Casca de Planta/química , Extratos Vegetais/química , Pitiose/microbiologia , Pitiose/patologia , Pythium/crescimento & desenvolvimento , Pythium/ultraestrutura , Coelhos , Ratos , Ratos Wistar
16.
J Comp Pathol ; 155(2-3): 126-129, 2016.
Artigo em Inglês | MEDLINE | ID: mdl-27406311

RESUMO

Two cases of nasal pythiosis are reported in horses from the semi-arid region of northeastern Brazil. From January 1986 to December 2015, the Laboratory of Animal Pathology, Federal University of Campina Grande received 830 equine samples, 156 (18.79%) of which were diagnosed with pythiosis. Of these, two horses (1.28%), a male and a female adult cross-breed, had lesions in the nasal cavity. Both horses had access to water reservoirs. Clinically, they had swelling in the rhinofacial region and a serosanguineous nasal discharge. Macroscopically, in case 1, the lesion affected the nasal vestibule, extending to the alar cartilage and nasal septum. In case 2, the lesion extended through the turbinates and the meatuses of the nasal cavity, as well as the ethmoid region. In both cases, the lesions were characterized by having a yellow-grey granular surface with cavitations of different sizes containing coral-like masses of necrotic tissue (kunkers). Histologically, multifocal necrotizing eosinophilic rhinitis associated with hyphae (2-8 µm) similar to Pythium insidiosum were observed. In case 2, the lesions extended to the muscle, cartilage and bone adjacent to the nasal cavity and lungs. The diagnosis was confirmed by immunohistochemistry. It is concluded that nasal pythiosis occurs sporadically in horses in the semi-arid region of northeastern Brazil where cutaneous pythiosis is prevalent.


Assuntos
Doenças dos Cavalos/microbiologia , Doenças dos Cavalos/patologia , Doenças Nasais/microbiologia , Doenças Nasais/patologia , Pitiose/patologia , Animais , Feminino , Cavalos , Masculino , Cavidade Nasal/patologia
17.
Antimicrob Agents Chemother ; 60(1): 87-91, 2016 01.
Artigo em Inglês | MEDLINE | ID: mdl-26459895

RESUMO

The present study investigated the in vitro and the in vivo interactions among azithromycin, clarithromycin, minocycline, and tigecycline against Pythium insidiosum. In vitro antimicrobial activities were determined by the broth microdilution method in accordance with CLSI document M38-A2, and the antibiotic interactions were assayed using the checkerboard MIC format. In vivo efficacy was determined using a rabbit infection model. The geometric mean MICs of azithromycin, clarithromycin, minocycline, and tigecycline against P. insidiosum were, respectively, 1.91, 1.38, 0.91, and 0.79 µg/ml. By checkerboard testing, all combinations resulted in in vitro synergistic interactions (>60%). Antagonism was not observed. The in vivo studies showed that azithromycin (20 mg/kg/day twice daily) alone or in combination with minocycline (10 mg/kg/day twice daily) significantly decreased the fungal burden. This study demonstrates that azithromycin possesses potent curative efficacy against subcutaneous pythiosis in the rabbit model.


Assuntos
Antifúngicos/farmacologia , Azitromicina/farmacologia , Claritromicina/farmacologia , Minociclina/análogos & derivados , Minociclina/farmacologia , Pitiose/tratamento farmacológico , Pythium/efeitos dos fármacos , Animais , Esquema de Medicação , Sinergismo Farmacológico , Quimioterapia Combinada , Testes de Sensibilidade Microbiana , Análise Multivariada , Pitiose/microbiologia , Pitiose/patologia , Pythium/genética , Pythium/crescimento & desenvolvimento , Pythium/isolamento & purificação , Coelhos , Pele/efeitos dos fármacos , Pele/microbiologia , Pele/patologia , Tigeciclina
18.
J Pediatric Infect Dis Soc ; 4(1): e10-3, 2015 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-26407370

RESUMO

Pythium insidiosum, also known as "swamp cancer," has been long known for its destructive effects on plants and animals. Infections with this fungus-like organism typically occur in temperate, tropical, and subtropical regions [ 1]. Human infection with P insidiosum, although exceedingly rare, is characterized by invasion of the cornea, cutaneous, subcutaneous, and orbital tissues with eosinophilic, tumor-like masses demonstrating arterial tropism [ 1, 2]. In most cases, patients have a history of recent exposure to wet environments [ 1]. If left untreated, P insidiosum is fatal, because it is an angioinvasive organism that leads to thrombosis and tissue ischemia. Reports of most human cases are from Thailand, with only 4 well documented orbital cases to date in the United States and Australia [ 1, 3, 4].


Assuntos
Doenças Orbitárias/diagnóstico , Doenças Orbitárias/terapia , Pitiose/diagnóstico , Pitiose/terapia , Pythium , Vacinas/administração & dosagem , Criança , Diagnóstico Diferencial , Olho/patologia , Face/patologia , Feminino , Humanos , Necrose/diagnóstico , Necrose/patologia , Necrose/terapia , Doenças Orbitárias/patologia , Pitiose/patologia
19.
Pesqui. vet. bras ; 35(7): 627-636, jul. 2015. tab, graf
Artigo em Português | LILACS | ID: lil-766209

