Determination of the viability of Azospirillum syrups, multiplied in different culture media / Determinação da viabilidade de caldas de Azospirillum, multiplicadas em diferentes meios de cultura

The use of products of microbial origin, bacteria, for agriculture has grown exponentially in the world, and in Brazil this growth is significant. In recent years,several companies have settled in the country and started to develop different products in this segment. The producer has the possibility of multiplying bacteria on the farms themselves in a process called on farm, a homemade way of multiplying bacteria. The objective of this studywas to evaluate the efficiency of the mixtures produced through the on farmmethod of multiplication of microorganisms from inoculants based on Azospirillumwith different culture media. For this, three culture media were used: commercial product Multibacter®, yeast syrup plus sugar and yeast syrup, which were subsequently quantified. The syrups were produced in the onfarmsystem, the sterilization of the place of packaging and allocation of the bioreactors was carried out, the samples of the syrups were diluted for incubation and the preparation of the TSA(Trypticasein Soy Agar)also took place. It was possible to conclude that the non-sterile culture media do not present satisfactory results for the multiplication of on farmmicroorganisms with Azospirillumsyrups. Colony forming units are not parameters to indicate viability for non-sterile culture media.(AU)

O uso de produtos de origem microbiana, bactérias, para a agricultura tem crescido exponencialmente no mundo, e no Brasil este crescimento é significativo. Nos últimos anos várias empresas se instalaram no país e passaram a desenvolver diversos produtos neste segmento. O produtor, tem a possibilidade de multiplicar bactérias nas próprias fazendas em um processo chamado on farm,uma forma caseira de multiplicar bactérias. O objetivo deste estudo foi avaliar a eficiência das caldas produzidas através do método on farmde multiplicação de microrganismos de inoculantes a base de Azospirillumcom diferentes meios de culturas. Para isto foram utilizados três meios de cultura: produto comercial Multibacter®, calda de levedura mais açúcar e calda de levedura, os quais posteriormente foram quantificados. As caldas foram produzidas no sistema on farm, foi realizada a esterilização do local acondicionamento e alocação dos biorreatores, as amostras das caldas foram diluídas para incubação e também ocorreu o preparo do TSA (Trypticasein Soy Agar). Foi possível concluir que os meios de cultura não estéreis não apresentam resultados satisfatórios para a multiplicação de microrganismos on farmcom caldas de Azospirillum. Unidades formadoras de colônias não são parâmetros para indicar a viabilidade para meios de cultura não estéreis.(AU)

Amazonian soil fungi are efficient degraders of glyphosate herbicide; novel isolates of Penicillium, Aspergillus, and Trichoderma

Abstract Pesticide residues that contaminate the environment circulate within the hydrological cycle can accumulate within the food chain and cause problems to both environmental and human health. Microbes, however, are well known for their metabolic versatility and the ability to degrade chemically stable substances, including recalcitrant xenobiotics. The current study focused on bio-prospecting within Amazonian rainforest soils to find novel strains fungi capable of efficiently degrading the agriculturally and environmentally ubiquitous herbicide, glyphosate. Of 50 fungal strains isolated (using culture media supplemented with glyphosate as the sole carbon-substrate), the majority were Penicillium strains (60%) and the others were Aspergillus and Trichoderma strains (26 and 8%, respectively). All 50 fungal isolates could use glyphosate as a phosphorous source. Eight of these isolates grew better on glyphosate-supplemented media than on regular Czapek Dox medium. LC-MS revealed that glyphosate degradation by Penicillium 4A21 resulted in sarcosine and aminomethylphosphonic acid.

Resumo Resíduos de agrotóxicos que contaminam o meio ambiente circulam no ciclo hidrológico, podendo se acumular na cadeia alimentar e causar problemas tanto à saúde ambiental quanto humana. Por sua vez, microrganismos são bem conhecidos por sua versatilidade metabólica e capacidade de degradar substâncias quimicamente estáveis, incluindo xenobióticos recalcitrantes. O estudo atual se concentrou na bioprospecção nos solos da floresta amazônica para encontrar novas linhagens de fungos capazes de degradar com eficiência o herbicida onipresente na agricultura e no meio ambiente, o glifosato. Entre os 50 fungos isolados (usando meio de cultura suplementado com glifosato como única fonte de carbono), a maioria eram isolados do gênero Penicillium (60%) e os outros eram isolados de Aspergillus e Trichoderma (26 e 8%, respectivamente). Todos os 50 isolados de fungos foram capazes de usar glifosato como fonte de fósforo. Oito desses isolados cresceram melhor em meio suplementado com glifosato do que em meio Czapek Dox regular. LC-MS revelou que a degradação do glifosato por Penicillium 4A21 resultou nos metabólitos sarcosina e ácido aminometilfosfônico.

Canine ringworm caused by Trichophyton mentagrophytes - detection by SYBR-Green real-time PCR

Background: Dermatophytes, fungi of universal distribution, invade semi or fully keratinized structures, such as skin, fur/ hair and nails. The various species of dermatophytes are classified into three genera anamorphic: Microsporum, Trichophyton and Epidermophyton. The genus Epidermophyton includes only E. floccosum, that rarely affects animals. The main species responsible for the disease in dogs and cats are Microsporum canis, M. gypseum and Trichophyton mentagrophytes, which were characterized through conventional mycological methodology (microscopic examination with KOH and culture). Molecular methodologies, such as real-time PCR, can contribute to a rapid laboratory diagnosis, helping clinicians to initiate an early antifungal treatment. This case report describes a case of canine dermatophytosis due to Trichophyton mentagrophytes detected from a clinical sample by SYBR-Green real-time PCR. Case: A 8-year-old dog, rescued from the street, was referred to a private veterinary clinic in the city of Canoas, RS, Brazil, presenting generalized lymphadenomegaly, crusted lesions all over the body, generalized alopecia, signs of excoriation and epistaxis. Initially, were administered prednisone [1 mg/kg every 48 h, BID] and cephalexin [30 mg/kg, BID]. Weekly baths with benzoyl peroxide were also given. The therapy was not clinically successful. Wood's Lamp Test was negative. As a differential diagnosis, PCR for detection of Leishmania was negative. Complete blood count and serum biochemical assay were also performed. For mycological diagnosis, hair specimen was clarified and examined microscopically using 10% potassium hydroxide (KOH) for the visualization of chains of arthroconidia (ectothrix invasion of hair). The infected hair was plated onto MycoselTM Agar, incubated at 28°C for 15 days. Microscopy of hyphae/ conidia and macroscopic colony characteristics (colors and texture) were conducted for the differentiation of the species within the genus Microsporum and Trichophyton. In addition, real-time PCR was applied for direct analysis of the fungal DNA obtained from the hair sample. Microscopic examination was negative. The dermatophyte present in the hair sample was confirmed as Trichophyton mentagrophytes by culture and qPCR (melting-point analysis). The patient was treated with systemic itraconazole [10 mg/ kg SID - 90 days]. Twice-weekly application of 2.5 % miconazole and 2% chlorhexidine shampoo until complete cure. Discussion: Dermatophytosis is often listed as self-limiting infection; however, animal dermatophytosis can spread between pets, as well as a zoonotic transmission to humans. The literature on dermatophytosis indicates that Microsporum canis is the predominant etiological agent, followed by M. gypseum. Trichophyon mentagrophytes that appear in a lower percentage of isolation. The culture of hair, even with specific medium containing chloramphenicol and cyclohexamide, may present contaminating fungi, not related to dermatophytosis, which can inhibit or override the growth of dermatophytes. The use of real-time PCR provided a faster and specific diagnosis of dermatophytosis when compared to the conventional mycological methodology for detection and identification of T. mentagrophytes, which takes around 10 to 15 days for culture. It is possible to use this technique as an alternative diagnosis for dermatophytes associated to clinical hair samples of dogs.

