Outcomes of therapeutic penetrating keratoplasty in Pythium insidiosum keratitis managed with a combination of antibiotics

Purpose: To describe the clinical outcomes of therapeutic penetrating keratoplasty (TPK) in patients with Pythium insidiosum keratitis following treatment with anti?pythium therapy (APT) consisting of linezolid and azithromycin. Methods: A retrospective review of medical records from May 2016 to December 2019 of patients with P. insidiosum keratitis was carried out. Patients who were treated with APT for a minimum of 2 weeks and then subsequently underwent TPK were included in the study. Data on demographic characteristics, clinical features, microbiology characteristics, and intraoperative details, postoperative outcomes were documented. Results: A total of 238 cases of Pythium keratitis were seen during the study period and 50 cases that satisfied the inclusion criteria were included. The median of the geometric mean of the infiltrate was 5.6 mm (IQR 4.0–7.2 mm). The patients received topical APT for a median of 35 days (IQR 25–56) prior to surgery. The most common indication of TPK was worsening keratitis (41/50, 82%). No recurrence of infection was observed. An anatomically stable globe was noted in 49/50 eyes (98%). The median graft survival rate was 2.4 months. A clear graft was present in 10 eyes (20%) with a final median visual acuity of 20/125 after a median follow?up period of 18.4 months (IQR 11–26 months). Graft size of less than 10 mm [OR: 5.824 (CI:1.292?41.6), P = 0.02] was found to be significantly associated with a clear graft. Conclusion: Performing TPK following the administration of APT has good anatomical outcomes. A smaller graft of <10 mm was associated with a higher chance of graft survival

Clinical differentiation of keratitis due to fungus and Pythium: A photographic survey

Purpose: To evaluate the knowledge and skills of clinicians in differentiating cases of Pythium from fungal keratitis based on their clinical appearance. Methods: Thirty slit?lamp photographs from culture?proven cases of Pythium and fungal keratitis were selected for conducting a prospective online?based photographic survey. The participants were asked to identify the causative organism and enlist the specific clinical features which helped in reaching the diagnosis. Both cornea specialists and clinical fellows/trainees participated in the survey. Sensitivity, specificity, and predictive values were estimated, and the factors associated with these, including the identification rates of clinical signs, were evaluated using bivariate and multivariate linear regression analyses. The identification rates between the consultants and fellows were compared. Results: A total of 42 cornea specialists (28 consultants and 14 clinical fellows) participated in the survey. Clinicians were able to differentiate Pythium from fungal etiology in only 56% of cases. The differentiating ability between the consultants and fellows was statistically insignificant. Average sensitivity, specificity, and positive and negative predictive values for differentiating Pythium from fungus were 56%, 65%, 56%, and 66%, respectively. The specific clinical features enlisted by the participants to differentiate Pythium from fungus were the presence of tentacles, peripheral guttering, dot?like infiltrates, and elevated plaques. Conclusion: Clinical knowledge regarding the signs and symptoms and the microbiological identification of features for Pythium are still lacking among clinicians. As the treatment protocols are distinctly different for both, it is imperative to enhance the knowledge and diagnosing ability to tackle this emerging microorganism which causes high morbidity

Research progress of pythiosis in human / 中华临床感染病杂志

Pythiosis is a rare and refractory infectious disease in human caused by Pythium insidiosum, which mainly occurs in the tropics, subtropics and some temperate regions. There are few reports of pythiosis in China. The mortality of the disease is very high, however, there is uncertainty in its diagnosis and treatment. This article reviews the epidemiological characteristics, clinical manifestations, diagnostic methods, treatment strategies, and prognosis of pythiosis in human, to provide reference for clinical management of pythiosis.

Clinical differentiation of Pythium keratitis from fungal keratitis and development of a scoring system

