Activity of biogenic silver nanoparticles in planktonic and biofilm-associated Corynebacterium pseudotuberculosis.

Corynebacterium pseudotuberculosis is a gram-positive bacterium and is the etiologic agent of caseous lymphadenitis (CL) in small ruminants. This disease is characterized by the development of encapsulated granulomas in visceral and superficial lymph nodes, and its clinical treatment is refractory to antibiotic therapy. An important virulence factor of the Corynebacterium genus is the ability to produce biofilm; however, little is known about the characteristics of the biofilm produced by C. pseudotuberculosis and its resistance to antimicrobials. Silver nanoparticles (AgNPs) are considered as promising antimicrobial agents, and are known to have several advantages, such as a broad-spectrum activity, low resistance induction potential, and antibiofilm activity. Therefore, we evaluate herein the activity of AgNPs in C. pseudotuberculosis, through the determination of minimum inhibitory concentration (MIC), minimum bactericidal concentration (MBC), antibiofilm activity, and visualization of AgNP-treated and AgNP-untreated biofilm through scanning electron microscopy. The AgNPs were able to completely inhibit bacterial growth and inactivate C. pseudotuberculosis at concentrations ranging from 0.08 to 0.312 mg/mL. The AgNPs reduced the formation of biofilm in reference strains and clinical isolates of C. pseudotuberculosis, with interference values greater than 80% at a concentration of 4 mg/mL, controlling the change between the planktonic and biofilm-associated forms, and preventing fixation and colonization. Scanning electron microscopy images showed a significant disruptive activity of AgNP on the consolidated biofilms. The results of this study demonstrate the potential of AgNPs as an effective therapeutic agent against CL.

[Síndrome de fiebre periódica, adenitis, faringitis y aftosis en un preescolar con síndrome de Asperger: reporte de caso].

Background: Autoinflammatory diseases are a heterogeneous group of pathologies whose prevalence is undefined to date. Within these, the Periodic Fever Syndrome, Adenitis, Pharyngitis and Aphthosis (PFAPA) is determined as the most common and the most studied. Neurodevelopmental and behavioral disor- ders, including those of the Autism Spectrum, are especially prevalent in patients with allergic and autoimmune diseases, however, there is little documented regarding their relationship with autoinflammation. Case report: The case of a patient with Asperger Syndrome with subsequent onset of this autoinflammatory disease is shown. Two weekly blood counts and complete serum immunoglobulins are obtained, which do not show a decrease in neutrophil counts or sublevels or supralevels in immunoglobulins. Conclusion: When he returned to his follow-up appointment, he reported a feverish peak 2 days before, which ceased with the administration of the steroid, and if there was presence of oral ulcers on this occasion, fulfilling the modified Marshall criteria. Likewise, a great change is noted in their development and cooper- ation in exploration. The duality of this autoinflammatory syndrome in a patient with Asperger Syndrome is determined.

Antecedentes: Las enfermedades autoinflamatorias son un grupo heterogéneo de patologías cuya prevalencia es indefinida hasta nuestra actualidad. Dentro de estas, el Síndrome de Fiebre Periódica, Adenitis, Faringitis y Aftosis (PFAPA, por sus siglas en inglés) se determina como la más común y de las mayormente es- tudiadas. Las alteraciones del neurodesarrollo y la conducta, entre estas las del Espectro Autista, son especialmente prevalentes en pacientes con enfermedades alérgicas y autoinmunes, sin embargo, hay poco documentado en cuanto a su relación con la autoinflamación. Reporte de caso: Se muestra el caso de un paciente con Síndrome de Asperger con debut posterior de esta enfermedad autoinflamatoria. Se obtienen 2 biometrías hemáticas semanales e inmunoglobulinas séricas completas, las cuales no muestran descenso en conteos de neutrófilos ni infraniveles ni supraniveles en las inmunoglobulinas. Conclusión: Al reacudir a su cita de control refiere pico febril 2 días previos, el cual cesó con la administración del esteroide, ya si presencia de úlceras orales en esta ocasión, cumpliendo los criterios modificados de Marshall. Por igual, se nota un gran cambio en su desenvolvimiento y cooperación a la exploración. Se determina la dualidad de este síndrome autoinflamatorio en un paciente con Síndrome de Asperger.

A serum NMR metabolomic analysis of the Corynebacterium pseudotuberculosis infection in goats.