Aspergilose em calopsita (Nymphicus hollandicus) / Aspergillosis in a cockatiel (Nymphicus hollandicus)

Background: Aspergillosis is a defiance in avian medicine, however between of Aspergillus species, 3 are the most frequently observed (A. flavus, A. fumigatus and A. niger). Psittaciformes present medical conditions related to the 3 species, however A. fumigatus is the most observed. The defiance of the diagnosis is related to fungus characteristics (they are saprophytic in healthy birds), the clinical signs and lesions, which are common to various respiratory diseases, and the difficulty to access specific diagnostic methods. Thus, we report a case of aspergillosis in a cockatiel, whose clinical diagnosis was confirmed by fungal culture of cloacal swab, and treatment with a fungistatic drug. Case: A female cockatiel, 2-month-old, not dewormed, presented diarrhea, sneezing and difficulty to gain weight, although eating well (extruded feed and supplementation with fruits and vegetables). At a clinical examination revealed body score (BCS) of 3 (scale 1 at 5), cloacal temperature (CT) of 40 ºC, dilated nostrils and edema in the left eye region associated to evident hyperemia. Due to it has lived with another animal, was inferred that was hypovitaminosis A, by food dispute and/ or dominance, or endoparasitosis, being prescribed vitamin A supplementation and deworming with ivermectin. The bird returned 15 days later, showing maintenance of weight loss, BCS 2, ruffled feathers and areas suggestive of apteriolysis in both forelimbs, respiratory difficulty and pulmonary and cranial air sacs rales, in addition presented wound in the pygostyle region and ventus, occasioned by attempted attack of a dog, and aggravated by self-mutilation. Due the worsening of the clinical condition, an oral cavity and cloacal swabs were collected to bacteriological and fungal culture. As treatment was prescribed: a natural tranquilizer, seeking to control the self-mutilation; doxycycline oral suspension, to treat a probable bacterial respiratory infection; and oral itraconazole suspension, manipulated and with a dose calculated by allometric scaling, being placed an elizabethan collar in order to prevent biting the pygostyle and ventus. With a positive result of the cloacal swab fungal culture for A. fumigatus, the doxycycline was discontinued. After the first 30 days of treatment with itraconazole, the patient returns to alert, weighing 95 g, BCS 3, with absence of respiratory signs and with signs of recent molting process. New swabs (oral and cloacal) were collected for repetition of bacterial and fungal cultures, being maintained the treatment with itraconazole. The results of both bacterial and fungal cultures were negative. At the end of the treatment with itraconazole, the patient returns demonstrating weight gain, EEC 3, CT of 42ºC, uniform feather coverage and absence of any respiratory sign, being released by the clinician. Discussion: A preponderant factor in the treatment of various respiratory diseases in birds is the diagnosis, since many of diseases present the same clinical signs. Thus, the use of fungal culture was essential to determine the origin of the signs. Still, even with an accurate diagnosis, the clinician often comes up against the difficulty of treatment, especially in cases of mycotic infections. The knowledge about the best drug, considering the pharmacology, as well as the availability of concentrations and dosages marketed, in addition to the long period of treatment, can difficult or make impossible the case resolution. In fact, this is confirmed by the literature, where the most of aspergillosis diagnosis are post mortem, resulting from the delay in seeking veterinary care or in the administration of ineffective drugs. Thus, with this report, we sought to present a diagnostic tool, sometimes neglected, and a compilation with the main drugs and their effectiveness in the treatment of aspergillosis in Psittaciformes.

Crescimentos de fungos produtores de micotoxinas em grãos de amendoim / Growths of mycotoxin-producing fungi on peanut grains

O amendoim (Arachis hypogaea L.) é uma oleaginosa com alto valor energético e nutricional, servindo de substrato ideal para o crescimento de fungos durante o processo de colheita, estocagem e processamento dos grãos. As micotoxinas são originadas a partir do metabolismo secundário de fungos toxigênicos; quando essas toxinas são ingeridas, podem causar efeitos tóxicos em humanos e animais. O objetivo deste trabalho foi avaliar a contaminação fúngica em amostras de grãos de amendoim disponíveis para o consumo humano. Foram analisadas amostras de amendoim torrado na vagem, amendoim cru (in natura) e amendoim torrado sem a vagem. A metodologia utilizada foi o método de plaqueamento direto em placas de Petri, contendo o meio cultura Sabouraud Dextrose Ágar (SDA). Porções de 40 grãos de amendoim de cada amostra foram utilizadas para o teste. Os resultados obtidos foram expressos em quantidade de grãos que apresentaram crescimento fúngico em porcentagem. Para identificação ao nível do género das colônias fúngicas isoladas, foi efetuada uma observação microscópica das suas estruturas morfológicas. Em todas as amostras estudadas ocorreram o crescimento de fungos potencialmente micotoxigênicos. Os grãos de amendoim cru (in natura) sem a vagem foram os mais contaminados por fungos como Aspergillus sp. (20%), Alternaria sp. (13,1%), Penicillium sp. (13,3%) e Fusarium sp. (23,3%) quando comparados com os grãos torrados. A presença desses fungos nas amostras de amendoim indica a possibilidade de desenvolvimento de micotoxinas potencialmente tóxicas para o humano. O processo de torrefação inibe o crescimento de determinadas espécies de fungos micotoxigênicos.(AU)

Peanut is an oilseed with high energy and nutritional value, serving as an ideal substrate for the growth of fungi during the process of harvesting, storing and processing the grains. Mycotoxins originate from the secondary metabolism of toxigenic fungi; when ingested, they can cause toxic effects in humans and animals. The objective of this work was to evaluate the fungal contamination in samples of peanut grains available for human consumption. The methodology used was the direct plating method in Petri dishes containing Sabouraud Dextrose Agar (SDA) culture. Portions of 40 peanut kernels from the samples were immersed in a 0.4% sodium hypochlorite solution for two minutes for surface disinfection, then rinsed once in distilled water. Incubation was carried out at room temperature for 7 days and the results were expressed as the amount of grains that showed fungal growth in percentage. To identify the colonies of isolated filamentous fungi at the genus level, a microscopic observation of their morphological structures was carried out, which consists of removing a sample from the edge of the colony and placing it in a drop of cotton blue (1g/L in lactic acid 88%), between slide and coverslip. Potentially toxigenic fungi were identified: Aspergillus sp., Rhizopus sp., Alternaria sp., Penicillium sp. and Fusarium sp. The presence of these fungi in peanut samples indicates the possibility of development of mycotoxins potentially toxic to humans. The roasting process may not be sufficient to destroy some mycotoxins.(AU)

Amazonian soil fungi are efficient degraders of glyphosate herbicide; novel isolates of Penicillium, Aspergillus, and Trichoderma / Os fungos do solo amazônico são degradadores eficientes do herbicida glifosato; novos isolados de Penicillium, Aspergillus e Trichoderma

Pesticide residues that contaminate the environment circulate within the hydrological cycle can accumulate within the food chain and cause problems to both environmental and human health. Microbes, however, are well known for their metabolic versatility and the ability to degrade chemically stable substances, including recalcitrant xenobiotics. The current study focused on bio-prospecting within Amazonian rainforest soils to find novel strains fungi capable of efficiently degrading the agriculturally and environmentally ubiquitous herbicide, glyphosate. Of 50 fungal strains isolated (using culture media supplemented with glyphosate as the sole carbon-substrate), the majority were Penicillium strains (60%) and the others were Aspergillus and Trichoderma strains (26 and 8%, respectively). All 50 fungal isolates could use glyphosate as a phosphorous source. Eight of these isolates grew better on glyphosate-supplemented media than on regular Czapek Dox medium. LC-MS revealed that glyphosate degradation by Penicillium 4A21 resulted in sarcosine and aminomethylphosphonic acid.