Purpose: To differentiate Pythium keratitis from fungal keratitis using clinical signs, to explore usefulness of various signs as diagnostic prognosticators, and develop a clinical scoring system. Methods: A retrospective review of medical records and archived clinical photographs of patients with culture?positive Pythium keratitis and hyaline filamentous fungal keratitis was conducted at a tertiary eye institute to explore characteristics of ulcers that may aid diagnosis. Results: Full?thickness corneal stromal keratitis (P = 0.055), a dry ulcer surface (P = 0.010), tentacles (P < 0.0001), intrastromal dots (P < 0.0001), ring infiltrates (P = 0.024), reticular patterns (P < 0.0001), and peripheral furrows (P < 0.0001) were clinical signs associated with Pythium keratitis. Multiple regression analysis identified tentacles (odds ratio: 24.1, 95% confidence interval (CI): 3.8–158.1, P = 0.001) and peripheral furrows (odds ratio: 60.6, 95% CI: 5.1–712.3, P = 0.001) as independent diagnostic prognosticators for Pythium keratitis. The positive and negative likelihood ratios of a dry ulcer surface, tentacles, intrastromal dots, ring infiltrates, reticular patterns, and peripheral furrows predicting Pythium keratitis were 1.6, 13.6, 17.9, 4.3, 30.7, 15.3 and 0.4, 0.4, 0.7, 0.9, 0.6 and 0.8, respectively. The presence of two or more of these clinical signs (excluding a dry ulcer surface) had a sensitivity of 55.6% and a false positive rate of 1.4%. Conclusion: Tentacles, intrastromal dots, ring infiltrates, reticular patterns, and peripheral furrows are clinical signs to be considered for the diagnosis of Pythium keratitis and the presence of two or more signs has a very low false positive rate

Sensitivity and specificity of potassium hydroxide and calcofluor white stain to differentiate between fungal and Pythium filaments in corneal scrapings from patients of Pythium keratitis

Purpose: To assess the sensitivity of potassium hydroxide and calcofluor white (KOH+CFW) mount in the diagnosis of Pythium keratitis and concordance among microbiologists. Methods: Three microbiologists evaluated the microscopic images of KOH + CFW mounts of confirmed cases of Pythium and fungal keratitis seen between January 2019 and February 2021. The filaments were compared using specific differentiating features. The sensitivity and specificity of KOH + CFW in diagnosing Pythium infection were evaluated along with concordance among the microbiologists. Results: Sixty consecutive cases with confirmed growth of fungus or Pythium insidiosum (n = 29) were evaluated. The sensitivity of KOH + CFW in the correct identification of Pythium filaments ranged from 79.3% to 96.5% among three microbiologists. There was good interobserver (k = 0.76–0.90) and intraobserver (k = 0.70–0.97) agreements among three microbiologists. The differentiating findings (P < 0.0001) suggestive of Pythium filaments were the absence of septae in 23 (79.3%) and collapsed walls in 22 (75.9%) cases. Conclusion: KOH + CFW has good sensitivity and specificity in the diagnosis of Pythium keratitis with good interobserver and intraobserver concordance.

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)

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)

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)

Bio-efficacy of Bacillus subtilis against Damping off Disease in Brinjal

Bacillus sp is one of the biocontrol agents are extensively used in management of fungal diseases of crop plants, exhibiting mycoparasitism against a wide range of plant pathogens. In the present investigation Efficacy of Bacillus subtilis was tested against Pythium aphanidermatum under glass house and field conditions. With regard to the germination and seedling growth parameters, the treatment T2 (liquid formulation of Bacillus subtilis @10 ml/L) recorded the highest germination percentage (92.59%), shoot length (59.3 cm), root length (24.3 cm), vigor index (7740.52) and yield (22.67 mt/ha). This was followed by the treatments T3, T1 and T4 in the decreasing order of merit. In the biometric observations also, 10 ml/L and 20 ml/L concentration of liquid formulation of Bacillus subtilis recorded statistically significant results. Observations on total protein content in brinjal plants treated with different concentrations of Bacillus subtilis revealed that treatment T2 was found best in both field and glass house conditions yielded highest protein as 0.44mg/ml and 0.30mg/ml respectively. The least values of germination, growth parameters, protein content and yield were observed in untreated control.

Pythium keratitis in South India: Incidence, clinical profile, management, and treatment recommendation

