Promising use of nanotechnology in Pythium insidiosum: a systematic review / Utilização promissora de nanotecnologia em Pythium insidiosum: uma revisão sistemática

ABSTRACT: The aquatic oomycete Pythium insidiosum is an emerging pathogen highly relevant in human and veterinary medicine and an etiologic agent of pythiosis, a disease of worldwide distribution mainly affecting horses, dogs, and humans, presenting cutaneous, subcutaneous, ocular, gastrointestinal, and systemic forms. The available therapeutic methods to treat this disease and its forms are not entirely effective, thus highlighting the need to investigate the forms of treatments with better efficacy, such as compounds from different pharmacological classes, compounds of natural origin, and new technological alternatives, including nanotechnology. Therefore, this study evaluated scientific publications regarding the use of nanotechnology in P. insidiosum treatment. For this, a systematic literature review, was carried out on articles published from 2010 to 2022 on the LILACS, MEDLINE, Google Scholar, PubMed, and SciELO databases using the descriptors 'Pythium insidiosum,' 'pythiosis,' 'nanotechnology,' 'nanoparticles,' 'nanoemulsion,' and 'treatment.' We reported 162 articles for the researched theme; although, only four studies were included because they met the criteria established herein. A meta-analysis was used for the statistical analysis of the data obtained in vitro studies, and we reported the use of nanotechnology can be a promising alternative in developing antimicrobial compounds with anti-P. insidiosum activity. Nevertheless, additional research is needed to verify the potential use of this technology in clinical therapy against P. insidiosum infections.

RESUMO: O oomiceto aquático Pythium insidiosum é um patógeno emergente de relevância em medicina humana e veterinária. É o agente etiológico da pitiose, uma enfermidade de distribuição mundial, que acomete principalmente em equinos, caninos e seres humanos, podendo apresentar-se nas formas cutâneas, subcutâneas, oculares, gastrointestinais e sistêmicas. Considerando que os métodos terapêuticos disponíveis para o tratamento da doença não são completamente efetivos, há uma necessidade de investigar formas de tratamentos com melhor eficácia, como os compostos de diferentes classes farmacológicas, compostos de origem natural, bem como, novas alternativas tecnológicas, incluindo a nanotecnologia. Deste modo, este trabalho objetivou avaliar publicações científicas referentes a utilização de nanotecnologia em P. insidiosum. Para isso, realizou-se uma revisão sistemática da literatura, buscando artigos no período de 2010 a 2022, nas bases de dados LILACS, MEDLINE, Google Scholar, PubMed e SciELO, utilizando-se os descritores Pythium insidiosum, pitiose, nanotecnologia, nanopartículas, nanoemulsão e tratamento. Encontrou-se 162 artigos com familiaridade a temática pesquisada; no entanto, apenas quatro estudos foram incluídos, pois atendiam os critérios estabelecidos na pesquisa. Para análise estatística dos dados obtidos nos estudos in vitro, utilizou-se meta-análise. Demonstrou-se o promissor uso de nanotecnologia como alternativa no desenvolvimento de compostos antimicrobianos com atividade anti-P. insidiosum. Entretanto, constata-se que estudos adicionais se fazem necessários para verificar o potencial uso desta tecnologia na terapêutica clínica contra infecções por P. insidiosum.

Meningoencephalitis secondary to rhinitis caused by Pythium insidiosum in sheep / Meningoencefalite secundária a rinite por Pythium insidiosum em um ovino

ABSTRACT: A case of meningoencephalitis caused by Pythium insidiosum secondary to rhinitis is reported in a three-year-old crossbred sheep from a herd of 15 animals, raised extensively and with free access to a weir. The animal presented mild dyspnea, blindness, mydriasis, opisthotonos, nystagmus, incoordination, decreased mandibular tone, and spasticity of the pelvic limbs. Macroscopic examination of the nasal cavity showed a blackish-red, irregular, friable mass that bilaterally compromised the nasal septum and the rostral portion of the nasal turbinates. In the brain, there was diffuse thickening of the leptomeninges of the cerebellum and ventral portion of the brainstem characterized by yellowish, granular material associated with vessel hyperemia. On the floor of the fourth ventricle, there was deposition of yellowish, irregular, slightly granular material that protruded towards the obex and displaced the cerebellum dorsolaterally. Microscopically, there were pyogranulomatous, eosinophilic, necrotizing rhinitis and fibrinosuppurative, eosinophilic, necrotizing meningoencephalitis, both associated with thrombosis, vasculitis, and intralesional hyphae. The hyphae were impregnated with silver and presented thin, parallel walls, rarely septate and branched. At immunohistochemistry, the hyphae were immunostained with polyclonal anti-P. insidiosum antibody in fragments of the cerebellum and nasal cavity. The findings showed that P. insidiosum rhinitis can secondarily affect the nervous system of sheep, causing nonspecific neurological clinical signs.