Resíduos de agrotóxicos que contaminam o meio ambiente circulam no ciclo hidrológico, podendo se acumular na cadeia alimentar e causar problemas tanto à saúde ambiental quanto humana. Por sua vez, microrganismos são bem conhecidos por sua versatilidade metabólica e capacidade de degradar substâncias quimicamente estáveis, incluindo xenobióticos recalcitrantes. O estudo atual se concentrou na bioprospecção nos solos da floresta amazônica para encontrar novas linhagens de fungos capazes de degradar com eficiência o herbicida onipresente na agricultura e no meio ambiente, o glifosato. Entre os 50 fungos isolados (usando meio de cultura suplementado com glifosato como única fonte de carbono), a maioria eram isolados do gênero Penicillium (60%) e os outros eram isolados de Aspergillus e Trichoderma (26 e 8%, respectivamente). Todos os 50 isolados de fungos foram capazes de usar glifosato como fonte de fósforo. Oito desses isolados cresceram melhor em meio suplementado com glifosato do que em meio Czapek Dox regular. LC-MS revelou que a degradação do glifosato por Penicillium 4A21 resultou nos metabólitos sarcosina e ácido aminometilfosfônico.

Amazonian soil fungi are efficient degraders of glyphosate herbicide; novel isolates of Penicillium, Aspergillus, and Trichoderma / Os fungos do solo amazônico são degradadores eficientes do herbicida glifosato; novos isolados de Penicillium, Aspergillus e Trichoderma

Pesticide residues that contaminate the environment circulate within the hydrological cycle can accumulate within the food chain and cause problems to both environmental and human health. Microbes, however, are well known for their metabolic versatility and the ability to degrade chemically stable substances, including recalcitrant xenobiotics. The current study focused on bio-prospecting within Amazonian rainforest soils to find novel strains fungi capable of efficiently degrading the agriculturally and environmentally ubiquitous herbicide, glyphosate. Of 50 fungal strains isolated (using culture media supplemented with glyphosate as the sole carbon-substrate), the majority were Penicillium strains (60%) and the others were Aspergillus and Trichoderma strains (26 and 8%, respectively). All 50 fungal isolates could use glyphosate as a phosphorous source. Eight of these isolates grew better on glyphosate-supplemented media than on regular Czapek Dox medium. LC-MS revealed that glyphosate degradation by Penicillium 4A21 resulted in sarcosine and aminomethylphosphonic acid.(AU)

Resíduos de agrotóxicos que contaminam o meio ambiente circulam no ciclo hidrológico, podendo se acumular na cadeia alimentar e causar problemas tanto à saúde ambiental quanto humana. Por sua vez, microrganismos são bem conhecidos por sua versatilidade metabólica e capacidade de degradar substâncias quimicamente estáveis, incluindo xenobióticos recalcitrantes. O estudo atual se concentrou na bioprospecção nos solos da floresta amazônica para encontrar novas linhagens de fungos capazes de degradar com eficiência o herbicida onipresente na agricultura e no meio ambiente, o glifosato. Entre os 50 fungos isolados (usando meio de cultura suplementado com glifosato como única fonte de carbono), a maioria eram isolados do gênero Penicillium (60%) e os outros eram isolados de Aspergillus e Trichoderma (26 e 8%, respectivamente). Todos os 50 isolados de fungos foram capazes de usar glifosato como fonte de fósforo. Oito desses isolados cresceram melhor em meio suplementado com glifosato do que em meio Czapek Dox regular. LC-MS revelou que a degradação do glifosato por Penicillium 4A21 resultou nos metabólitos sarcosina e ácido aminometilfosfônico.(AU)

Criptococose respiratória e cutânea em gato de vida livre da cidade de Sobral/CE / Respiratory and cutaneous Cryptococcosis in a free-range cat in the city of Sobral/CE

A criptococose é uma doença infecciosa sistêmica de origem fúngica, causada pelas espécies: Cryptococcus neoformans e Cryptococcus gattii. É considerada uma micose oportunista capaz de infectar mamíferos domésticos, animais silvestres e seres humanos, sendo classificada como uma doença zoonótica. Esse patógeno é encontrado principalmente em ambientes contaminados por fezes de pombos (Columba livia), que atuam como importantes fontes de infecção do fungo. De acordo com sua disseminação para os tecidos do organismo, a doença pode causar diferentes síndromes tanto em seres humanos como em animais síndrome respiratória, síndrome neurológica, síndrome ocular e síndrome cutânea. O diagnóstico pode ser realizado através da pesquisa de antígeno polissacarídeo circulante no soro ou líquor, por meio da prova de látex. Testes imunoenzimáticos (ELISA) também podem ser realizados para detecção de antígenos dessa levedura. Exame citológico, histopatológico e cultura fúngica para a identificação do agente tornam o diagnóstico da criptococose mais fácil. O tratamento é baseado no uso de antifúngicos e sua escolha é realizada através da avaliação dos sinais clínicos observados. Diante do exposto, o objetivo deste trabalho é relatar um caso de criptococose respiratória e cutânea em felino doméstico de vida livre da cidade de Sobral/CE.

Cryptococcosis is a systemic infectious disease of fungal origin caused by the species: Cryptococcus neoformans and Cryptococcus gattii. It is considered an opportunistic mycosis capable of infecting domestic mammals, wild animals, and humans, being classified as a zoonotic disease. This pathogen is mainly found mainly in environments contaminated by pigeon (Columba livia) feces, which act as important sources of fungal infection. According to its spread to the body's tissues, the disease can cause different syndromes in both humans and animals: respiratory syndrome, neurological syndrome, ocular syndrome, and cutaneous syndrome. The diagnosis can be made through the investigation of circulating polysaccharide antigen in serum or cerebrospinal fluid using the latex test. Immunoenzymatic tests (ELISA) can also be performed to detect yeast antigens. Cytological examination, histopathological examination, and fungal culture to identify the agent make the diagnosis of cryptococcosis easier. The treatment is based on the use of antifungals, and its choice is made through the evaluation of the observed clinical signs. In this context, this work aims to report a case of respiratory and cutaneous cryptococcosis in a free-range domestic cat in the city of Sobral/CE.