Caseous lymphadenitis (CLA), an infectious disease caused by Corynebacterium pseudotuberculosis in small ruminants, is highly prevalent worldwide. Economic losses have already been associated with the disease, and little is known about the host-pathogen relationship associated with the disease. The present study aimed to perform a metabolomic study of the C. pseudotuberculosis infection in goats. Serum samples were collected from a herd of 173 goats. The animals were classified as controls (not infected), asymptomatic (seropositives but without detectable CLA clinical signs), and symptomatic (seropositive animals presenting CLA lesions), according to microbiological isolation and immunodiagnosis. The serum samples were analyzed using nuclear magnetic resonance (1H-NMR), nuclear Overhauser effect spectroscopy (NOESY), and Carr-Purcell-Meiboom-Gill (CPMG) sequences. The NMR data were analyzed using chemometrics, and principal component analysis (PCA) and partial least square discriminant analysis (PLS-DA) were performed to discover specific biomarkers responsible for discrimination between the groups. A high dissemination of the infection by C. pseudotuberculosis was observed, being 74.57% asymptomatic and 11.56% symptomatic. In the evaluation of 62 serum samples by NMR, the techniques were satisfactory in the discrimination of the groups, being also complementary and mutually confirming, demonstrating possible biomarkers for the infection by the bacterium. Twenty metabolites of interest were identified by NOESY and 29 by CPMG, such as tryptophan, polyunsaturated fatty acids, formic acid, NAD+, and 3-hydroxybutyrate, opening promising possibilities for the use of these results in new therapeutic, immunodiagnosis, and immunoprophylactic tools, as well as for studies of the immune response against C. pseudotuberculosis. KEY POINTS: • Sixty-two samples from healthy, CLA asymptomatic, and symptomatic goats were screened • Twenty metabolites of interest were identified by NOESY and 29 by CPMG • 1H-NMR NOESY and CPMG were complementary and mutually confirming.

Potential in vitro action of an adenosine analog and synergism with penicillin against Corynebacterium pseudotuberculosis.

Caseous lymphadenitis is a well-known disease caused by Corynebacterium pseudotuberculosis affecting small ruminants with small significance to human health because of its minor zoonotic potential. In both cases, few treatment options are available and conventional antimicrobial therapy is commonly refractory due to development of pyogranulomatous reactions, bringing great interest in discovering novel therapeutics for more suitable approaches. Dideoxynucleotides presented antibacterial action against various bacteria but were never described for C. pseudotuberculosis. Hypothesizing the antimicrobial action of 2',3'-dideoxiadenosine (ddATP) against C. pseudotuberculosis, we performed for the first time an investigation of its minimum inhibitory concentration (MIC) and minimum bactericidal concentration (MBC) in the ATCC® 19,410 strain and a well-characterized clinical isolate of C. pseudotuberculosis. We also assessed potential synergism with penicillin. ddATP showed a growth delay effect for C. pseudotuberculosis at 2 µmol/mL and a MIC and MBC of 4 µmol/mL against the ATCC® 19,410 strain, but not for the clinical strain. An antimicrobial effect was observed when using concentrations lower than the MIC of ddATP associated with penicillin for both strains tested. Our data suggest the potential of nucleotide analogs, especially adenosine, and its combination with penicillin, as a possible novel treatment for C. pseudotuberculosis-induced infections, and contributes with knowledge regarding alternative drugs to treat C. pseudotuberculosis infections.

Bioinformatic Approach of B and T Cell Epitopes of PLD and CP40 Proteins of Corynebacterium pseudotuberculosis ovis Mexican Isolate 2J-L towards a Peptide-Based Vaccine.

Mapping B and T cell epitopes constitutes an important action for peptide vaccine design. PLD and CP40 virulence factors of Corynebacterium pseudotuberculosis biovar ovis, a causal agent of Caseous Lymphadenitis, have been evaluated in a murine model as good candidates for vaccine development. Therefore, the goal of this work was to in silico analyze B and T cell epitopes of the PLD and CP40 proteins of a Mexican isolate of Corynebacterium pseudotuberculosis ovis. The Immune Epitope Data Base and Resource website was employed to predict the linear and conformational B-cell, T CD4+, and T CD8+ epitopes of PLD and CP40 proteins of Corynebacterium pseudotuberculosis ovis Mexican strain 2J-L. Fifty B cell epitopes for PLD 2J-L and forty-seven for CP40 2J-L were estimated. In addition, T CD4+ and CD8+ cell epitopes were predicted for PLD 2J-L (MHC I:16 epitopes, MHC II:10 epitopes) and CP40 2J-L (MHC I: 15 epitopes, MHC II: 13 epitopes). This study provides epitopes, paying particular attention to sequences selected by different predictor programs and overlap sequences as B and T cell epitopes. PLD 2J-L and CP40 2J-L protein epitopes may aid in the design of a promising peptide-based vaccine against Caseous Lymphadenitis in Mexico.