RESUMO: Relata-se um caso de meningoencefalite por Pythium insidiosum secundária a rinite em uma ovelha mestiça, três anos de idade, proveniente de um rebanho de 15 animais, criados extensivamente e com acesso livre a açude. O animal apresentava dispneia leve, cegueira, midríase, opistótono, nistagmo, incoordenação, diminuição do tônus mandibular e espasticidade dos membros pélvicos. O exame macroscópico da cavidade nasal evidenciou uma massa vermelho-escura, irregular, friável, que comprometia bilateralmente o septo nasal e a porção rostral dos cornetos nasais. No encéfalo, havia espessamento difuso das leptomeninges do cerebelo e porção ventral do tronco encefálico caracterizado por material granular amarelado associado à hiperemia dos vasos. No assoalho do quarto ventrículo, havia deposição de material amarelado, irregular, levemente granuloso, que se projetava em direção ao óbex e deslocava o cerebelo dorsolateralmente. Microscopicamente, havia rinite piogranulomatosa, eosinofílica, necrosante e meningoencefalite fibrinossupurativa, eosinofílica, necrosante, ambas associadas à trombose, vasculite e hifas intralesionais. Pela utilização da técnica de GMS as hifas foram impregnadas pela prata e apresentavam paredes finas e paralelas, raramente septadas e ramificadas. Na imuno-histoquímica, houve imunomarcação com o anticorpo policlonal anti-P. insidiosum em fragmentos do cerebelo e cavidade nasal. Os achados evidenciaram que rinite por P. insidiosum pode afetar secundariamente o sistema nervoso de ovinos, causando sinais clínicos neurológicos inespecíficos.

Meningoencephalitis secondary to rhinitis caused by Pythium insidiosum in sheep / Meningoencefalite secundária a rinite por Pythium insidiosum em um ovino

A case of meningoencephalitis caused by Pythium insidiosum secondary to rhinitis is reported in a three-year-old crossbred sheep from a herd of 15 animals, raised extensively and with free access to a weir. The animal presented mild dyspnea, blindness, mydriasis, opisthotonos, nystagmus, incoordination, decreased mandibular tone, and spasticity of the pelvic limbs. Macroscopic examination of the nasal cavity showed a blackish-red, irregular, friable mass that bilaterally compromised the nasal septum and the rostral portion of the nasal turbinates. In the brain, there was diffuse thickening of the leptomeninges of the cerebellum and ventral portion of the brainstem characterized by yellowish, granular material associated with vessel hyperemia. On the floor of the fourth ventricle, there was deposition of yellowish, irregular, slightly granular material that protruded towards the obex and displaced the cerebellum dorsolaterally. Microscopically, there were pyogranulomatous, eosinophilic, necrotizing rhinitis and fibrinosuppurative, eosinophilic, necrotizing meningoencephalitis, both associated with thrombosis, vasculitis, and intralesional hyphae. The hyphae were impregnated with silver and presented thin, parallel walls, rarely septate and branched. At immunohistochemistry, the hyphae were immunostained with polyclonal anti-P. insidiosum antibody in fragments of the cerebellum and nasal cavity. The findings showed that P. insidiosum rhinitis can secondarily affect the nervous system of sheep, causing nonspecific neurological clinical signs.

Relata-se um caso de meningoencefalite por Pythium insidiosum secundária a rinite em uma ovelha mestiça, três anos de idade, proveniente de um rebanho de 15 animais, criados extensivamente e com acesso livre a açude. O animal apresentava dispneia leve, cegueira, midríase, opistótono, nistagmo, incoordenação, diminuição do tônus mandibular e espasticidade dos membros pélvicos. O exame macroscópico da cavidade nasal evidenciou uma massa vermelho-escura, irregular, friável, que comprometia bilateralmente o septo nasal e a porção rostral dos cornetos nasais. No encéfalo, havia espessamento difuso das leptomeninges do cerebelo e porção ventral do tronco encefálico caracterizado por material granular amarelado associado à hiperemia dos vasos. No assoalho do quarto ventrículo, havia deposição de material amarelado, irregular, levemente granuloso, que se projetava em direção ao óbex e deslocava o cerebelo dorsolateralmente. Microscopicamente, havia rinite piogranulomatosa, eosinofílica, necrosante e meningoencefalite fibrinossupurativa, eosinofílica, necrosante, ambas associadas à trombose, vasculite e hifas intralesionais. Pela utilização da técnica de GMS as hifas foram impregnadas pela prata e apresentavam paredes finas e paralelas, raramente septadas e ramificadas. Na imuno-histoquímica, houve imunomarcação com o anticorpo policlonal anti-P. insidiosum em fragmentos do cerebelo e cavidade nasal. Os achados evidenciaram que rinite por P. insidiosum pode afetar secundariamente o sistema nervoso de ovinos, causando sinais clínicos neurológicos inespecíficos.