Optimization of oxalic acid production by fungi for biotechnological solubilization of rock phosphate

This study describes a biotechnological strategy for producing and applying oxalic acid to solubilize phosphorus (P) from rock phosphate (RP). We evaluated six fungal species (Aspergillus niger FS1, Penicillium islandicum FS41, Pleurotus ostreatus PO1, Rhizoctonia solani Rhiz555, Sclerotium rolfsii Sr25, and Sclerotinia sclerotiorum Scl134) and three culture media (potato dextrose broth, Tsao and Strasser media) to maximize oxalic acid production. Among the fungal isolates tested and culture media, S. rolfsii Sr25 and Tsao medium showed efficient oxalic acid production. Tsao medium was optimized following a response surface methodology after initial screening of factors affecting RP solubilization. The optimized concentrations were 1 g L-1 NaNO3, 100 g L-1 glucose, 2 g L-1 KH2PO4, 4.5 g L-1 yeast extract, and 25 mg L-1 MgSO4·7H2O used for 20 days of incubation. Under these conditions, 71 mmol L-1 oxalic acid was obtained, representing a three-fold increase over production under non-optimized conditions (20 mmol L-1). Under optimized conditions, oxalic acid produced by S. rolfsii Sr25 reacted with low-solubility RP and solubilized 100 % of the P contained in ore. Thus, using S. rolfsii Sr25 to produce oxalic acid seems a promising biotechnological alternative for P solubilization from RP.(AU)

Sporothrix brasiliensis infecting cats in northeastern Brazil: New emerging areas in Paraíba state / Sporothrix brasiliensis infectando felinos no nordeste do Brasil: novas áreas emergentes no estado da Paraíba

ABSTRACT: Sporotrichosis is a subcutaneous mycosis caused by fungus of the Sporothrix complex, and in Brazil the main species reported is Sporothrix brasiliensis, of which the diseased cat is the transmitter. Although, its occurrence has increased in the state of Paraíba, Brazil, since 2016, data on the disease in this state are limited. Therefore, this research aimed to identify molecularly isolates of Sporothrix spp. from domestic cats from cities in Paraíba, and in this way to expand the understanding of the disease in the state. Thirty-nine samples were analyzed, obtained from skin lesions of domestic felines, from the following cities in Paraíba: João Pessoa, Pilões, Patos, Areia, Bananeiras and Guarabira. Cytological analysis was performed to screen the samples, followed by fungal culture, and the molecular characterization of the isolates was performed, using the species-specific Polymerase Chain Reaction (PCR) or partial sequencing of the calmodulin gene. All isolates were identified as S. brasiliensis. The sequencing showed 100% similarity to the S. brasiliensis CBS 120339 strain. In view of this, it is concluded that in the study areas the species involved in cases of feline sporotrichosis is S. brasiliensis, its presence in Paraíba demonstrated the spread of the agent in regions distant from the epicenters in Brazil, alerting to the possible occurrence of zoonotic outbreaks similar to those found in the South and Southeast regions of the country. In addition, it highlights the emerging role of felines in the transmission of sporotrichosis in new endemic areas of Brazil.

RESUMO: A esporotricose é uma micose subcutânea causada por fungos do complexo Sporothrix, e no Brasil a principal espécie relatada é Sporothrix brasiliensis da qual o transmissor é o gato doente. Embora sua ocorrência tenha aumentado no estado da Paraíba, Brasil, desde 2016, os dados sobre a doença neste estado são limitados. Diante disso, esta pesquisa teve por objetivo identificar, molecularmente, isolados de Sporothrix spp. procedentes de felinos domésticos de cidades da Paraíba, e dessa maneira expandir a compreensão da enfermidade no estado. Foram analisadas 39 amostras, obtidas de lesões cutâneas de felinos domésticos, oriundos das seguintes cidades paraibanas: João Pessoa, Pilões, Patos, Areia, Bananeiras e Guarabira. Realizou-se análise citológica, para triagem das amostras, a seguir cultura fúngica, e posteriormente a caracterização molecular dos isolados, utilizando a Reação em Cadeia da Polimerase (PCR), espécie-específica ou sequenciamento parcial do gene calmodulina. Todos os isolados foram identificados molecularmente como S. brasiliensis. O sequenciamento demonstrou 100% de similaridade com a cepa S. brasiliensis CBS 120339. Contudo, conclui-se que nas áreas do estudo a espécie envolvida em casos de esporotricose felina é S. brasiliensis, sua presença na Paraíba demonstra a disseminação do agente em regiões distantes dos epicentros no Brasil, alertando para a possível ocorrência de surtos zoonóticos, semelhantes aos encontrados nas regiões Sul e Sudeste do país. Além disso, destaca o papel emergente dos felinos na transmissão da esporotricose em novas áreas endêmicas do Brasil.

Amazonian soil fungi are efficient degraders of glyphosate herbicide; novel isolates of Penicillium, Aspergillus, and Trichoderma / Os fungos do solo amazônico são degradadores eficientes do herbicida glifosato; novos isolados de Penicillium, Aspergillus e Trichoderma

Abstract Pesticide residues that contaminate the environment circulate within the hydrological cycle can accumulate within the food chain and cause problems to both environmental and human health. Microbes, however, are well known for their metabolic versatility and the ability to degrade chemically stable substances, including recalcitrant xenobiotics. The current study focused on bio-prospecting within Amazonian rainforest soils to find novel strains fungi capable of efficiently degrading the agriculturally and environmentally ubiquitous herbicide, glyphosate. Of 50 fungal strains isolated (using culture media supplemented with glyphosate as the sole carbon-substrate), the majority were Penicillium strains (60%) and the others were Aspergillus and Trichoderma strains (26 and 8%, respectively). All 50 fungal isolates could use glyphosate as a phosphorous source. Eight of these isolates grew better on glyphosate-supplemented media than on regular Czapek Dox medium. LC-MS revealed that glyphosate degradation by Penicillium 4A21 resulted in sarcosine and aminomethylphosphonic acid.

Resumo Resíduos de agrotóxicos que contaminam o meio ambiente circulam no ciclo hidrológico, podendo se acumular na cadeia alimentar e causar problemas tanto à saúde ambiental quanto humana. Por sua vez, microrganismos são bem conhecidos por sua versatilidade metabólica e capacidade de degradar substâncias quimicamente estáveis, incluindo xenobióticos recalcitrantes. O estudo atual se concentrou na bioprospecção nos solos da floresta amazônica para encontrar novas linhagens de fungos capazes de degradar com eficiência o herbicida onipresente na agricultura e no meio ambiente, o glifosato. Entre os 50 fungos isolados (usando meio de cultura suplementado com glifosato como única fonte de carbono), a maioria eram isolados do gênero Penicillium (60%) e os outros eram isolados de Aspergillus e Trichoderma (26 e 8%, respectivamente). Todos os 50 isolados de fungos foram capazes de usar glifosato como fonte de fósforo. Oito desses isolados cresceram melhor em meio suplementado com glifosato do que em meio Czapek Dox regular. LC-MS revelou que a degradação do glifosato por Penicillium 4A21 resultou nos metabólitos sarcosina e ácido aminometilfosfônico.