Morganella sp. infection in scent gland of corn snake (Pantherophis guttatus) - case report / Infecção por Morganella sp. em glândula de cheiro de corn snake (Pantherophis guttatus) - relato de caso

The corn snake (Pantherophis guttatus) is a non-venomous snake from the Colubridae family. The pair of scent glands is indispensable in the physiology of reproduction and defense of the species. This structure is located caudal to the cloaca. It is responsible for releasing a thick material based on lipids containing pheromones that perform functions associated with animal behavior and survival. This work aims to report infection by Morganella sp. in a scent gland in a female corn snake, evidencing the diagnosis and treatment. An increase in volume was seen in the final third, proximal to the cloaca, firm consistency, immobile, and approximately 4 cm long and painful to touch. Samples of the material were collected for bacterial culture and antibiogram examination, being positive for bacteria of the genus Morganella. Based on the result of the antibiogram, it was possible to determine the most appropriate therapeutic protocol, with the referral to perform the surgical procedure to remove the tissue compromised by the infection, remaining stable until the removal of the stitches 40 days after the procedure, with total surgical wound healing.(AU)

A corn snake (Pantherophis guttatus) é uma serpente não peçonhenta, da família Colubridae. Na fisiologia da reprodução e defesa da espécie, o par de glândulas de cheiro é indispensável. Essa estrutura está localizada caudal à cloaca e é responsável pela liberação de um material espesso à base de lipídeos, contendo feromônios que exercem funções associadas ao comportamento animal e à sua sobrevivência. O objetivo desse trabalho é relatar uma infecção por Morganella sp. em glândula de cheiro em uma fêmea de corn snake, evidenciando o diagnóstico e tratamento. Observou-se um aumento de volume no terço final, proximal a cloaca, de consistência firme, imóvel e com aproximadamente 4 cm de comprimento e sensibilidade dolorosa ao toque. Foram coletadas amostras do material para cultura bacteriana e exame de antibiograma, sendo positivo para bactérias do gênero Morganella. Com base no resultado do antibiograma, foi possível determinar o protocolo terapêutico mais adequado, como o encaminhamento para realização do procedimento cirúrgico para remoção do tecido comprometido pela infecção, se mantendo estável até a remoção dos pontos, 40 dias após o procedimento, com total cicatrização da ferida cirúrgica.(AU)

General aspects of the production and management systems and their relationship with Corynebacterium pseudotuberculosis infection and occurrence of mastitis of white Morada Nova sheep herds in semi-arid region / Aspectos gerais do sistema de produção e manejo e sua relação com a infecção por Corynebacterium pseudotuberculosis e ocorrência de mastite de rebanhos da raça Morada Nova branca em região semiárida

This study aimed to characterize the production and management systems of white Morada Novasheep herds and to present data on the occurrence of caseous lymphadenitis (CLA) and clinical mastitis (CM) in a population maintained in a semi-arid region. Blood samples were collected from adult females from herds of the Brazilian northeastern region. The first venous blood sample was used for a complete blood examination. The second blood sample was used to detect the presence of anti-C. pseudotuberculosis specific antibodies. The production and management systems of the herds were characterized using questionnaires, divided into three blocks of indicators: Facilities; Characterization of the production system; Health management. Body weight and hematological results were submitted to an analysis of variance, on which the fixed effect was clinical diagnosis, and the principal component analysis was performed to determine the main variables. For CLA, three herds stood out as having the highest rates of seropositivity (85.71, 79.41 and 60.45%). The same herds also had the highest occurrences of CM (9.30, 8.82 and 5.71%). The main variables that showed differences according the diagnosis of both diseases were blood granulocytes and lymphocytes. The study made it possible to observe the relationship between the best sanitary practices and the lowest occurrence rates of both CLA and CM. The diseases present considerable occurrence in sheep populations