Cutaneous pythiosis in equines in the Amazon Biome / Pitiose cutânea em equídeos no Bioma Amazônico

The study aimed to describe the clinicopathological aspects of 37 cases of pythiosis, 34 in horses and three in mules, from properties located in the Amazon biome of Pará, Brazil. The clinical signs observed in the animals were weakness, poor-to-regular nutritional status, pale mucous membranes, itching at the lesion site, and lameness when the limbs were affected. The lesions were located on the lips, nostrils, rib region, thoracic and abdominal walls, scapular, distal limbs, foreskin, perineum and udder. Macroscopically, ulcerative and granulation-tissue-like masses were observed, with fistulous tracts filled with yellowish and foul-smelling serosanguinous discharges. In the biopsy, it was possible to visualize white and firm areas with foci of yellowish necrotic material and hardened masses, called kunkers. The histopathological examination showed a pyogranulomatous inflammatory reaction with the presence of Pythium insidiosum hyphae, which were impregnated with black, confirming the diagnosis of pythiosis in equids in the Amazon biome, being the first report of the disease in mules in the region.

O trabalho objetivou descrever os aspectos clínico-patológicos de 37 casos suspeitos de pitiose, 34 em equinos e três em muares, provenientes de propriedades localizadas no Pará, bioma amazônico brasileiro. Os sinais clínicos observados nos animais eram caracterizados por debilidade, estado nutricional de ruim a regular, mucosas pálidas, prurido no local da lesão, além de claudicação quando os membros foram acometidos. As lesões eram localizadas nos lábios, narinas, região das costelas, parede torácica e abdominal, escapular, distais dos membros, prepúcio, períneo e úbere. Macroscopicamente observavam-se extensas lesões ulceradas com intensa proliferação de tecido de granulação, de bordos irregulares, com tratos fistulosos, de consistência firme, denominados de "kunkers", preenchidos com material amarelado e friável, possuindo exsudação serossanguinolenta de odor fétido. Ao exame histopatológico observou-se reação inflamatória piogranulomatosa com presença de hifas de Pythium insidiosum, as quais se impregnaram de negro, confirmando o diagnóstico de pitiose em equídeos no Bioma Amazônico, sendo o primeiro relato da doença em muares na região.

Canine gastrointestinal pythiosis: a case report in Brazilian Pantanal from a diagnostic approach / Pitiose gastrointestinal canina: relato de caso no Pantanal brasileiro a partir de uma abordagem diagnóstica

Pythiosis is caused by an aquatic fungus-like organism (Pythium insidiosum). It is considered an important public health issue as it can affect both animals and humans. This paper reports a case of gastrointestinal pythiosis in a dog. The patient was hospitalized for four days, during which the animal received supportive and symptomatic treatment. But the applied treatment was unsuccessful and the dog's clinical condition worsened, culminating in death. Complementary imaging tests such as radiography and ultrasonography, as well as hematological tests, were performed during the hospitalization period. The definitive diagnosis was reached in the postmortem as macroscopic and microscopic characteristics suggested the presence of intestinal granuloma and accentuated multifocal suppurative necrotic enteritis. Additionally, the histological evaluation revealed morphological structures compatible with P. insidiosum. Also, the results of nested PCR performed showed partial amplification (105 bp) of the ITS1 region of the ribosomal gene of P. insidiosum.(AU)

A pitiose é causada por um organismo aquático semelhante a um fungo (Pythium insidiosum) e considerada um importante problema de saúde pública, pois pode afetar animais e humanos. Este artigo relata um caso de pitiose gastrointestinal em um cão. O paciente ficou internado por quatro dias, período em que o animal recebeu tratamento de suporte e sintomático. No entanto, o tratamento aplicado não teve sucesso e o quadro clínico do cão piorou, culminando com a morte. Exames de imagem complementares, como radiografia e ultrassonografia, bem como exames hematológicos, foram realizados durante o período de internação. O diagnóstico definitivo foi feito na autópsia, pois as características macroscópicas e microscópicas sugeriam a presença de granuloma intestinal e acentuada enterite necrótica multifocal supurativa. Além disso, a avaliação histológica revelou estruturas morfológicas compatíveis com P. insidiosum. Além disso, a nested PCR foi realizada e mostrou amplificação parcial (105 pb) da região ITS1 do gene ribossomal de P. insidiosum.(AU)