Meningoencephalitis of unknown origin in dogs

Background: Meningoencephalitis of unknown origin (MUO) is a critical cause of neurological disorders in dogs, mainly affecting small young individuals. Its symptomatology is varied and depends on the affected neuroanatomic region. The ante mortem diagnosis of this condition is uncertain, being achieved by discarding other conditions and often occurring definitively only by performing a necropsy. Thus, this study aims to report 2 cases of meningoencephalitis, one necrotizing and the other granulomatous in dogs. Cases: Case 1. A 3-year-old, Shih Tzu bitch with a body weight of 4 kg, showing proprioceptive ataxia, behavior of walking in circles, and evolving rapidly to non-ambulatory paresis, was treated. The neurological examination showed a posture of decerebrate stiffness and absence of withdrawal reflex and proprioception, suggesting brainstem injury. Laboratory tests showed mild neutrophilia and lymphopenia, while the rapid test for distemper was non-reactive. The cerebrospinal fluid (CSF) analysis showed lymphocytic pleocytosis, and the PCR tests of the CSF, blood, and urine for the detection of infectious diseases were negative, as well as the culture. With no improvement in clinical condition and exams showing a progressive degenerative condition unresponsive to available treatments, the tutor opted for euthanasia of the patient. The subsequently requested necropsy confirmed the diagnosis of granulomatous meningoencephalitis. Case 2. This case refers to a 1-year-and-5-month-old male Maltese breed weighing 4.8 kg. This animal presented walking in circles behavior and loss of vision for a week, with signs worsening rapidly. In the neurological evaluation, the patient presented sensitivity in the middle ear, difficulty opening the mouth, hearing deficit in the right ear, blindness in the right eye, a proprioceptive deficit in the right anterior limb, and head pressing. Laboratory tests showed nonregenerative anemia and mild lymphopenia. After 1 day of hospitalization, the patient showed worsening clinical condition, with obstruction, absence of facial and auricular sensitivity, and nasal stimulus. In addition, onset of generalized seizures was observed; therefore, CSF was analyzed, which did not present significant alterations except for detecting reactive lymphocytes. The bacteriological culture of CSF resulted in no bacterial growth. In addition, the same neurological PCR panel performed for the previous patient was negative. After 5 days of intensive care, the patient presented a cardiorespiratory arrest and died. The subsequently requested necropsy confirmed the diagnosis of necrotizing meningoencephalitis. Discussion: The 2 reported cases confirm that MUO should be considered during the differential diagnoses of patients with neurological alterations. It is known that small-breed dogs are predisposed to these diseases. Laboratory tests and medical imaging are crucial for clinical guidance, helping to discard other neurological pathologies, especially those due to bacterial, fungal, and/or viral agents. However, definitive diagnosis of MUO can only be performed through necropsy and histopathological analysis. For the reported cases, CSF analysis, neurological PCR panel for detecting possible infectious agents, and bacterial culture were essential to rule out other possible causes of meningoencephalitis. Unfortunately, MUO includes progressive neurological disorders causing the patient's death.

Sporothrix brasiliensis infecting cats in northeastern Brazil: New emerging areas in Paraíba state / Sporothrix brasiliensis infectando felinos no nordeste do Brasil: novas áreas emergentes no estado da Paraíba

Sporotrichosis is a subcutaneous mycosis caused by fungus of the Sporothrix complex, and in Brazil the main species reported is Sporothrix brasiliensis, of which the diseased cat is the transmitter. Although, its occurrence has increased in the state of Paraíba, Brazil, since 2016, data on the disease in this state are limited. Therefore, this research aimed to identify molecularly isolates of Sporothrix spp. from domestic cats from cities in Paraíba, and in this way to expand the understanding of the disease in the state. Thirty-nine samples were analyzed, obtained from skin lesions of domestic felines, from the following cities in Paraíba: João Pessoa, Pilões, Patos, Areia, Bananeiras and Guarabira. Cytological analysis was performed to screen the samples, followed by fungal culture, and the molecular characterization of the isolates was performed, using the species-specific Polymerase Chain Reaction (PCR) or partial sequencing of the calmodulin gene. All isolates were identified as S. brasiliensis. The sequencing showed 100% similarity to the S. brasiliensis CBS 120339 strain. In view of this, it is concluded that in the study areas the species involved in cases of feline sporotrichosis is S. brasiliensis, its presence in Paraíba demonstrated the spread of the agent in regions distant from the epicenters in Brazil, alerting to the possible occurrence of zoonotic outbreaks similar to those found in the South and Southeast regions of the country. In addition, it highlights the emerging role of felines in the transmission of sporotrichosis in new endemic areas of Brazil.

A esporotricose é uma micose subcutânea causada por fungos do complexo Sporothrix, e no Brasil a principal espécie relatada é Sporothrix brasiliensis da qual o transmissor é o gato doente. Embora sua ocorrência tenha aumentado no estado da Paraíba, Brasil, desde 2016, os dados sobre a doença neste estado são limitados. Diante disso, esta pesquisa teve por objetivo identificar, molecularmente, isolados de Sporothrix spp. procedentes de felinos domésticos de cidades da Paraíba, e dessa maneira expandir a compreensão da enfermidade no estado. Foram analisadas 39 amostras, obtidas de lesões cutâneas de felinos domésticos, oriundos das seguintes cidades paraibanas: João Pessoa, Pilões, Patos, Areia, Bananeiras e Guarabira. Realizou-se análise citológica, para triagem das amostras, a seguir cultura fúngica, e posteriormente a caracterização molecular dos isolados, utilizando a Reação em Cadeia da Polimerase (PCR), espécie-específica ou sequenciamento parcial do gene calmodulina. Todos os isolados foram identificados molecularmente como S. brasiliensis. O sequenciamento demonstrou 100% de similaridade com a cepa S. brasiliensis CBS 120339. Contudo, conclui-se que nas áreas do estudo a espécie envolvida em casos de esporotricose felina é S. brasiliensis, sua presença na Paraíba demonstra a disseminação do agente em regiões distantes dos epicentros no Brasil, alertando para a possível ocorrência de surtos zoonóticos, semelhantes aos encontrados nas regiões Sul e Sudeste do país. Além disso, destaca o papel emergente dos felinos na transmissão da esporotricose em novas áreas endêmicas do Brasil.

Coriandrum sativum L. essential oil obtained from organic culture shows antifungal activity against planktonic and multibiofilm Candida / Óleo essencial de Coriandrum sativum L. obtido de cultura orgânica apresenta atividade antifúngica contra Candida planctônica e multibiofilme

This study aimed to analyze the phytochemical profile of essential oil obtained from the leaves of Coriandrum sativum L., and its antifungal activity against Candida spp. The research consisted of an in vitro study including collecting the vegetable product, analysis of its macronutrients, extraction, and chemical analysis of the essential oil, and assaying antifungal activity through minimum inhibitory concentration (MIC) and minimum fungicidal concentration (MFC), with growth inhibition kinetics, and the product's effects on multi-species Candida biofilm. Nitrogen (47.08 g Kg-1), phosphorus (5.3 g Kg-1) and potassium (50.46 g Kg-1) levels were within the normal range. The major constituents were octanal, decanal, dec-(2E)-enal, and dodecanal. The MIC and MFC of the product evaluated against 11 tested Candida strains ranged from 31.25 to 250 µg/mL. There was inhibition of fungal growth during 24 hours of exposure at the 3 concentrations tested (250, 125, and 62.5 µg/mL). The concentration of 80 mg/mL promoted the greatest reduction in multispecies biofilm (70% reduction in biofilm). Coriandrum sativum L. essential oil extract is principally constituted of alcohols and aldehydes and presents fungicidal activity against Candida spp. in its in planktonic and biofilm forms.

Este trabalho teve como objetivo analisar o perfil fitoquímico do óleo essencial obtido das folhas de Coriandrum sativum L., e sua atividade antifúngica contra Candida spp. A pesquisa consistiu em um estudo in vitro incluindo a coleta do produto vegetal, análise de seus macronutrientes, extração e análise química do óleo essencial e ensaio da atividade antifúngica por meio da concentração inibitória mínima (CIM) e concentração fungicida mínima (MFC), com crescimento cinética de inibição e os efeitos do produto no biofilme de Candida multi-espécies. Os níveis de nitrogênio (47,08 g Kg-1), fósforo (5,3 g Kg-1) e potássio (50,46 g Kg-1) estavam dentro da normalidade. Os principais constituintes foram octanal, decanal, dec-(2E)-enal e dodecanal. A CIM e CFM do produto avaliado contra 11 cepas de Candida testadas variaram de 31,25 a 250 µg/mL. Houve inibição do crescimento fúngico durante 24 horas de exposição nas 3 concentrações testadas (250, 125 e 62,5 µg/mL). A concentração de 80 mg/mL promoveu a maior redução no biofilme multiespécies (redução de 70% no biofilme). O extrato do óleo essencial de Coriandrum sativum L. é constituído principalmente por álcoois e aldeídos e apresenta atividade fungicida contra Candida spp. em suas formas planctônicas e biofilme.