RESUMO

As enfermidades que acometem a cavidade nasal de pequenos ruminantes podem causar prejuízos aos rebanhos de ovinos e caprinos na região central do Brasil. Foi realizado estudo retrospectivo dos laudos de necropsia do Laboratório de Patologia Veterinária da Universidade de Brasília (LPV-UnB) nos anos de 2003 a 2014 para verificar a ocorrência das doenças que acometeram a cavidade nasal de pequenos ruminantes. Foram analisados 463 protocolos de ovinos e 75 de caprinos totalizando 538 casos. Seis ovinos (6/463 1,29%) foram necropsiados com rinite granulomatosa micótica ou oomicótica e 22 animais do estudo (22/538; 4,08%) tiveram o diagnóstico de oestrose, sendo 86,36% ovinos e 13,64% caprinos. As rinites piogranulomatosas em ovinos ocorreram em áreas alagadas, com abundante material vegetal em decomposição. Os ovinos com pitiose rinofacial apresentaram como principais alterações aumento de volume na região nasal devido a extensas lesões granulomatosas associadas a necrose tecidual, caracterizadas por inúmeros macrófagos e polimorfonucleares circundando centros necróticos contendo o agente envolto por reação de Splendore-Hoeppli. Os ovinos com conidiobolomicose exibiram extensas áreas de necrose e inflamação piogranulomatosa, associadas à presença de hifas fúngicas na nasofaringe e também na região peribulbar e exoftalmia. A maioria dos animais com oestrose não apresentou alterações clínico-patológicas significativas, apesar de serem encontradas larvas principalmente nos seios e conchas nasais, traqueia e seio paranasal. A importância dessas enfermidades ainda é pouco conhecida na região, sendo de grande relevância que as condições clínico-patológicas e epidemiológicas sejam elucidadas para o diagnóstico, o controle e a prevenção, para evitar a expansão e prejuízos para os rebanhos...


Nasal cavity diseases that affect small ruminants can cause losses to sheep and goat herds in Central Brazil. A retrospective study of the University of Brasilia´s Veterinary Pathology Laboratory autopsy reports from 2003 to 2014 was conducted to verify the occurrence of small ruminants nasal cavity diseases. Six necropsied sheep (6/463 1.29%) showed mycotic or oomicotic granulomatous rhinitis and 22 animals (22/538, 4.08%) presented oestrosis diagnosis, affecting 86.36% of sheep and 13.64% of goats. The pyogranulomatous rhinitis in sheep occurred in flooded areas with abundant plant material decomposing. Rhinofacial pythiosis infection in animals showed as major changes swelling in the nasal region due to extensive granulomatous lesions associated with tissue necrosis, characterized by numerous macrophages and polymorphonuclear cells surrounding necrotic centers containing the agent surrounded by Splendore-Hoeppli reaction. Sheep with conidiobolomycosis showed extensive areas of necrosis and pyogranulomatous inflammation associated with fungal hyphae, localized in the nasopharynx and also in peribulbar region and exophthalmia. Most animals with oestrosis showed no significant clinical and pathological changes, even with the presence of larvae mainly in the sinuses and nasal turbinates, trachea and paranasal sinus. The importance of such diseases is still unknown in the region, and the knowledge of the clinical-pathological and epidemiological conditions is of great relevance for the diagnosis, control and prevention to avoid the expansion and losses to livestock...


Assuntos
Animais , Cavidade Nasal/patologia , Rinite/fisiopatologia , Rinite/veterinária , Ovinos , Conidiobolus/patogenicidade , Imuno-Histoquímica/veterinária , Pitiose/patologia , Pythium/patogenicidade , Reação em Cadeia da Polimerase/veterinária
20.
Pesqui. vet. bras ; 35(6): 513-517, June 2015. graf
Artigo em Inglês | LILACS | ID: lil-766190

RESUMO

This paper reports pythiosis in a sheep from southwestern Paraná, Brazil, confirmed by indirect ELISA (Enzime-Linked Immunosorbent Assay) and immunohistochemistry, as well as it describes the macro and microscopic injuries, in order to understand the pathogenicity. A 4-year-old ewe from a flock of 30 Santa Inês sheep, raised semi-extensively with access to a weir, showed cachexia, bilateral enlargement in nasal region, a serous and bloody secretion with a fetid odor from its nose and swollen submandibular and retropharyngeal lymph nodes. Blood collection was performed trough jugular vein puncture in order to make complete blood cell count (CBC) and to obtain serum for the subsequent serological examination...


Este trabalho tem como objetivo descrever um caso de pitiose confirmado por ELISA (Enzime-Linked Immunosorbent Assay) indireto e imuno-histoquímica em uma ovelha do sudoeste do Paraná, Brasil, bem como suas lesões macro e microscópicas, a fim de compreender a sua patogenicidade. Trata-se de um ovino, fêmea, de 4 anos de idade, de um rebanho de 30 animais, da raça Santa Inês, criados em sistema semiextensivo, com acesso à um açude. O animal vinha apresentando aumento de volume na região do focinho, associado a emagrecimento progressivo. No exame físico apresentou-se caquético, com aumento de volume bilateral na região nasal, e com uma secreção serosanguinolenta de odor fétido fluindo das narinas, além de possuir os linfonodos submandibulares e retrofaríngeos bilateralmente infartados. Foi realizada coleta de sangue por punção da veia jugular para realização do hemograma e obtenção do soro para posterior realização de exame sorológico....


Assuntos
Animais , Pitiose/diagnóstico , Pitiose/patologia , Ovinos , Autopsia/veterinária , Contagem de Células Sanguíneas/veterinária , Ensaio de Imunoadsorção Enzimática/veterinária , Necrose/veterinária , Pythium , Palato Duro/lesões , Perfuração do Septo Nasal/veterinária , Poluição da Água/prevenção & controle
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