Pitiose gástrica em canino jovem / Gastric pythiosis in a young canine

Pitiose gástrica canina é uma afecção crônica granulomatosa causada pelo pseudo-fungo Pythium insidiosum que ocasiona espessamento trasnsmural do estômago. Relata-se um caso de gastrite fúngica em um canino dobermann de 1 ano, macho, atendido no Hospital Veterinário da Universidade Federal do Mato Grosso, campus Sinop-MT, com sinais clínicos de diarreia escura intermitente, regurgitação, êmese e anorexia progressiva por cerca de três meses. No exame ultrassonográfico, observou espessamento gástrico difuso. Devido ao prognóstico ruim, foi realizado eutanásia. Na macroscopia, havia espessamento difuso da parede do estômago e petéquias multifocais por toda a mucosa. Na microscopia havia acentuado infiltrado inflamatório granulomatoso por toda a submucosa e muscular. Foi realizado coloração de metenamina nitrato de prata de Grocott (GMS), com evidenciação de hifas cenocíticas, raras septação e diâmetro de 6-10µm. A confirmação do diagnóstico e identificação do agente foi realizado através Imuno-histoquímica com anticorpo primário para Pythium insidiosum.

Canine gastric pythiosis is a chronic granulomatous infection caused by the insidious pythi fungus that causes stomach thickening. We report a case of fungal gastritis in a 1-year-old male progressive Doberman canine treated at the Veterinary Hospital of the Federal University of Mato Grosso, Sinop-MT campus, with clinical signs of intermittent diarrhea, regurgitation, vomiting and anorexia for about three months. On ultrasound examination, diffuse gastric thickening was observed. Due to the poor prognosis, euthanasia was performed. Macroscopically, there was diffuse thickening of the stomach wall and multifocal petechiae throughout the mucosa. On microscopy, there was a marked granulomatous inflammatory infiltrate throughout the submucosa and muscle Production of Grocott silver nitrate methenamine (GMS) was performed with evidence of cenocytic hyphae, rare septation and diameter of 6-10µm. Confirmation of the diagnosis and identification of the agent was performed through Immunohistochemistry with primary antibody to Pythium insidiosum.

Pitiose cutânea em equino no sertão do Ceará / Skin pythiosis in horse in the backwoods of the Ceará State

A pitiose é uma infecção invasiva ulcerativa piogranulomatosa causada pelo oomiceto Pythium insidiosum, parasita de plantas aquáticas em águas estagnadas. Apesar de não serem raros os relatos de pitiose nas espécies domésticas, a espécie equina é a mais afetada. A enfermidade pode ser adquirida através da colonização de lesões traumáticas e do folículo piloso. Em casos dessa infecção, pode-se encontrar hifas recobertas por células necróticas, formando massas branco-amareladas semelhantes a corais, denominadas de kunkers. As lesões são localizadas prioritariamente nas extremidades distais dos membros e na porção ventral da parede tóraco-abdominal. O diagnóstico da pitiose está relacionado a um prognóstico reservado dependendo do grau de comprometimento anatômico que a enfermidade se encontra. Diante do exposto, este trabalho teve como objetivo relatar um caso de pitiose cutânea em um equino, fêmea da raça Mangalarga Machador. O animal apresentava lesões ulcerativas no membro torácico esquerdo e no membro pélvico esquerdo. O tratamento escolhido foi o cirúrgico e imunoterápico, com intuito de remover todo o tecido lesionado e, posteriormente, coletar material para realização do exame histopatológico. O equino foi submetido a um protocolo pós-operatório com a terapia antitetânica por via intramuscular. A antibioticoterapia sistêmica foi realizada com penicilina benzatina, anti-inflamatório esteroidal, dexametasona foi realizada uma vez ao dia, durante cinco dias. Posteriormente, optou-se pelo uso do anti-inflamatório não esteroidal maxicam uma vez ao dia, durante cinco dias. O animal recebeu quatro doses do imunoterápico PITIUM-VAC por via subcutânea, com intervalo de 14 dias entre uma e outra aplicação, apresentando uma resposta satisfatória ao tratamento.

Pythiosis is an invasive ulcerative pyogranulomatous infection caused by the Oomycete Pythium insidiosum, a parasite of aquatic plants in standing water. Although reports of pythiosis in domestic species are not uncommon, equine species are the most affected. The disease can be acquired through the colonization of traumatic lesions and hair follicles. Hyphae covered by necrotic cells, forming yellowish-white coral-like masses called "kunkers" can be found. The lesions are located primarily on the distal extremities of the limbs and on the ventral portion of the thoracoabdominal wall. The diagnosis of pythiosis is linked to a poor prognosis, depending on the degree of anatomical involvement of the disease. Thus, this work aimed to report a case of cutaneous pythiosis in a female Mangalarga Machador equine. The animal presented ulcerative lesions on the left thoracic limb and the left pelvic limb. The chosen treatment was surgery and immunotherapy to remove all the injured tissue and, later, collect the material for histopathological examination. The horse was submitted to a post-operative protocol with intramuscular anti-tetanus therapy. Systemic antibiotic therapy with benzathine penicillin, steroidal anti-inflammatory, dexamethasone was performed once a day for five days. Subsequently, it was decided to use the non-steroidal anti-inflammatory maxicam once a day for five days. The animal received four doses of the immunotherapy PITIUM-VAC subcutaneously, with an interval of 14 days between applications, presenting a satisfactory response to the treatment.