Purpose: To study the demographic profile, clinical features, treatment outcome, and ocular morbidity of microbiologically proven Pythium keratitis in South India. Methods: A retrospective analysis of clinical records of microbiologically proven Pythium keratitis at a tertiary eye care referral center in South India from January 2016 to November 2017 was performed. Demographic details, predisposing risk factors, microbiological investigations, clinical course, and visual outcome were analyzed. Results: Seventy-one patients with microbiologically proven Pythium keratitis were identified. The mean age was 44(±18.2) years with an increase in male preponderance and 50% were farmers. Duration of delay at time of presentation to the hospital was a mean of 14(±7.2) days. The visual acuity at baseline ranged from 6/6 to no light perception (median 2.1 logMAR). A combination of 5% natamycin and 1% voriconazole was given to 42% patients, and natamycin alone was given to 39.4% patients. 1% itraconazole eye drops alone was initiated in 7 (10%) patients and 3 among this group responded. Therapeutic keratoplasty (TPK) was performed in 48 (67.6%) patients. None of the primary grafts remained clear after a period of 1 month. Twenty-six eyes (54.2%) had graft reinfection and all these eyes either developed anterior staphyloma (4) or were eviscerated (3) and 13 eyes became phthisical. The remaining 22 patients who had TPK resulted in failed graft. Among these, re-grafts were performed in 6 patients, of which 5 were doing well at the last follow-up. Conclusion: We report a large series of patients with Pythium keratitis. Promoting early and differential diagnosis, awareness of clinicians and specific treatment options are needed for this devastating corneal disease.

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

ABSTRACT: 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.

RESUMO: 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.

Feline subcutaneous pythiosis / Pitiose subcutânea em um felino

ABSTRACT: Pythiosis in felines is a rare disease associated with the oomycete Pythium insidiosum. The aim of this report was to describe the macroscopic, histopathological, and molecular characteristics of P. insidiosum infection in a 2-year-old cat, with a localized invasive subcutaneous mass. The feline had an increase of volume near the anal region since it was younger. The cat died just after surgery. The necropsy was performed, and samples were collected for histopathological examination. Microscopically, the skin lesion was characterized by necro-eosinophilic dermatitis, panniculitis, and myositis surrounding negatively stained hyphal structures. In the sections stained with GMS, dark brown hyphae were clearly seen inside the affected tissue. They were rarely septate and their walls were almost parallel. Immunohistochemistry using a polyclonal anti-P. insidiosum antibody showed a strongly immunostained hyphae into the lesions. The analysis based on PCR had a positive result for P. insidiosum. Pythiosis should be considered in the differential diagnosis of subcutaneous tissue disorders in felines.

RESUMO: Pitiose em felinos é uma doença de ocorrência rara associada ao oomiceto Pythium insidiosum. O objetivo deste relato é descrever as características macroscópicas, histopatológicas e moleculares da infecção por P. insidiosum em um gato de dois anos de idade, com uma massa invasiva localizada no subcutâneo. O gato morreu logo após a cirurgia, sendo realizada a necropsia e coleta de amostras para exame histopatológico. Microscopicamente, a lesão cutânea foi caracterizada por dermatite necroeosinofílica, paniculite e miosite envolvendo imagens de hifas negativamente coradas. Nas seções coradas com GMS, hifas marrom-escuras foram claramente vistas dentro do tecido afetado. As hifas raramente eram septadas e suas paredes eram quase paralelas. A imuno-histoquímica, utilizando um anticorpo policlonal anti-P. insidiosum, mostrou hifas fortemente imunomarcadas nas lesões. A análise baseada em PCR teve resultado positivo para P. insidiosum. A pitiose deve ser considerada no diagnóstico diferencial de desordens teciduais subcutâneas em felinos.

Pythiosis with atypical location in the soft palate in a horse in Southern Brazil / Pitiose com localização atípica no palato mole em um equino na região Sul do Brasil

A pitiose é uma enfermidade piogranulomatosa causada pelo oomiceto aquático Pythium insidiosum, que acomete várias espécies domésticas, assim como a humana. O presente relato refere-se à descrição de um caso de pitiose em um equino, fêmea, da raça Crioula, criado na região Sul do Brasil com história clínica de emagrecimento progressivo, disfagia e dificuldade respiratória. Macroscopicamente, o palato mole estava aumentado de tamanho e deslocado caudalmente, resultando em obstrução física da nasofaringe. Ao corte, essa massa era branca-amarelada, fibrosa e com áreas de ulceração. Na análise histopatológica, havia inflamação piogranulomatosa, com focos necróticos associados a imagens negativas tubuliformes de hifas fúngicas. A coloração de Grocott demonstrou inúmeras hifas septadas e ramificadas, as quais foram marcadas positivamente para anticorpos contra P. insidiosum na imuno-histoquímica. O diagnóstico de pitiose foi baseado nos achados macro e microscópicos e confirmado pela técnica de imuno-histoquímica.(AU)

Pitiose equina: revisão de literatura / Equine phityosis: literature review / Pitiosis equina: revisión de literatura