Este estudo teve como objetivo caracterizar os sistemas de produção e manejo de rebanhos de ovelhas Morada Nova da variedade branca e apresentar dados sobre a ocorrência de linfadenite caseosa (CLA) e mastite clínica (MC) em uma população mantida em região semiárida. Amostras de sangue foram coletadas de fêmeas adultas de rebanhos da região nordeste do Brasil. A primeira amostra de sangue venoso foi utilizada para um exame de sangue completo. A segunda amostra de sangue foi utilizada para detectar a presença de anticorpos anti-C. anticorpos específicos de pseudotuberculosis. Os sistemas de produção e manejo dos rebanhos foram caracterizados por meio de questionários, divididos em três blocos de indicadores: Instalações; Caracterização do sistema de produção; Gestão de saúde do rebanho. O peso corporal e os resultados hematológicos foram submetidos a uma análise de variância, na qual o efeito fixo foi o diagnóstico clínico, e a análise de componentes principais foi realizada para determinar as variáveis principais. Para o CLA, três rebanhos se destacaram com os maiores índices de soropositividade (85,71, 79,41 e 60,45%). Os mesmos rebanhos também apresentaram as maiores ocorrências de MC (9,30, 8,82 e 5,71%). As principais variáveis que apresentaram diferenças de acordo com o diagnóstico de ambas as doenças foram granulócitos e linfócitos. O estudo possibilitou observar a relação entre as melhores práticas sanitárias e as menores taxas de ocorrência tanto de CLA quanto de MC. As doenças apresentam ocorrência considerável nas populações de ovinos

Hematological and clinical biochemistry profiles in Canindé goats infected by Corynebacterium pseudotuberculosis and bred in a tropical semi-arid region.

Caseous lymphadenitis (CLA), an infectious disease caused by Corynebacterium pseudotuberculosis in goats and sheep, is highly prevalent worldwide and is characterized by economic losses in small ruminant production. Currently available techniques for clinical and laboratory diagnosis of the disease lack market availability and/or sensitivity, and therefore, infected animals can remain in the herd, serving as a source of infection for other animals. The present study aimed to verify hematological and clinical biochemistry changes in goats naturally infected by C. pseudotuberculosis. One hundred seventy-three Canindé goats were included in this study, from which blood samples and caseous lesions were collected. The animals were classified as uninfected, asymptomatic, and symptomatic according to microbiological isolation and serological assays. A high dissemination of the infection was observed in the herd, with 86.13% of positive animals, being 74.57% asymptomatic and 11.56% symptomatic. In the hemogram and clinical biochemistry analyses, the only statistical difference found was a higher level of serum urea in asymptomatic individuals than in non-infected animals. In addition, this study points to the possibility of chronic CLA being potentially reflected in hepatic and renal biochemical markers.

Assessment of rSodC, rPknG, rNanH, and rSpaC as Antigens for Diagnostic Tools Against Caseous Lymphadenitis.

Corynebacterium pseudotuberculosis is a bacillus that causes caseous lymphadenitis in small ruminants, leading to great losses to rural producers; thus, an efficient diagnosis is necessary for using disease control measures. This study aimed to evaluate the antigenic potential of four C. pseudotuberculosis recombinant proteins (rSodC, rPknG, rNanH, and rSpaC) against sera of goat and sheep experimentally infected with one of three different C. pseudotuberculosis strains. Goats were infected with CAP76 or CAP21 strain (n = 10), sheep with VD57 strain (n = 6), and a group of not-infected animals (goats and sheep) were kept as a healthy control (healthy n = 12). Sera were collected at 0, 14, 60, 90, 180, or 190 days after inoculation for antigenicity testing using Western blotting and enzyme-linked immunosorbent assay (ELISA) techniques. Cross-reactivity tests with recombinant proteins were performed in goat serum experimentally vaccinated with Nocardia sp. or Rhodococcus equi bacterin. The rSodC protein showed discriminatory antigenic reactivity with a statistically significant difference against three different C. pseudotuberculosis strains evaluated in goats and sheep samples, while rPknG showed statistical significance only against two C. pseudotuberculosis strains evaluated in goats. rSodC was proved to be a strong candidate as a tool for diagnosis of C. pseudotuberculosis infection, once it was able to recognize antibodies against all strains evaluated in goats and sheep.