Metastatic calcification and granulomatous grastroenteritis associated to Pythium insidiosum in a dog

Pythiosis is a granulomatous process of which the oomycete Pythium insidiosum is its etiological agent. It can affect animals and humans alike and its infection occurs when free zoospores in the water get in contact with the target tissues and encyst. The disease often occurs in tropical places with abundance of water and aquatic plants that host the fungus. Dogs infection is predominantly gastric with granuloma formations in the stomach and intestine with progressive signs of vomiting, weight loss and diarrhea. In this case report, we described clinical, surgical, necroscopic and histopathological findings of a one year and two months old, male boxer that presented clinical signs of anorexia and persistent vomiting. It was noticed on ultrasound examination an increase in stomach and intestine thickness. Laparotomy confirmed a mass affecting the gastric wall which, an incision biopsy, showed an abundant fibrous tissue associated with granulomatous reaction that was surrounded by tubuliform structures. Due to clinical complications, euthanasia was performed and in necroscopic examination a markedly increased stomach and duodenum was observed. An; histological examination of this areas it was observed that they contained granulation tissue with giant cells and epithelioids macrophages around necrosed areas associated with lymphocytes infiltrate. Also, it was possible to observe tubuliform structures by the Grocott-Gomori’s Methenamine Silver (GMS) stain, this finding is compatible with the agent Pythium insidiosum. Therefore, this presumptive identification was confirmed by PCR analysis which amplicon had 97.83% similarity with current available genomic sequence of P. insidiosum.

Metastatic calcification and granulomatous grastroenteritis associated to Pythium insidiosum in a dog

Pythiosis is a granulomatous process of which the oomycete Pythium insidiosum is its etiological agent. It can affect animals and humans alike and its infection occurs when free zoospores in the water get in contact with the target tissues and encyst. The disease often occurs in tropical places with abundance of water and aquatic plants that host the fungus. Dogs infection is predominantly gastric with granuloma formations in the stomach and intestine with progressive signs of vomiting, weight loss and diarrhea. In this case report, we described clinical, surgical, necroscopic and histopathological findings of a one year and two months old, male boxer that presented clinical signs of anorexia and persistent vomiting. It was noticed on ultrasound examination an increase in stomach and intestine thickness. Laparotomy confirmed a mass affecting the gastric wall which, an incision biopsy, showed an abundant fibrous tissue associated with granulomatous reaction that was surrounded by tubuliform structures. Due to clinical complications, euthanasia was performed and in necroscopic examination a markedly increased stomach and duodenum was observed. An; histological examination of this areas it was observed that they contained granulation tissue with giant cells and epithelioids macrophages around necrosed areas associated with lymphocytes infiltrate. Also, it was possible to observe tubuliform structures by the Grocott-Gomoris Methenamine Silver (GMS) stain, this finding is compatible with the agent Pythium insidiosum. Therefore, this presumptive identification was confirmed by PCR analysis which amplicon had 97.83% similarity with current available genomic sequence of P. insidiosum.(AU)

Cranioesophageal pythiosis in a horse

Background: Pythiosis is a chronic inflammatory disease that is caused by oomycete Pythium insidiosum. This illness affects several species including humans and horses. Equine is the most affected species, having no predisposition for breed, gender, or age. It is usually shown in cutaneous and subcutaneous forms, and the lesions, which grow quickly and are hard to treat, are located mainly in the extremities. The diagnosis is made via epidemiology, clinical signs, and macroscopic and microscopic aspects of the lesion. This study describes a case of cranioesophageal pythiosis in a horse, examining the epidemiological, clinical and pathological characteristics. Case: A 12-year-old male quarter horse, weighing 515 kg was taken to the Veterinary Hospital at the University Center of Espírito Santo (UNESC). The horse had an increase in volume in the cranioesophageal region, coughing, difficulty breathing, and a runny nose. On clinical examination, the horse showed an enlargement in the submandibular and retropharyngeal lymph nodes, subcutaneous edema in the larynx region, and a temperature of 38.2ºC. According to the owner, cough was recurrent and had lasted about 12 months even after treatment with different kinds of antimicrobials. On radiographic exam, there was a marked decrease in the tracheal lumen and increased soft tissue radiopacity in the region adjacent to the narrowing. The animal was taken to surgery to remove the mass, but he died because of complications during surgery. The animal’s owner did not allow necropsy, but a fragment of the mass in the cranioesophageal was removed and sent for histological examination. The fragment was fixed in 10% formalin and processed using routine histological analysis. Macroscopically, the mass was light yellowish and ulcerated, and it measured 7.0 × 5.0 × 5.0 cm. In the middle of the ulcerated areas, there were yellow and...