A pitiose é uma enfermidade que afeta diversas espécies animais, sendo a espécie equina geralmente a mais acometida. É causada por um oomiceto que se desenvolve principalmente em regiões subtropicais e tropicais em áreas alagadiças. As lesões frequentemente são únicas, bastante exsudativas, com prurido moderado a intenso e tendem a se localizar nas regiões do corpo que mais entram em contato com as áreas alagadas. Os meios de diagnostico e tratamento vêm sendo estudados surgindo assim cada vez mais técnicas e alternativas. Por ser uma doença que pode causar diversos impactos econômicos, seja por gastos com o tratamento ou até perda da função ou morte do animal, é uma doença que merece atenção no mercado de equinos no Brasil e no mundo. Este trabalho tem por objetivo abordar as principais características da pitiose, seu diagnóstico, tratamento e possíveis impactos.(AU)

Pythiosis is a disorder affecting several animal species; equines are usually the most widely affected species. The disease is caused by oomycete mainly developed in swampy areas in tropical and subtropical regions. The lesions are frequently single, exudative wounds, with moderate to intense itching, with a tendency to be located in parts that are in contact with wetlands. Diagnostic and treatment procedures have been studied, resulting in increasingly techniques and alternatives. This condition causes several economic impacts, either by expenditure with treatment, loss of function or even death of the animal. Therefore, greater care in the equine market must be provided, both in Brazil and worldwide. This study aims to describe the main features of pythiosis, its diagnosis, treatment and possible impacts.(AU)

Pitiosis es una enfermedad que afecta a varias especies animales, siendo la espécie equina generalmente la más afectada. Es causada por un oomiceto que se desarrolla principalmente en las regiones tropicales y subtropicales en las zonas pantanosas. Las lesiones, con frecuencia son únicas, exudativas, con prurito moderado a intenso, y tienden a ubicarse en regiones del cuerpo que más entran en contacto com zonas húmedas. Los procedimientos de diagnóstico y tratamento vienen siendo estudiados, lo que resulta cada vez más en técnicas y alternativas. Por ser una enfermedad que puede causar varios impactos económicos, sea por gastos con el tratamiento, la pérdida de la función o la muerte del animal, es una enfermedad que merece atención en el mercado de equinos en Brasil y en el mundo. Este estudio há tenido como objetivo abordar las principales características de pitiosis, su diagnóstico, tratamiento y los posibles impactos.(AU)

Pythiosis cutaneous in horses treated with triamcinolone acetonide. Part 3. Histomorphometric analysis / Pythiosis cutánea en equinos tratados con acetonida de triamcinolona. Parte 3. Análisis histomorfométrico

ABSTRACT Objective. The objective of the study was to analyze Histomorphometrically of the healing process with cutaneous pythiosis in horses treated with triamcinolone acetonide. Materials and methods. 24 horses with pythiosis were used, to a group 50 mg of intramuscular triamcinolone acetonide (GT) was applied, while the other group was not applied treatment (GC). They were collected tissue biopsies, processed, sliced and stained with hematoxylin and eosin (H & E), Gomori trichrome (TG), picrosirius red / polarization (PR / P) and Grocott methenamine silver (GMS). Photomicrographs were selected and 10 histological changes, analyzed with BioEstat 5.0 software, obtaining quantities of tissue cells such as eosinophils, neutrophils, macrophages, fibroblasts and collagen through planimetric evaluation point count. Results. In GSM staining was observed decrease in the presence of intralesional hyphae of P. insidiosum to 16 days (p<0.05). Staining H&E, we observed a decrease of the inflammatory process, shown in eosinophils (p=0.0001), neutrophils (p=0.0001), and macrophage (p=0.00001). In the staining of GT and PR/P increase the amount of fibroblasts and collagen fibers were observed, also the gradual exchange of type III collagen to type I, increased fibroblast show significant (p=0.0001) from day 16 until day 40, the expression of collagen was significant (p=0.0001) from day 16 until the end of the study. It was statistically significant correlation between neutrophils and macrophages (p=0.00018), collagen and eosinophil (p=0.03) and fibroblasts and collagen (p=0.02). The animals in the CG do not present histomorphometric improvement during the study. Conclusions. We conclude that the cell produces triamcinolone acetonide and histomorphometric tecidual recovery in horses with pythiosis.