Linfadenitis por Mycobacterium en pacientes inmunocompetentes y viviendo con VIH/sida / Lymphodenitis by Mycobacterium in immunocompetent and living with HIV/AIDS patients

Introducción: La coinfección del virus de inmunodeficiencia humana (VIH) y la tuberculosis ha alterado su presentación histológica, esto es particularmente frecuente en las linfadenitis. Objetivos: Realizar la caracterización etiológica de linfadenopatías producidas por el género Mycobacterium, destacar la importancia del diagnóstico precoz de esta enfermedad para evitar diseminación de la infección, tanto en pacientes inmunocompetentes como inmunodeficientes, específicamente con VIH/sida. Método: Se realizó estudio descriptivo-prospectivo entre enero de 2017 y enero de 2019. Durante este período se recibieron 5640 muestras, de estas 81 obtenidas a partir de tejido ganglionar; la toma de muestra mayoritariamente fue quirúrgica 74 (91,35 por ciento) y 7 (8,64 por ciento) por biopsia aspirativa (BAAF). Del total de muestras, 60 (74,07 por ciento) procedían de pacientes con VIH/sida, las muestras se descontaminaron por el método de ácido sulfúrico al 4 por ciento, se cultivaron en medio sólido Löwenstein-Jensen e incubaron a 37°C. Se realizaron lecturas semanalmente. Para identificar Mycobacterium tuberculosis se realizó la prueba rápida comercial inmunocromatográfica SD TB AgMPT64. Resultados: De 81 muestras analizadas se obtuvieron 22 (27,16 por ciento) aislamientos, 16 (72,72 por ciento) de Mycobacterium tuberculosis, y 6 (27,27 por ciento) de especies no tuberculosas. De estas, 18 (81,81 por ciento) procedían de pacientes con VIH/sida. Conclusión: Por todo lo antes expuesto es importante la vigilancia diagnóstica en este tipo de infección extrapulmonar, tanto para M. tuberculosis como para otras especies no tuberculosas y poder comenzar tempranamente el tratamiento específico evitando la diseminación de la infección, pues esta puede tener consecuencias fatales, sobre todo en pacientes con algún tipo de inmunosupresión, como aquellos con VHI/sida. Si un paciente mantiene fiebre prolongada, con linfadenopatías, sin síntomas respiratorios y no responde a los tratamientos con antibióticos, es necesario pensar en este tipo de infección(AU)

Introduction: The coinfection of human immunodeficiency virus (HIV) and tuberculosis has altered its histological presentation; this is particularly frequent in lymphadenitis. Objective: To carry out the etiological characterization of lymphadenopathies produced by the genus Mycobacterium, highlighting the importance of early diagnosis of this disease to avoid dissemination of the infection, both in immunocompetent and immunodeficient patients, specifically HIV / AIDS. Methods: A descriptive-prospective study was carried out between January 2017 - January 2019. During this period, 5640 samples were received, of these 81 obtained from lymph node tissue, the sample collection was mostly surgical 74 (91.35 percent) and 7 (8.64 percent) by aspiration biopsy (BAAF). Of the total samples, 60 (74.07 percent) were from HIV / AIDS patients, the samples were decontaminated by the 4 percent sulfuric acid method and cultured in solid Löwenstein-Jensen medium and incubated at 370C, the readings were made weekly. For the identification of Mycobacterium tuberculosis, the commercial SD TB AgMPT64 immunochromatographic rapid test was performed. Results: Of 81 samples analyzed, 22 (27.16 percent) isolates were obtained, 16 (72.72 percent) of Mycobacterium tuberculosis (MTB), and 6 (27.27 percent) of non-tuberculous species, of these 18 (81.81%) were from HIV / AIDS patients. Conclusion: For all the above, diagnostic surveillance is important in this type of extrapulmonary infection, both for M tuberculosis and for other non-tuberculous species and to be able to start specific treatment early, avoiding the spread of the infection, since it can have fatal consequences on all in patients with some type of immunosuppression, such as HIV/AIDS. If a patient maintains a prolonged fever, with lymphadenopathy, without respiratory symptoms and does not respond to antibiotic treatment, it is necessary to consider this type of infection(AU)

Periodic fever, aphthous stomatitis, pharyngitis and cervical adenitis syndrome in children: a brief literature review.