Histoplasmose felina / Feline histoplasmosis

A histoplasmose é uma doença zoonótica, causada por um fungo oportunista, chamado Histoplasma capsulatum. O fungo é endêmico de regiões subtropicais e temperadas, sendo sua presença relatada principalmente nas Américas, na Índia e no Sudoeste Asiático.Acredita-se que os casos de histoplasmose em gatos ainda sejam subestimados. No Brasil, a maioria dos casos relatados foram provenientes do estado do Ceará. A histoplasmose é considerada a segunda causa mais comum de afecção micótica sistêmica em gatos. Desta forma, o objetivo do presente trabalho foi relatar a ocorrência de um caso com resolução satisfatória em Fortaleza/Ceará. O animal apresentou histórico de espirros crônicos, secreção nasal e um aumento de volume na região nasal, além de um nódulo ulcerado no flanco direito. Foram solicitadas citologia e cultura da lesão, onde foram evidenciadas leveduras sugestivas de Histoplasma capsulatum. O tratamento escolhido foi itraconazol na dose de 100mg/gato, sendo administrada por via oral uma cápsula ao dia. Após 30 dias, o animal retornou ao consultório, apresentando uma melhora considerável, sem aumento de plano nasal, processo obstrutivo nasal e lesões de pele.

Histoplasmosis is a zoonotic disease caused by an opportunistic fungus called Histoplasma capsulatum. The fungus is endemic to subtropical and temperate regions, with its presence being reported mainly in the Americas, India, and Southeast Asia. It is believed that the cases of histoplasmosis in cats are still underestimated. In Brazil, most reported cases were from the state of Ceará. Histoplasmosis is considered the second most common cause of systemic mycotic disease in cats. Thus, the present study aimed to report the occurrence of a case with satisfactory resolution in Fortaleza/Ceará. The animal showed a history of chronic sneezing, nasal secretion, and an increase in volume in the nasal region, in addition to an ulcerated nodule on the right flank. Cytology and culture of the lesion were requested, where yeasts suggestive of Histoplasma capsulatum were evidenced. The chosen treatment was itraconazole at a dose of 100mg/cat, being administered orally one capsule a day. After 30 days, the animal returned to the clinic showing considerable improvement, without an increase in the nasal plane, nasal obstructive process, and skin lesions.

Cutaneous candidiasis caused by antifungal-resistant Candida sp. strain in canine individual / Candidíase cutânea causada por cepa de Candida sp. resistente a antifúngicos em indivíduo canino

The aim of the current report is to describe a cutaneous candidiasis case affecting a canine individual treated at the University Veterinary Hospital of State University of Maranhão (UEMA), in São Luís City. The patient had three-month history of skin diseases; it had been previously subjected to several treatments based on antibiotics, corticosteroids and antifungal drugs that have failed to show clinical improvements. Dermatological assessment has indicated generalized moist dermatitis, intense skin desquamation, alopecia, pruritus and meliceric crusts along the animal's body, mainly in its dorsal region. Complementary tests, such as skin cytology and microscopy, trichogram, qualitative PCR and serology for canine visceral leishmaniasis, as well as fungal culture and antifungigram were requested based on this scenario. Serology recorded inconclusive results for leishmaniasis, whereas PCR recorded negative results in the presence of the agent's DNA. Cytology, microscopy and trichogram results have evidenced fungal infection in the assessed samples. Moreover, mycological culture and antifungigram resulted in the growth of Candida sp. specimens capable of resisting antifungal agents such as amphotericin B, fluconazole, itraconazole and nystatin. The therapy adopted after candidiasis diagnosis confirmation comprised oral doses of manipulated ketoconazole, in combination to topical therapy with shampoo based on moisturizing formulas associated with Miconazole and Chlorhexidine (at 2%), for four weeks. After 30 days, when the adopted therapy was over, the aforementioned animal presented remission of the previously observed lesions and fully improved condition.

The aim of the current report is to describe a cutaneous candidiasis case affecting a canine individual treated at the University Veterinary Hospital of State University of Maranhão (UEMA), in São Luís City. The patient had three-month history of skin diseases; it had been previously subjected to several treatments based on antibiotics, corticosteroids and antifungal drugs that have failed to show clinical improvements. Dermatological assessment has indicated generalized moist dermatitis, intense skin desquamation, alopecia, pruritus and meliceric crusts along the animal's body, mainly in its dorsal region. Complementary tests, such as skin cytology and microscopy, trichogram, qualitative PCR and serology for canine visceral leishmaniasis, as well as fungal culture and antifungigram were requested based on this scenario. Serology recorded inconclusive results for leishmaniasis, whereas PCR recorded negative results in the presence of the agent's DNA. Cytology, microscopy and trichogram results have evidenced fungal infection in the assessed samples. Moreover, mycological culture and antifungigram resulted in the growth of Candida sp. specimens capable of resisting antifungal agents such as amphotericin B, fluconazole, itraconazole and nystatin. The therapy adopted after candidiasis diagnosis confirmation comprised oral doses of manipulated ketoconazole, in combination to topical therapy with shampoo based on moisturizing formulas associated with Miconazole and Chlorhexidine (at 2%), for four weeks. After 30 days, when the adopted therapy was over, the aforementioned animal presented remission of the previously observed lesions and fully improved condition.

Refractory feline sporotrichosis: a comparative analysis on the clinical, histopathological, and cytopathological aspects

ABSTRACT: Sporotrichosis is a chronic fungal infection caused by Sporothrix species. The occurrence of cases that are resistant to long-term treatment, especially in the nasal planum of cats, emphasizes the importance of studying its pathogenesis. The purpose of this study was to analyze and compare the inflammatory process of cutaneous lesions of feline refractory sporotrichosis to clinical aspects through cytopathological and histopathological examination. Moreover, the study included 13 cats with cutaneous lesions that had been resistant to itraconazole treatment for more than a year. Cutaneous lesions samples were collected for cytopathological, histopathological, and fungal culture analyses. Tissue fragments were processed and stained with hematoxylin-eosin (HE) and Grocott methenamine silver (GMS). Further, two clinical presentations had the highest occurrence: the localized cutaneous form in animals with good general condition and stable disease (n=9, 69.2%) and the disseminated cutaneous form in cats with poor general condition (n=4, 30.8%). In cats with refractory sporotrichosis, the nasal planum (84.6%) was the most common location of lesions. In the cytopathological study, cats with fewer than two lesions and in good general condition (n=9, 69.2%) showed absence or mild yeast intensity (up to 5 yeasts per field), lower intensity of macrophages and neutrophils, and higher intensity of epithelioid cells, lymphocytes, plasma cells, and eosinophils. On the other hand, (n=4, 30.8%) of the cats with disseminated sporotrichosis and a poor general condition had a marked intensity of yeasts, which were mostly phagocytosed by an increased number of macrophages and neutrophils. Of those animals with good general condition, the majority (n=6, 66.7%) had higher eosinophil intensity. In histopathology, malformed suppurative granuloma was the predominant type (n=9, 69.2%) in feline sporotrichosis lesions, followed by well-formed granulomas (n=4, 30.8%). Malformed granulomas showed mild to moderate fungal intensity (55.6%) in animals with good general condition and localized lesions while marked fungal intensity (44.4%) in cats with the disseminated form of the disease and poor general condition. Well-formed granulomas (n=4, 30.7%) had mild to moderate intensity of fungal load, and 75% of the animals with this type of granuloma had only one lesion and were in good general condition. Long-term itraconazole treatment in these cats with refractory sporotrichosis can keep the infection under control and localized lesions stable; however, fungus reactivation can occur, resulting in an exuberant and inefficient immune response.