Cranioesophageal pythiosis in a horse

Background: Pythiosis is a chronic inflammatory disease that is caused by oomycete Pythium insidiosum. This illness affects several species including humans and horses. Equine is the most affected species, having no predisposition for breed, gender, or age. It is usually shown in cutaneous and subcutaneous forms, and the lesions, which grow quickly and are hard to treat, are located mainly in the extremities. The diagnosis is made via epidemiology, clinical signs, and macroscopic and microscopic aspects of the lesion. This study describes a case of cranioesophageal pythiosis in a horse, examining the epidemiological, clinical and pathological characteristics. Case: A 12-year-old male quarter horse, weighing 515 kg was taken to the Veterinary Hospital at the University Center of Espírito Santo (UNESC). The horse had an increase in volume in the cranioesophageal region, coughing, difficulty breathing, and a runny nose. On clinical examination, the horse showed an enlargement in the submandibular and retropharyngeal lymph nodes, subcutaneous edema in the larynx region, and a temperature of 38.2ºC. According to the owner, cough was recurrent and had lasted about 12 months even after treatment with different kinds of antimicrobials. On radiographic exam, there was a marked decrease in the tracheal lumen and increased soft tissue radiopacity in the region adjacent to the narrowing. The animal was taken to surgery to remove the mass, but he died because of complications during surgery. The animals owner did not allow necropsy, but a fragment of the mass in the cranioesophageal was removed and sent for histological examination. The fragment was fixed in 10% formalin and processed using routine histological analysis. Macroscopically, the mass was light yellowish and ulcerated, and it measured 7.0 × 5.0 × 5.0 cm. In the middle of the ulcerated areas, there were yellow and...(AU)

Pythiosis in cattle in Northeastern Brazil / Pitiose em bovinos no Nordeste do Brasil

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)

Pythiosis in cattle in Northeastern Brazil / Pitiose em bovinos no Nordeste do Brasil

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)

Granulomatous pneumonia due to pythiosis in captive South American coatis (Nasua nasua) / Pneumonia granulomatosa decorrente de pitiose em quatis sul-americanos (Nasua nasua) de cativeiro

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)

Granulomatous pneumonia due to pythiosis in captive South American coatis (Nasua nasua) / Pneumonia granulomatosa decorrente de pitiose em quatis sul-americanos (Nasua nasua) de cativeiro

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)

Segmental enteritis associated with Pythium insidiosum infection in a horse

Background: Pythiosis is an infectious disease caused by the oomycete Pythium insidiosum, with higher occurrence in wetlands and hot climate regions. This microorganism develops its cycle in aquatic plants, and most cases happen because of the contact of animals or people with water containing the motile zoospores (infectious form). Horses are the principal species affected and develop principally cutaneous and subcutaneous lesions, but the gastrointestinal tract is seldom affected. Humans develop various forms of pythiosis, such as a vascular form. The objectives of the current study are to describe an unusual case of intestinal pythiosis, its clinical signs, aspects of pathogenesis, and diagnosis. Case: A 13-year-old Crioula mare, from Santa Maria, RS, Brazil, presented with reduced food and water intake, apathy, restlessness, rolling, nasal reflux, firm abdomen upon palpation, and tachypnea during 2 days. The horses of this farm were fed native pasture and horse feed, and they had access to a nearby pond. Two days following the start of the clinical signs, the horse died and was necropsied on the farm. During necropsy, there was around 400 mL of reddish effusion in the abdominal cavity (modified transudate). A 15 cm segment of jejunum was firm upon palpation and had a severe transmural thickening. The wall of the affected area was up to 3 cm in thickness and firm, with small yellowish and irregular masses that stood out and looked friable, interpreted as kunkers. Microscopically, the yellowish masses (kunkers) were characterized by dense accumulations of intact and degenerate eosinophils (eosinophilic necrosis). Within these kunkers, and also on their periphery, there were multiple negatively stained hyphal profiles. Hyphae were also seen on the wall of small arteries inside the kunkers. These hyphae had nearly parallel walls and were occasionally...