OBJECTIVE: To describe clinical, diagnostic and therapeutic characteristics of the periodic fever, aphthous stomatitis, pharyngitis and cervical adenitis (PFAPA) syndrome. DATA SOURCE: Literature review in the PubMed database by using specific descriptors to identify all articles published in the English language in the last three years; 38 articles were found. After performing selection of titles and abstract analysis, 13 out of the 38 articles were fully read. Relevant studies found in the references of the reviewed articles were also included. DATA SYNTHESIS: The PFAPA syndrome (Periodic Fever, Aphthous Stomatitis, Pharyngitis and cervical Adenitis) is a medical condition grouped among the periodic fever syndromes. The etiology is uncertain, but possibly multifactorial, and its symptoms are accompanied by recurrent febrile episodes although weight and height development are preserved. It is a self-limiting disease of benign course with remission of two to three years without significant interference in the patient's overall development. Treatment consists of three pillars: interruption of febrile episodes, increase in the interval between episodes, and remission. CONCLUSIONS: Despite several attempts to establish more sensitive and specific criteria, the diagnosis of PFAPA syndrome is still clinical and reached by exclusion, based on the modified Marshall's criteria. The most common pharmacological options for treatment include prednisolone and betamethasone; colchicine may be used as prophylaxis, and surgical treatment with tonsillectomy can be considered in selected cases.

Chemokine production induced by Corynebacterium pseudotuberculosis in a murine model.

Corynebacterium pseudotuberculosis is the etiological agent of caseous lymphadenitis. The main clinical sign of this disease is the development of granulomas, especially in small ruminants; however, the pathways that are involved in the formation and maintenance of these granulomas are unknown. Cytokines and chemokines are responsible for the migration of immune cells to specific sites and tissues; therefore, it is possible that chemokines participate in abscess formation. This study aimed to evaluate the induction of chemokine production by two C. pseudotuberculosis strains in a murine model. A highly pathogenic (VD57) and an attenuated (T1) strain of C. pseudotuberculosis, as well as somatic and secreted antigens derived from these strains, was used to stimulate murine splenocytes. Then, the concentrations of the chemokines CCL-2, CCL-3, CCL-4, and CCL-5 and the cytokines IL-1 and TNF were measured in the culture supernatants. The VD57 strain had a higher ability to stimulate the production of chemokines when compared to T1 strain, especially in the early stages of stimulation, which can have an impact on granuloma formation. The T1 lysate antigen was able to stimulate most of the chemokines studied herein when compared to the other antigenic fractions of both strains. These results indicate that C. pseudotuberculosis is a chemokine production inducer, and the bacterial strains differ in their induction pattern, a situation that can be related to the specific behavior of each strain.

Granulomatous lymphadenitis in a dog caused by Mycobacterium intracellulare

Background: Mycobacteriosis is caused by bacteria belonging to the genus Mycobacterium, with considerable zoonoticpotential and risk to public health. Infection in dogs is rare and is usually associated with immunosuppression, resultingfrom eating meat or contact with contaminated soil or fomites. Dogs are also known as potential sources for the spread ofatypical tuberculosis in humans and other animals. This paper aims to describe the clinical, cytological, histopathological,and molecular findings of a male canine seen at University Veterinary Hospital of Cuiabá, Mato Grosso, with generalizedlymphadenomegaly associated Mycobacterium intracellulare infection.Case: A 2-year-old male Lhasa Apso dog was referred to the University Veterinary Hospital in Cuiabá city, located in theMidwest region of Brazil. The patient had a history of intermittent claudication of the left pelvic limb for approximately6 months and lymphadenomegaly with progression for approximately 2 months. The dog had wheezing and generalizedlymphadenopathy (submandibular, axillary, and popliteal lymph nodes); cryptorchidism was also observed. A completeblood count revealed nonspecific results, and in the serum biochemical profile, the values of urea, creatinine, albumin, andalanine aminotransferase were within the reference range. No changes were observed on the radiography of the femurotibiopatellar joints. Considering the generalised lymphadenopathy, fine needle aspiration cytology and histopathologicalexamination through biopsy of the lymph nodes was performed. On the cytology and histopathology, numerous negativeimages of moderately refringent bacillary structures distending the cytoplasm from the macrophages was found. Thesamples were also subjected to special Ziehl-Neelsen staining, which confirmed an accentuated and diffuse granulomatouslymphadenitis associated with alcohol...(AU)