RESUMEN Objetivo. El objetivo del estudio fue analizar histomorfométricamente el proceso de cicatrización en equinos con pythiosis tratados con acetonida de triamcinolona. Materiales y métodos. Se utilizaron 24 equinos con pythiosis, a un grupo se aplicó 50 mg de acetonida de triamcinolona intramuscular (GT), mientras que al otro grupo no fue aplicado tratamiento (GC). Fueron colectadas biopsias de tejido, procesadas, laminadas y coloreadas con hematoxilina eosina (H&E), tricrómico de Gómori (TG), picrosirius red/polarization (PR/P) y plata metanamina de Grocott (GMS). Fueron seleccionados y fotomicrografiados 10 campos histológicos, analizados con el software BioEstat 5.0, obteniéndose las cantidades tisulares de células como eosinófilos, neutrófilos, macrófagos, fibroblastos y colágeno a través de evaluación planimétrica por conteo de puntos. Resultados. En coloración de GSM, se observó disminución de hifas intralesionales del Pythium a los 16 días (p<0.05). En coloración de H&E, hubo disminución del proceso inflamatorio, evidenciado en eosinófilos (p=0.0001), neutrófilos (p=0,0001) y macrófagos (p=0.00001). En coloraciones de TG y PR/P, hubo aumento de fibroblastos, fibras colágenas y cambio progresivo de colágeno tipo III a tipo I, el aumento de fibroblastos fue significativo (p=0.0001) del día 16 al 40, la expresión de colágeno fue altamente significativa (p=0.0001) del día 16 hasta el final del estudio. Hubo correlación significativa entre neutrófilos y macrófagos (p=0.00018), colágeno y eosinófilos (p=0.03) y colágeno y fibroblastos (p=0.02). Los animales del GC no presentaron mejoría histomorfométrica durante el estudio. Conclusiones. Es posible concluir que la acetonida de triamcinolona produce recuperación histomorfométrica celular y tecidual en los equinos con pythiosis.

Intestinal intussusception secondary to enteritis caused by Pythium insidiosum in a bitch: case report / Intussuscepção intestinal secundária à enterite causada por Pythium insidiosum em uma cadela: relato de caso

We report a case of intestinal intussusception secondary to enteritis caused by Pythium insidiosum infection in a 1-year-old mixed breed bitch that died after a 30-day history of bloody diarrhea, anorexia, weight loss, listlessness, dehydration, and pale mucous membranes. Necropsy revealed two areas of intussusception, one jejunum-jejunal and one ileum-cecum-colic. The first intussusception showed slight congestion of the intestinal loop, which was resolved by manual traction, while the second intussusception exhibited thickening, irregular yellow spots, and transmural congestion. Histologically, the jejunum-jejunal segment had ischemic lesions secondary and ileum-cecum-colic intussusception there was also necrotizing, pyogranulomatous enteritis associated with negative images of intralesional fungal hyphae that were well visualized by silver impregnation. Immunohistochemistry identified the hyphae as Pythium insidiosum. The diagnosis was made based on the histological changes and confirmed by immunohistochemistry. We can conclude that enteritis by P. insidiosum in dogs can occasionally cause intestinal intussusception and result in acute death.(AU)

Relata-se um caso de intussuscepção intestinal secundária à enterite causada por Pythium insidiosusm em uma cadela, sem raça definida, com um ano de idade, que morreu após apresentar 30 dias de diarreia sanguinolenta, anorexia, perda de peso, apatia, desidratação e mucosas pálidas. Na necropsia verificaram-se duas áreas de intussuscepções, sendo uma jejuno-jejunal e outra íleo-ceco-cólica. Na primeira havia discreta congestão, que foi desfeita pela tração manual, e na segunda observou-se espessamento segmentar, focos amarelados irregulares e congestão transmural do intestino. Microscopicamente, no segmento jejuno-jejunal observaram-se moderadas lesões isquêmicas secundárias à intussuscepção, e no segmento íleo-ceco-cólica verificou-se enterite piogranulomatosa e necrosante, segmentar, associada a imagens negativas de hifas intralesionais, impregnadas pela prata. Pela imuno-histoquímica, as hifas foram imunomarcadas para Pythium insidiosum. O diagnóstico foi realizado com base nas características microscópicas das lesões e morfotintoriais das hifas e foi confirmado pela imuno-histoquímica. De acordo com esses dados, é possível concluir que a infecção intestinal por P. insidiosum em cães pode causar intussuscepção e morte.(AU)