Segmental enteritis associated with Pythium insidiosum infection in a horse

Background: Pythiosis is an infectious disease caused by the oomycete Pythium insidiosum, with higher occurrence in wetlands and hot climate regions. This microorganism develops its cycle in aquatic plants, and most cases happen because of the contact of animals or people with water containing the motile zoospores (infectious form). Horses are the principal species affected and develop principally cutaneous and subcutaneous lesions, but the gastrointestinal tract is seldom affected. Humans develop various forms of pythiosis, such as a vascular form. The objectives of the current study are to describe an unusual case of intestinal pythiosis, its clinical signs, aspects of pathogenesis, and diagnosis. Case: A 13-year-old Crioula mare, from Santa Maria, RS, Brazil, presented with reduced food and water intake, apathy, restlessness, rolling, nasal reflux, firm abdomen upon palpation, and tachypnea during 2 days. The horses of this farm were fed native pasture and horse feed, and they had access to a nearby pond. Two days following the start of the clinical signs, the horse died and was necropsied on the farm. During necropsy, there was around 400 mL of reddish effusion in the abdominal cavity (modified transudate). A 15 cm segment of jejunum was firm upon palpation and had a severe transmural thickening. The wall of the affected area was up to 3 cm in thickness and firm, with small yellowish and irregular masses that stood out and looked friable, interpreted as kunkers. Microscopically, the yellowish masses (kunkers) were characterized by dense accumulations of intact and degenerate eosinophils (eosinophilic necrosis). Within these kunkers, and also on their periphery, there were multiple negatively stained hyphal profiles. Hyphae were also seen on the wall of small arteries inside the kunkers. These hyphae had nearly parallel walls and were occasionally...(AU)

Genotyping of South American clinical isolates of Pythium insidiosum based on single nucleotide polymorphism-based multiplex PCR / Genotipagem de isolados clínicos de Pythium insidiosum da América do Sul utilizando polimorfismos de nucleotídeo único baseado em PCR multiplex

We aimed to genotype the South American clinical isolates of Pythium insidiosum using the single nucleotide polymorphisms (SNP) of the ribosomal DNA sequences (rDNA). Previously, an SNP-based multiplex-PCR was able to distinguish three different clades of P. insidiosum isolates. Thus, we used this assay to evaluate South American clinical isolates of P. insidiosum (n=32), standard strains from Costa Rica (n=4), Thailand (n=3), Japan (n=1), and India (n=1), a standard strain of Pythium aphanidermatum, and Brazilian environmental isolates of Pythium torulosum, Pythium rhizo-oryzae and Pythium pachycaule voucher (n=3). It was possible to allocate each American P. insidiosum isolate to clade I, the isolates of India, Japan, and Thailand to clade II, and the Thai isolate to clade III. P. aphanidermatum, P.torulosum, P.rhizo-oryzae and P.pachycaule voucher isolates were not amplified. For the first time, a P. insidiosum isolate from Uruguay, South America, was included in molecular analyzes. By SNP-based multiplex-PCR, it was possible to perform the identification and genotyping of the South American isolates of P. insidiosum, demonstrating similar genetic characteristics of these isolates.(AU)

O objetivo deste estudo foi genotipar isolados clínicos de Pythium insidiosum da América do Sul utilizando polimorfismos de nucleotídeo único (SNP) de sequências de rDNA. Anteriormente, um multiplex-PCR baseado em SNP foi capaz de distinguir P. insidiosum em três diferentes clados. Dessa forma, utilizamos este método para avaliar isolados clínicos de P. insidiosum da América do Sul (n=32), cepas padrão da Costa Rica (n=4), Tailândia (n=3), Japão (n=1) e Índia (n=1), uma cepa padrão de Pythium aphanidermatum e isolados ambientais brasileiros de Pythium torulosum; Pythium rhizo-oryzae e Pythium pachycaule voucher (n=3). Os isolados analisados foram alocados aos clados: I (americanos), II (isolados da Índia, Japão e Tailândia), e III (um isolado tailandês). P. aphanidermatum, P.torulosum, P.rhizo-oryzae e P.pachycaule voucher não foram amplificados. Pela primeira vez, um isolado de P. insidiosum do Uruguai foi incluído em análises moleculares. Através da multiplex-PCR baseada em SNP, foi possível realizar a identificação e genotipagem dos isolados sul-americanos de P. insidiosum, demonstrando características genéticas semelhantes entre esses isolados.(AU)

Equine pythiosis in the eastern wetlands of Uruguay / Pitiose equina em áreas úmidas do leste do Uruguai

Equine pythiosis is an ulcerative and granulomatous disease of the skin, caused by the oomycete Pythium insidiosum (Pythiaceae). The objective of this study was to describe seven cases of equine pythiosis that occurred from 2012 to 2017 in the eastern region of Uruguay. Six of the seven cases occurred in the eastern wetland ecosystems of the Merin basin, and the remaining case occurred in the wetland fluvial plains of the Tacuarembó River. Lesions consisted of a large, rapidly growing ulcerated tumor with abundant granulation tissue, serosanguineous secretion, and fistulous tracts containing large concretions or kunkers. The animals presented intense pruritus, claudication and loss of body condition, with death or euthanasia in extremis in six cases. The main histological lesions consisted of an eosinophilic and pyogranulomatous inflammatory process, with numerous foci of eosinophilic necrosis (kunkers), collagenolysis, and a Splendore-Hoeppli reaction. In all cases, silver coloration (Grocott) showed intralesional hyphae compatible with P. insidiosum, which was confirmed by immunohistochemistry in three cases. A horse in the terminal phase of the disease was treated with triamcinolone acetonide (50mg IM every 15 days), and fully recovered after 1 year. It is concluded that equine pythiosis is prevalent in the wetland ecosystems of eastern Uruguay and that treatment with triamcinolone is auspicious.(AU)