Granulomatous lymphadenitis in a dog caused by Mycobacterium intracellulare

Background: Mycobacteriosis is caused by bacteria belonging to the genus Mycobacterium, with considerable zoonoticpotential and risk to public health. Infection in dogs is rare and is usually associated with immunosuppression, resultingfrom eating meat or contact with contaminated soil or fomites. Dogs are also known as potential sources for the spread ofatypical tuberculosis in humans and other animals. This paper aims to describe the clinical, cytological, histopathological,and molecular findings of a male canine seen at University Veterinary Hospital of Cuiabá, Mato Grosso, with generalizedlymphadenomegaly associated Mycobacterium intracellulare infection.Case: A 2-year-old male Lhasa Apso dog was referred to the University Veterinary Hospital in Cuiabá city, located in theMidwest region of Brazil. The patient had a history of intermittent claudication of the left pelvic limb for approximately6 months and lymphadenomegaly with progression for approximately 2 months. The dog had wheezing and generalizedlymphadenopathy (submandibular, axillary, and popliteal lymph nodes); cryptorchidism was also observed. A completeblood count revealed nonspecific results, and in the serum biochemical profile, the values of urea, creatinine, albumin, andalanine aminotransferase were within the reference range. No changes were observed on the radiography of the femurotibiopatellar joints. Considering the generalised lymphadenopathy, fine needle aspiration cytology and histopathologicalexamination through biopsy of the lymph nodes was performed. On the cytology and histopathology, numerous negativeimages of moderately refringent bacillary structures distending the cytoplasm from the macrophages was found. Thesamples were also subjected to special Ziehl-Neelsen staining, which confirmed an accentuated and diffuse granulomatouslymphadenitis associated with alcohol...

Molecular characterization of Corynebacterium pseudotuberculosis, C. silvaticum, and C. auriscanis by ERIC-PCR / Caracterização molecular de Corynebacterium pseudotuberculosis, C. silvaticum e C. auriscanis pelo ERIC-PCR

The aims of the present study were (i) to genotype Corynebacterium pseudotuberculosis, C. silvaticum, and C. auriscanis strains using enterobacterial repetitive intergenic consensus (ERIC-PCR), and (ii) to analyze the epidemiological relationships among isolates according to biovar (Equi and Ovis), species, host, and geographical origin of the C. pseudotuberculosis strains. Sixty-eight C. pseudotuberculosis, nine C. silvaticum, and one C. auriscanis, C. pseudotuberculosis ATCC® 19410™ strain and the attenuated C. pseudotuberculosis 1002 vaccinal strain were fingerprinted by ERIC 1+2-PCR. Field strains were isolated from various hosts (cattle, buffaloes, sheep, goats, horses, dogs, and pigs) in six countries (Mexico, Portugal, Brazil, Equatorial Guinea, Egypt, and Israel). High genetic diversity was found among the studied Corynebacterium spp. isolates, clustering in 24 genotypes with a Hunter & Gaston diversity index (HGDI) of 0.937. The minimal spanning tree of Corynebacterium spp. revealed three clonal complexes, each associated with one bacterial species. Twenty-two genotypes were observed among C. pseudotuberculosis isolates, with an HGDI of 0.934. Three major clonal complexes were formed at the minimal spanning tree, grouped around the geographic origin of C. pseudotuberculosis isolates. These results reinforce the high typeability, epidemiological concordance, and discriminatory power of ERIC-PCR as a consistent genotyping method for C. pseudotuberculosis, which could be useful as an epidemiological tool to control caseous lymphadenitis. Moreover, our results also indicate the potential of ERIC 1+2-PCR for the genotyping of other species of Corynebacterium other than C. pseudotuberculosis.