Distribución geográfica de la casuística clínica bovina del servicio ambulatorio de grandes animales de la Universidad de Córdoba (Colombia) / Geographic distribution of bovine clinical casuistry in the outpatient service for large animals at the Universidad de Córdoba (Colombia) / Distribuição geográfica da casuística clínica bovina do serviço ambulatório de grandes animais da Universidade de Córdoba (Colômbia)

Resumen En este estudio se realizó la distribución geográfica de la casuística clínica bovina del servicio ambulatorio de grandes animales de la Universidad de Córdoba (Colombia). Fueron consideradas dentro del estudio solo aquellas historias clínicas completas que presentaran un diagnóstico definitivo. Los casos atendidos fueron clasificados según el sistema afectado, el diagnóstico definitivo, el tipo de tratamiento (médico o quirúrgico) y la ubicación geográfica en Córdoba. Se analizaron 6356 historias clínicas de casos presentados entre 2005 y 2016, de las cuales el 77,09 % fueron en la especie bovina, y el sistema más afectado fue el hematopoyético con 79,29 % de la casuística, seguido del sistema tegumentario (11,18 %) y el sistema digestivo (2,86 %). Se atendió el 73 % de los municipios del departamento, donde se observó un alto predominio de alteraciones en el sistema hematopoyético (79,29 %), seguido por el sistema tegumentario (11,18 %) y el sistema digestivo (2,86 %). Se estableció que el sistema hematopoyético fue afectado por hematrópicos endoglobulares como Babesia spp. y Anaplasma spp., mientras que el sistema tegumentario fue afectado principalmente por infestaciones con Rhabditis spp. (41 %), seguido de infecciones por dermatofitosis (19 %) y papilomavirus (16 %). Se puede concluir que el servicio ambulatorio de la Clínica Médico-Quirúrgica de Grandes Animales de la Universidad de Córdoba posee una amplia y variada casuística, lo que se demuestra variedad en las frecuencias de las patologías que afectan a los grandes animales de Córdoba. El conocimiento geoespacial de las enfermedades es una herramienta esencial en el adecuado desempeño profesional.

Abstract This study examined the geographical distribution of bovine clinical casuistry in the outpatient service for large animals at the Universidad de Córdoba (Colombia). Only those complete clinical histories were considered for the study that presented a definitive diagnosis. The cases treated were classified according to the affected system, definitive diagnosis, type of treatment (medical or surgical), and geographical location in Córdoba. The study included 6356 clinical histories of cases presented between 2005 and 2016; 77.09% of them in the bovine species, and the most affected system was the hematopoietic system with 79.29% of casuistry, followed by the integumentary system (11.18%), and the digestive system (2.86%). 73% of the municipalities of the department were included, where a high prevalence of. It was established that the hematopoietic system was affected by endoglobular hemotropic parasites such as Babesia spp. and Anaplasma spp., while the integumentary system was mainly affected by infestations caused by Rhabditis spp. (41%), followed by dermatophytosis (19%) and papillomavirus infections (16%). It can be concluded that the outpatient service of the Medical-Surgical Clinic of Large Animals at the Universidad de Córdoba has a wide and varied casuistry, which is demonstrated in the frequency of pathologies that affect large animals in Córdoba. The geospatial knowledge of diseases is an essential tool for proper professional performance.

Resumo Neste estudo se realizou a distribuição geográfica da casuística clínica bovina do serviço ambulatório de Grandes Animais da Universidade de Córdoba (Colômbia). Foram consideradas dentro do estudo somente aquelas histórias clínicas completas que apresentaram um diagnóstico definitivo. Os casos atendidos foram classificados de acordo com o sistema afetado, o diagnóstico definitivo, o tipo de tratamento (médico ou cirúrgico) e a localização geográfica em Córdoba. Analisaram-se 6356 histórias clínicas de casos apresentados entre 2005 e 2016, das quais o 77,09 % foram na espécie bovina, e o sistema mais afetado foi o hematopoiético com 79,29 % da casuística, seguido do sistema tegumentário (11,18 %) e o sistema digestivo (2,86 %). Se atendeu o 73 % dos municípios do estado, donde se observou um alto predomínio de alterações no sistema hematopoiético (79,29 %), seguido pelo sistema tegumentário (11,18 %) e o sistema digestivo (2,86 %). Se estabeleceu que o sistema hematopoiético foi afetado por hemotrópicos endo globulares como Babesia spp. e Anaplasma spp., em quanto que o sistema tegumentário foi afetado principalmente por infestações com Rhabditis spp. (41 %), seguido de infecções por dermatofitose (19 %) e papiloma vírus (16 %). Se pode concluir que o serviço ambulatório da Clínica Médico-Cirúrgica de Grandes Animais da Universidade de Córdoba possui uma ampla e variada casuística, o que se demonstra nas frequências das patologias que afetam aos Grandes Animais de Córdoba. O conhecimento geoespacial das doenças é uma ferramenta essencial no adequado desempenho profissional.