RESUMO: A esporotricose é uma infecção fúngica crônica causada por espécies do gênero Sporothrix. A ocorrência de casos refratários ao tratamento de longo prazo, especialmente na região nasal de gatos, alerta para a importância do estudo de sua patogênese. Este estudo teve como objetivo analisar o processo inflamatório das lesões cutâneas de esporotricose felina refratária ao tratamento, por meio de exames citopatológico e histopalógico, e comparar com aspectos clínicos. Treze gatos com lesões cutâneas refratárias ao tratamento com Itraconazol por mais de um ano foram incluídos no estudo. Amostras de lesões cutâneas foram coletadas para análises citopatológicas, histopatológicas e cultura fúngica. Fragmentos de tecidos foram processados e corados pela hematoxilina-eosina (HE) e Prata Metenamina de Grocott (GMS). Duas apresentações clínicas tiveram maior ocorrência: a forma localizada cutânea em animais com bom estado geral e doença estável (n=9, 69,2%); e a forma disseminada cutânea em gatos com estado geral ruim (n=4, 30,8%). A região nasal (84,6%) foi a localização mais frequente das lesões nos gatos com esporotricose refratária ao tratamento. No estudo citopatológico, felinos com menos de duas lesões, e em bom estado geral, (n=9, 69,2%) revelaram ausência ou leve intensidade de leveduras (até 5 leveduras por campo), menor intensidade de macrófagos e neutrófilos, e maior intensidade de células epitelióides, linfócitos, plasmócitos e eosinófilos. Enquanto, (n=4, 30,8%) dos felinos que apresentavam a forma disseminada da esporotricose associada a um estado geral ruim, revelaram acentuada intensidade de leveduras em sua maioria fagocitadas por acentuado número de macrófagos e neutrófilos. Dos animais com bom estado geral, a maioria (n=6, 66,7%) apresentava maior intensidade de eosinófilos. Na histopatologia, o granuloma supurativo mal formado foi o tipo predominante (n=9, 69,2%) nas lesões de esporotricose felina, e (n=4, 30,8%) foram de granulomas bem formados. Granulomas mal formados apresentaram leve a moderada intensidade fúngica (55,6%) nos animais com bom estado geral e lesões localizadas, e acentuada intensidade fúngica (44,4%) nos gatos com a forma disseminada da doença e estado geral ruim. Granulomas bem formados (n=4, 30,7%) apresentaram leve a moderada intensidade de carga fúngica, sendo 75% dos animais com esse tipo de granuloma com apenas uma lesão e bom estadogeral. O tratamento de longo prazo com itraconazol, nesses gatos com esporotricose refratária, pode manter a infecção controlada e lesões localizadas estáveis de forma temporária, contudo a reativação do fungo pode ocorrer levando a uma resposta imunológica exuberante e ineficiente.

Bacillus subtilis Bs10 as an efficient inoculant for growth promotion in soybean plants / Bacillus subtilis Bs10 como um inoculante eficiente para a promoção do crescimento de plantas de soja

Currently, food with low levels of pesticides, sustainable agricultural practices and increased crop productivity have gained prominence worldwide. Rhizobacterial inoculants are an alternative to manage large crops that favors sustainable plant growth. Thereby, this study aimed to assess the effect of increasing doses of inoculant based on the rhizobacterium Bacillus subtilis Bs10 compared to the commercial product based on B. subtilis on the development of soybean culture. Two experiments were carried out with two soybean cultivars, M8210 IPRO and M8615 IPRO, assessed at the vegetative and reproductive stages. Doses of B. subtilis Bs10 with 0, 100, 200, 300, 400 mL per 50 kg seeds were inoculated in each experiment, as well as an additional treatment with commercial product based on B. subtilis. Biomass, nodulation, plant height, internodes, number of pods and number of grains of soybean cultivars were assessed. The inoculation of B. subtilis Bs10 significantly (p < 0.05) improved the agronomic characteristics of both soybean cultivars with an increase in shoot and root biomass, nodulation, phosphorus content, and accumulation of nitrogen in the aerial part. The doses of B. subtilis Bs10 between 200 and 350 mL had significant prominence in increasing the variables studied herein.(AU)

Atualmente, alimentos com baixo teor de agrotóxicos, práticas agrícolas sustentáveis e aumento da produtividade das lavouras têm ganhado destaque no mundo. Inoculantes de rizobactérias são uma alternativa no manejo de grandes lavouras, o que favorece o crescimento sustentável das plantas. Assim, este estudo teve como objetivo avaliar o efeito de doses crescentes do inoculante à base de Rhizobacteria Bacillus subtilis Bs10 em comparação ao produto comercial à base de B. subtilis no desenvolvimento da cultura da soja. Dois experimentos foram conduzidos com as cultivares de soja M8210 IPRO e M8615 IPRO e avaliados nas fases vegetativa e reprodutiva. Doses de B. subtilis Bs10 com 0, 100, 200, 300, 400 mL por 50 kg de sementes foram inoculadas em cada experimento, bem como tratamento adicional com produto comercial à base de B. subtilis. Foram avaliadas biomassa, nodulação, altura de planta, entrenós, número de vagens e número de grãos de cultivares de soja. A inoculação de B. subtilis Bs10 melhorou significativamente (p < 0,05) as características agronômicas de ambas as cultivares de soja com aumento na biomassa da parte aérea e radicular, nodulação, teor de fósforo e acúmulo de nitrogênio na parte aérea. As doses de B. subtilis Bs10 entre 200 e 350 mL destacaram-se significativamente no aumento das variáveis estudadas neste trabalho.(AU)

Production of potato seed tubers under the effect of Trichoderma sp. and rhizobacteria in greenhouse conditions / Produção de tubérculos de batata-semente sob efeito de Trichoderma sp. e rizobactérias em casa de vegetação

Potato (Solanum tuberosum L.) is one of the main crops in the Andean region and due to environmental aspects, the use of biocontrol agents is considered a safe way to produce potato seed tubers.The objective of the study was to evaluate potato inoculation with Trichoderma sp. as a matrix and rhizobacteria Bacillus simplex and Azotobacter sp. on the growth of potato seedlings from in vitro culture, for the production of seed tubers in a greenhouse. The inoculation of microorganisms was carried out in pots, using five potato genotypes for processing. The inoculation treatments were: control, Trichoderma sp., Trichoderma sp. + Azotobacter sp., Trichoderma sp. + Bacillus simplex, Trichoderma sp. + B. simplex + Azotobacter sp. The potato genotypes were cv. Unica (CIP392797.22), cv. Bicentenaria, the advanced clones CIP 396311.1, CIP 399101.1, and the experimental clone UH-09 from the Universidad Nacional José Faustino Sánchez Carrión. A completely randomized design was used, under a factorial arrangement and comparisons between treatments were made at p<0.05. All inoculant treatments exceeded the control in number and weight of tubers per plant as well as in tuber size. Inoculations of Trichoderma sp. alone or with Azotobacter sp. increased plant height, number of leaves per plant and vegetative uniformity; inoculations with the Trichoderma sp. + B. simplex + Azotobacter sp. consortium, improved the dry weight of the foliage, number of shoots per plant and vegetative vigor. There were significant interactions between potato genotypes and inoculant treatments for plant uniformity, vegetative vigor, and the foliage's dry weight. Coinoculation with Trichoderma sp. and some bacterial strains promote the growth of in vitro potato seedlings, increasing the size and weight of the seed tubers and plant biomass, indicating an interrelation between fungi and bacteria that influence the production of potatoes in a greenhouse.(AU)