Os objetivos do presente estudo foram (i) genotipar amostras de Corynebacterium pseudotuberculosis, C. silvaticum e C. auriscanis usando Enterobacterial Repetitive Intergenic Consensus (ERIC-PCR), bem como (ii) analisar as relações epidemiológicas entre os isolados de acordo com biovar (Equi e Ovis), espécie, hospedeiro e origem geográfica das amostras de C. pseudotuberculosis. Sessenta e oito isolados de C. pseudotuberculosis, nove C. silvaticum, um C. auriscanis, C. pseudotuberculosis ATCC® 19410 ™ e a amostra vacinal atenuada C. pseudotuberculosis 1002 foram tipificadas por ERIC 1 + 2-PCR. As amostras de campo foram isoladas de diferentes hospedeiros (bovinos, búfalos, ovinos, caprinos, equinos, cães e suínos) em seis países (México, Portugal, Brasil, Guiné Equatorial, Egito e Israel). Uma alta diversidade genética foi observada entre os isolados de Corynebacterium spp., agrupados em vinte e quatro genótipos com um índice de diversidade Hunter & Gaston (HGDI) de 0,937. A análise da minimal spanning tree (MST) de Corynebacterium spp. revelou três complexos clonais, cada um associado a uma espécie bacteriana. Vinte e dois genótipos foram observados entre isolados de C. pseudotuberculosis, com um HGDI de 0,934. Na análise da MST, três grandes complexos clonais foram formados, agrupando-se em torno da origem geográfica dos isolados de C. pseudotuberculosis. Esses resultados reforçam a alta tipabilidade, concordância epidemiológica e poder discriminatório do ERIC-PCR como método consistente de genotipagem para C. pseudotuberculosis, podendo ser útil como ferramenta epidemiológica no controle da linfadenite caseosa. Além disso, os resultados também indicam o grande potencial de ERIC 1 + 2-PCR para genotipagem de espécies do gênero Corynebacterium além de C. pseudotuberculosis.

Peptide vaccines designed with the aid of immunoinformatic against Caseous Lymphadenitis promotes humoral and cellular response induction in mice.

Caseous Lymphadenitis (CLA) is a chronic disease that affects also small ruminants. CLA is caused by Corynebacterium pseudotuberculosis and is responsible for high economic losses due to the formation of superficial and visceral granulomas, the latter is considered as asymptomatic CLA causing high levels of dissemination. Several vaccination strategies, in which the use of synthetic peptides stands out. Thus, this work aimed to evaluate the protective potential of peptide vaccines designed to determine the immunodominant epitopes of CP40 against CLA in mice. The animals were divided into eight groups separated in controls (G1-PBS, G2-Saponin and G9-rCP40) and experimental (G3-pep1, G4- pep2, G5-pep3, G6-pep4, G7-pep5 and G8-pep6), these were vaccinated on days 0 and 15 by a subcutaneous route. 60 days after the first immunization, all animals were challenged with C. pseudotuberculosis. On days 0, 15, 60, and 120 after the first immunization, blood samples were taken to measure immunoglobulins. On the same day of the challenge, the splenocytes were isolated and assayed for the production of IL-2, IL-4, IL-6, IFN-γ, TNF-α, IL-17, and IL-10. After vaccinations, the animals were challenged and all of them were affected by the disease which led to their death. The G6 and G8 groups provided 10% protection and the G7 provided 20%. The G3 and G4 groups provided 30% and 40% protection respectively. The peptides showed the production of Total IgG antibodies and cytokines (IL-2, IL-4, IL-6, IFN-γ, and TNF-α), indicating a possible activation of the Th1 type response. However, groups G3, G5, G6, and G8 showed production of IL-17. None of the study groups showed IL-10 production. The immunogenicity of the peptides was not enough to protect these animals and it is believed that the use of adjuvants based on PAMPs may improve the immune response offered by these peptides.

In silico analyses and design of a chimeric protein containing epitopes of SpaC, PknG, NanH, and SodC proteins for the control of caseous lymphadenitis.

Caseous lymphadenitis (CLA) is an infectious disease that affects goats and sheep causing drastic impacts on milk and meat production and is caused by Corynebacterium pseudotuberculosis. The disease can be prevented through vaccination but currently, vaccines demonstrate limited efficacy consequently leading to a need for the development of new ones. Here, we described the in silico development of a new chimeric protein constructed with epitopes identified from the sequences of the genes nanH, pknG, spaC, and sodC, previously described as potential vaccinal targets against C. pseudotuberculosis. The chimera was expressed, purified, and its immunogenicity was evaluated using sera of immunized mice. Results indicate the chimeric protein was able to stimulate antibody production. Additionally, analysis using serum from naturally infected goats showed that the protein is recognized by sera from these animals, indicating the possibility for using this chimera in new diagnostic methods. KEY POINTS: • The chimera was expressed with 52 kDa and a yield of 7 mg/L after purification. • The chimera was recognized by the sera of animals immunized with this formulation. • Chimera reacted with the serum of goats naturally infected with C. pseudotuberculosis.