Isolamento e caracterização de espécies de Pythium de ambientes aquáticos no Estado do Rio Grande do Sul e avaliação da patogenicidade em modelo experimental / Isolation and characterization of Pythium species from swampy areas in the Rio Grande do Sul, Brazil, and evaluation of pathogenicity in an experimental model

Foram coletadas 186 amostras de água de ambientes pantanosos em 13 municípios das regiões Sul, Central e Oeste do Rio Grande do Sul, Brasil, com o objetivo de isolar e caracterizar espécies de Pythium e avaliar a sua patogenicidade empregando coelhos como modelo experimental. Em 11,8% (n=22) das águas coletadas foram isoladas diferentes espécies de Pythium incluindo: P. insidiosum (n=1), P. catenulatum (n=3), P. pachycaule voucher (n=1), P. rhizo-oryzae (n=3), P. torulosum (n=4) e Pythium spp. (n=10). Zoósporos desses micro-organismos foram produzidos in vitro e inoculados por via subcutânea em coelhos, os quais foram avaliados durante 45 dias. Dentre os oomicetos testados, apenas P. insidiosum evidenciou patogenicidade, causando pitiose no modelo experimental, evidenciando que, em nossas condições, apenas esta espécie de Pythium é patógena para mamíferos.(AU)

One hundred and eighty-six water samples from swampy areas were collected in 13 municipalities of South, Central and West regions of Rio Grande do Sul, Brazil, in order to isolate and characterize Pythium species and assess their pathogenicity using rabbits as experimental model. Different Pythium species were isolated from 22 (11.8%) water samples, including P. insidiosum (n=1), P. catenulatum (n=3), P. pachycaule voucher (n=1), P. rhizo-oryzae (n=3), P. torulosum (n=4) e Pythium spp. (n=10). Zoospores of these microorganisms were produced in vitro and inoculated subcutaneously into rabbits, which were assessed over 45 days. Only P. insidiosum showed pathogenicity, causing pythiosis in the experimental model.(AU)

Pythiosis in dogs in the semiarid region of Northeast Brazil / Pitiose em cães na região semiárida do Nordeste, Brasil

We describe the epidemiological, clinical and pathological features of five cases of pythiosis in dogs in northeast semiarid. The disease occurred in dogs with age between one and three years, females and males of different breeds. The dogs were created in urban areas, but were occasionally taken to the countryside and then had contact with wetlands. The main clinical signs were weight loss, vomiting, diarrhea and tenesmus. Macroscopically there were intestinal wall thickening with irregular firm masses with yellowish granular areas interspersed with a whitish tissue, involving the duodenum, colon and rectum, extending to the lymph nodes and pancreas, and the vagina and liver. In the skin there were areas of alopecia and irregular ulcerations, some containing small cavitations with serosanguineous secretion. Microscopically there were pyogranulomatous inflammation and in one case also there were eosinophilic necrosis associated with negative images tubuliformes by hematoxylin and eosin, strongly stained by GMS and weakly by PAS. Hyphae were strongly marked by immunohistochemistry with polyclonal anti-Pythium insidiosum. The disease occurs sporadically in dogs in the semiarid Northeast, however should be included in the differential diagnosis of chronic diseases and proliferative aspect of the gastrointestinal system, and front skin lesions of difficult treatment.(AU)