A batata (Solanum tuberosum L.) é uma das principais culturas da região andina e devido aos aspectos ambientais, o uso de agentes de biocontrole é considerado uma forma segura de produzir tubérculos de batata-semente. O objetivo do trabalho foi avaliar a inoculação de batata com Trichoderma sp. como matriz e rizobactérias Bacillus simplex e Azotobacter sp. sobre o crescimento de mudas de batata provenientes de cultivo in vitro, para produção de tubérculos-semente em casa de vegetação. A inoculação dos microrganismos foi realizada em vasos, utilizando-se cinco genótipos de batata para processamento. Os tratamentos de inoculação foram: testemunha, Trichoderma sp., Trichoderma sp. + Azotobacter sp., Trichoderma sp. + B.simplex, Trichoderma sp. + B.simplex + Azotobacter sp. Os genótipos de batata foram cv. Única (CIP392797.22), cv. Bicentenaria, os clones avançados CIP396311.1, CIP399101.1, e o clone experimental UH-9 da Universidade Nacional José Faustino Sánchez Carrión. O delineamento foi inteiramente casualizado, em esquema fatorial e as comparações entre os tratamentos foram feitas com p<0,05. Todos os tratamentos com inoculantes excederam o controle em número e peso de tubérculos por planta, bem como em tamanho de tubérculo. Inoculações de Trichoderma sp. sozinho ou com Azotobactersp. aumento da altura das plantas, número de folhas por planta e uniformidade vegetativa; inoculações com o Trichoderma sp. + B.simplex + Azotobacter sp. consorcio, melhorou a massa seca da folhagem, número de brotações por planta e vigor vegetativo. Houve interações significativas entre genótipos de batata e tratamentos inoculantes para uniformidade e vigor vegetativo, e para a massa seca da folhagem. Coinoculação com Trichoderma sp. e algumas cepas bacterianas promovem o crescimento de mudas de batata in vitro, aumentando o tamanho e o peso dos tubérculos-semente e da biomassa vegetal, indicando que existe inter-relação entre fungos e bactérias que influenciam a produção de batata em casa de vegetação.(AU)

Refractory feline sporotrichosis: a comparative analysis on the clinical, histopathological, and cytopathological aspects / Esporotricose felina refratária: uma análise comparativa sobre os aspectos clínico, histopatológico e citopatológico

Sporotrichosis is a chronic fungal infection caused by Sporothrix species. The occurrence of cases that are resistant to long-term treatment, especially in the nasal planum of cats, emphasizes the importance of studying its pathogenesis. The purpose of this study was to analyze and compare the inflammatory process of cutaneous lesions of feline refractory sporotrichosis to clinical aspects through cytopathological and histopathological examination. Moreover, the study included 13 cats with cutaneous lesions that had been resistant to itraconazole treatment for more than a year. Cutaneous lesions samples were collected for cytopathological, histopathological, and fungal culture analyses. Tissue fragments were processed and stained with hematoxylin-eosin (HE) and Grocott methenamine silver (GMS). Further, two clinical presentations had the highest occurrence: the localized cutaneous form in animals with good general condition and stable disease (n=9, 69.2%) and the disseminated cutaneous form in cats with poor general condition (n=4, 30.8%). In cats with refractory sporotrichosis, the nasal planum (84.6%) was the most common location of lesions. In the cytopathological study, cats with fewer than two lesions and in good general condition (n=9, 69.2%) showed absence or mild yeast intensity (up to 5 yeasts per field), lower intensity of macrophages and neutrophils, and higher intensity of epithelioid cells, lymphocytes, plasma cells, and eosinophils. On the other hand, (n=4, 30.8%) of the cats with disseminated sporotrichosis and a poor general condition had a marked intensity of yeasts, which were mostly phagocytosed by an increased number of macrophages and neutrophils. Of those animals with good general condition, the majority (n=6, 66.7%) had higher eosinophil intensity. In histopathology, malformed suppurative granuloma was the predominant type (n=9, 69.2%) in feline sporotrichosis lesions, followed by well-formed granulomas (n=4, 30.8%). Malformed granulomas showed mild to moderate fungal intensity (55.6%) in animals with good general condition and localized lesions while marked fungal intensity (44.4%) in cats with the disseminated form of the disease and poor general condition. Well-formed granulomas (n=4, 30.7%) had mild to moderate intensity of fungal load, and 75% of the animals with this type of granuloma had only one lesion and were in good general condition. Long-term itraconazole treatment in these cats with refractory sporotrichosis can keep the infection under control and localized lesions stable; however, fungus reactivation can occur, resulting in an exuberant and inefficient immune response.

A esporotricose é uma infecção fúngica crônica causada por espécies do gênero Sporothrix. A ocorrência de casos refratários ao tratamento de longo prazo, especialmente na região nasal de gatos, alerta para a importância do estudo de sua patogênese. Este estudo teve como objetivo analisar o processo inflamatório das lesões cutâneas de esporotricose felina refratária ao tratamento, por meio de exames citopatológico e histopalógico, e comparar com aspectos clínicos. Treze gatos com lesões cutâneas refratárias ao tratamento com Itraconazol por mais de um ano foram incluídos no estudo. Amostras de lesões cutâneas foram coletadas para análises citopatológicas, histopatológicas e cultura fúngica. Fragmentos de tecidos foram processados e corados pela hematoxilina-eosina (HE) e Prata Metenamina de Grocott (GMS). Duas apresentações clínicas tiveram maior ocorrência: a forma localizada cutânea em animais com bom estado geral e doença estável (n=9, 69,2%); e a forma disseminada cutânea em gatos com estado geral ruim (n=4, 30,8%). A região nasal (84,6%) foi a localização mais frequente das lesões nos gatos com esporotricose refratária ao tratamento. No estudo citopatológico, felinos com menos de duas lesões, e em bom estado geral, (n=9, 69,2%) revelaram ausência ou leve intensidade de leveduras (até 5 leveduras por campo), menor intensidade de macrófagos e neutrófilos, e maior intensidade de células epitelióides, linfócitos, plasmócitos e eosinófilos. Enquanto, (n=4, 30,8%) dos felinos que apresentavam a forma disseminada da esporotricose associada a um estado geral ruim, revelaram acentuada intensidade de leveduras em sua maioria fagocitadas por acentuado número de macrófagos e neutrófilos. Dos animais com bom estado geral, a maioria (n=6, 66,7%) apresentava maior intensidade de eosinófilos. Na histopatologia, o granuloma supurativo mal formado foi o tipo predominante (n=9, 69,2%) nas lesões de esporotricose felina, e (n=4, 30,8%) foram de granulomas bem formados. Granulomas mal formados apresentaram leve a moderada intensidade fúngica (55,6%) nos animais com bom estado geral e lesões localizadas, e acentuada intensidade fúngica (44,4%) nos gatos com a forma disseminada da doença e estado geral ruim. Granulomas bem formados (n=4, 30,7%) apresentaram leve a moderada intensidade de carga fúngica, sendo 75% dos animais com esse tipo de granuloma com apenas uma lesão e bom estadogeral. O tratamento de longo prazo com itraconazol, nesses gatos com esporotricose refratária, pode manter a infecção controlada e lesões localizadas estáveis de forma temporária, contudo a reativação do fungo pode ocorrer levando a uma resposta imunológica exuberante e ineficiente.