Pitiose é uma doença granulomatosa e ulcerativa da pele dos equinos causada pelo oomyceto Pythium insidiosum (Pythiaceae). O objetivo deste trabalho foi descrever sete casos de pitiose equina que ocorreram de 2012 a 2017 na região leste do Uruguai. Seis dos sete casos ocorreram no ecossistema de áreas pantanosas da bacia da bacia da Lagoa Mirim Merin e o restante nas planícies fluviais pantanosas do rio Tacuarembó. As lesões se caracterizaram por tumores ulcerados de crescimento rápido com abundante tecido de granulação, secreção serossanguinolenta e presença de tratos fistulosos contendo material coraloide ou kunkers. Os equinos apresentavam prurido intenso, claudicação e perda da condição corporal e seis morreram ou foram eutanasiados in extremis. As principais lesões histológicas consistiam de um processo inflamatório piogranulomatoso com numerosos focos de necrose eosinofílicos (kunkers), colagenólise e reação de Splendori-Hoepli. Em todos os casos a impregnação pela prata (Grocott) revelou a presença de hifas intralesionais compatíveis com P. insidiosum, o que foi confirmado pela imuno-histoquímica em três casos. Um equino em fase terminal da doença foi tratado com triamcinolona acetonida (50mg, IM, a cada 15 dias), recuperando-se completamente após um ano. Conclui-se que a pitiose é uma enfermidade presente em áreas úmidas na região leste do Uruguai e o tratamento com triamcinolona pode ser uma alternativa promissora.(AU)

Equine pythiosis in the eastern wetlands of Uruguay / Pitiose equina em áreas úmidas do leste do Uruguai

Equine pythiosis is an ulcerative and granulomatous disease of the skin, caused by the oomycete Pythium insidiosum (Pythiaceae). The objective of this study was to describe seven cases of equine pythiosis that occurred from 2012 to 2017 in the eastern region of Uruguay. Six of the seven cases occurred in the eastern wetland ecosystems of the Merin basin, and the remaining case occurred in the wetland fluvial plains of the Tacuarembó River. Lesions consisted of a large, rapidly growing ulcerated tumor with abundant granulation tissue, serosanguineous secretion, and fistulous tracts containing large concretions or kunkers. The animals presented intense pruritus, claudication and loss of body condition, with death or euthanasia in extremis in six cases. The main histological lesions consisted of an eosinophilic and pyogranulomatous inflammatory process, with numerous foci of eosinophilic necrosis (kunkers), collagenolysis, and a Splendore-Hoeppli reaction. In all cases, silver coloration (Grocott) showed intralesional hyphae compatible with P. insidiosum, which was confirmed by immunohistochemistry in three cases. A horse in the terminal phase of the disease was treated with triamcinolone acetonide (50mg IM every 15 days), and fully recovered after 1 year. It is concluded that equine pythiosis is prevalent in the wetland ecosystems of eastern Uruguay and that treatment with triamcinolone is auspicious.(AU)

Pitiose é uma doença granulomatosa e ulcerativa da pele dos equinos causada pelo oomyceto Pythium insidiosum (Pythiaceae). O objetivo deste trabalho foi descrever sete casos de pitiose equina que ocorreram de 2012 a 2017 na região leste do Uruguai. Seis dos sete casos ocorreram no ecossistema de áreas pantanosas da bacia da bacia da Lagoa Mirim Merin e o restante nas planícies fluviais pantanosas do rio Tacuarembó. As lesões se caracterizaram por tumores ulcerados de crescimento rápido com abundante tecido de granulação, secreção serossanguinolenta e presença de tratos fistulosos contendo material coraloide ou kunkers. Os equinos apresentavam prurido intenso, claudicação e perda da condição corporal e seis morreram ou foram eutanasiados in extremis. As principais lesões histológicas consistiam de um processo inflamatório piogranulomatoso com numerosos focos de necrose eosinofílicos (kunkers), colagenólise e reação de Splendori-Hoepli. Em todos os casos a impregnação pela prata (Grocott) revelou a presença de hifas intralesionais compatíveis com P. insidiosum, o que foi confirmado pela imuno-histoquímica em três casos. Um equino em fase terminal da doença foi tratado com triamcinolona acetonida (50mg, IM, a cada 15 dias), recuperando-se completamente após um ano. Conclui-se que a pitiose é uma enfermidade presente em áreas úmidas na região leste do Uruguai e o tratamento com triamcinolona pode ser uma alternativa promissora.(AU)