Descrevem-se os aspectos epidemiológicos, clínicos e patológicos de cinco casos de pitiose em cães na região semiárida do Nordeste. A doença ocorreu em cães com idade entre um e três anos, fêmeas e machos, de diferentes raças. Os cães eram criados em zona urbana, mas ocasionalmente eram levados para zona rural e tinham contato com áreas alagadas. Em um caso não havia informações no histórico sobre acesso à zona rural. Os principais sinais clínicos observados foram perda de peso, vômito, diarreia e tenesmo. Macroscopicamente havia espessamento da parede do intestino com massas firmes, irregulares, com áreas granulares amareladas entremeadas por tecido esbranquiçado, no duodeno, cólon e reto, que se estendia ao pâncreas e linfonodos, além de fígado e vagina. Na pele havia área de alopecia e ulcerações irregulares, algumas contendo pequenas cavitações com secreção serosanguinolenta. Microscopicamente havia inflamação piogranulomatosa e em um caso também havia necrose eosinofílica associada a imagens negativas tubuliformes pela hematoxilina e eosina, fortemente impregnadas pelo GMS e fracamente coradas pelo PAS. As hifas foram fortemente imunomarcadas pela imuno-histoquímica com anticorpo policlonal anti-Pythium insidiosum. A doença ocorre esporadicamente em cães no semiárido nordestino, entretanto deve ser incluída no diagnostico diferencial das doenças em cães com evolução crônica e proliferativa que acometem o sistema gastrintestinal, e frente a lesões cutâneas de difícil tratamento.(AU)

Pythiosis cutaneous in horses treated with triamcinolone acetonide. Part 2. Histological and histochemical description / Pythiosis cutánea en equinos tratados con acetonida de triamcinolona. Parte 2. Descripción histológica e histoquímica

ABSTRACT Objective. The study aimed to evaluate the histomorphometry tissue recovery process of the skin granuloma of skin pythiosis in horses treated with triamcinolone acetonide. Materials and methods. We conducted a descriptive study, not probabilistic in convenience animals with cutaneous pythiosis. 24 horses were used with cutaneous pythiosis, a group of 12 animals was administered 50 mg of intramuscular injection of triamcinolone acetonide (TG) and the other group was not applied any treatment (CG). Are tissue biopsies performed for histological and histochemical evaluation and stained with hematoxylin eosin (HE), Gomori trichrome (GT), picrosirius red polarization (PR/P), Grocott methenamine silver (GMS) and periodic acid-Schiff (PAS). Results. It is noted that in TG inflammation was gradually decreasing, as evidenced in decreased fibrin layer leukocyte, PMN and phenomena Splendore Hoepli, also in increased angiogenesis, epiteliogénesis, and increasing the overall amount of fibroblasts and collagen fibers, anyway in the progressive replacement of collagen type III to type I collagen at the end of the process, and that the presence of intralesional pseudo-hyphae of Pythium insidiosum reduces it to the second week. Neither of the animals in the CG showed improvement in histological and histochemical characteristics of pythiosis and maintained equal to the first day throughout the study. Conclusions. The use of triamcinolone acetonide is a good therapeutic alternative for the treatment of granulomatous pythiosis wounds in horses with 100% clinical recovery and demonstrated with histological and histochemical findings.

RESUMEN Objetivo. El objetivo del estudio fue realizar una descripción histológica e histoquímica del proceso de cicatrización de la pythiosis cutánea en equinos tratados con acetonida de triamcinolona. Materiales y métodos. Se Realizó un estudio de tipo descriptivo, no probabilístico en animales de conveniencia con pythiosis cutánea. Fueron utilizados 24 equinos con pythiosis, siendo aplicado 50 mg de acetonida de triamcinolona vía intramuscular a un grupo de 12 animales (GT) y en otro grupo no fue aplicado tratamiento (GC). Se realizaron biopsias de tejidos para evaluación histológica e histoquímica, en las coloraciones de hematoxilina eosina (H&E), tricrómico de Gómori (TG), picrosirius red/polarization (PR/P) y plata metanamina de Grocott (GMS). Resultados. Se observó que en el GT el proceso inflamatorio fue disminuyendo progresivamente, evidenciado en la disminución de la capa fibrino leucocitaria, PMN y de los fenómenos de Splendore Hoepli, así como mayor angiogénesis, epiteliogénesis y aumento de la cantidad de fibroblastos y fibras colágenas generales, así mismo en el cambio progresivo de colágeno tipo III a colágeno tipo I al final del proceso, además de que la presencia de hifas intralesionales de Pythium insidiosum disminuyó a la segunda semana. Ninguno de los animales del GC presentó mejoría en las características histológicas e histoquímicas de la pythiosis y se mantuvieron iguales al primer día durante todo el estudio. Conclusiones. La acetonida de triamcinolona es una buena alternativa terapéutica en el tratamiento de las heridas granulomatosas por pythiosis en equinos con 100% de recuperación clínica y demostrada con las constataciones histológicas e histoquímicas.