Chronic Diarrhea Due to Intestinal Spirochaetosis in an HIV-Positive Patient: Case Report and Literature Review

Intestinal spirochaetosis refers to the colonization of the colorectal epithelium by anaerobic spirochetes of the genus Brachyspira. It is more frequently identified in developing countries or when sanitation conditions are suboptimal. It can affect anyone, although it is more common in gay men, particularly those infected with the human immunodeficiency virus (HIV). Its most common symptom is chronic watery diarrhea. A colonoscopy may be expected. We present the clinical case of a young male patient with chronic diarrhea of several years of evolution resolved by specific treatment. In addition, a review of the literature on this disease is provided.

La espiroquetosis intestinal se define como la colonización del epitelio colorrectal por espiroquetas anaerobias del género Brachyspira. Se identifica con mayor frecuencia en países en vías de desarrollo o cuando las condiciones de salubridad son subóptimas. Puede afectar a cualquier persona, aunque es más común en hombres homosexuales particularmente infectados por virus de la inmunodeficiencia humana (VIH). Su síntoma más frecuente es la diarrea crónica acuosa. La colonoscopia puede ser normal. Se presenta el caso clínico de un paciente masculino joven con diarrea crónica de varios años de evolución que se resuelve al brindar tratamiento específico. Además, se realiza, una revisión de la literatura sobre esta enfermedad.

Diagnosis of Brachyspira pilosicoli, Brachyspira hyodysenteriae and Brachyspira intermedia in hens and laying hens in the western region of Paraná through bacterial isolation and identification in qPCR / Diagnóstico de Brachyspira pilosicoli, Brachyspira hyodysenteriae e Brachyspira intermedia em aves de postura e matrizes de corte na região oeste do Paraná através do isolamento bacteriano e identificação na qPCR

Bacteria of the genus Brachyspira can cause enteric diseases in poultry causing a decrease in productivity. The occurrence of this disease in chickens has already been verified in countries such as Australia, Italy, and the United States, but in Brazil, until now, epidemiological studies about Brachyspira sp. frequency were only carried out on pig farms. The objective of this study was to evaluate the presence of bacteria of the genus Brachyspira sp. through isolation and confirmation of the species Brachyspira pilosicoli, Brachyspira hyodysenteriae and Brachyspira intermedia using the qPCR technique. Samples from 110 hens aged from 35 to 82 weeks were collected, 40 were from commercial egg farms and 70 were from laying hens matrices. For the first evaluation, bacterial isolation was performed from the feces. Positive samples were submitted to qPCR to identify the three species proposed. Cecum fragments of the birds were collected and fixed in formaldehyde for histological evaluation and counting of goblet cells. Of the 110 samples, 48 characteristic isolates of Brachyspira (43.6%) were obtained and of these in qPCR 13 identified as B. hyodysenteriae (11.8%) and 5 all from the same farm as Brachyspira intermedia (4.5%), 2 samples were positive for both agents (1.8%) and 28 were not characterized by qPCR (25.5%). None histopathological lesions were observed in the chicken cecum and no significant statistical difference was noticed in the count of goblet cells of the positive hens. It can be evidenced by the occurrence of Brachyspira sp. in laying farms and hens in Brazil, with special relevance to Brachyspira intermedia that can be potentially pathogenic for these animals.(AU)

Bactérias do gênero Brachyspira podem ocasionar enfermidades entéricas em aves acarretando a queda de produtividade. A ocorrência desta enfermidade em galinhas já foi verificada em países como a Austrália, Itália e Estados Unidos, porém no Brasil, até o momento, trabalhos epidemiológicos sobre a frequencia de Brachyspira sp. só foram realizados em granjas de suínos. O objetivo deste estudo foi avaliar a presença de bactérias do gênero Brachyspira sp. através do isolamento e confirmação das espécies Brachyspira pilosicoli, Brachyspira hyodysenteriae e Brachyspira intermedia utilizando a técnica de qPCR. Foram coletadas amostras de 110 aves com idade entre 35 e 82 semanas, sendo 40 de granjas de postura comercial e 70 de granjas de matrizes de corte. Para avaliação primeiramente procedeu-se o isolamento bacteriano a partir das fezes. As amostras positivas foram submetidas a qPCR para identificação das três espécies propostas. Fragmentos de ceco das aves foram coletados e fixados em formol para avaliação histológica e contagem de células caliciformes. Das 110 amostras foram obtidos 48 isolamentos característicos de Brachyspira (43,6%) e destes na qPCR 13 identificadas como B. hyodysenteriae (11,8%) e 5 sendo todas da mesma granja (4,5%) como B. intermedia, 2 amostras foram positivas para ambos os agentes (1,8%) e 28 não foram caracterizadas através da qPCR (25,5%). Não foram observadas alterações histopatológicas no ceco e diferença estatística significativa na contagem de células caliciformes das aves positivas. Conclui-se que a Brachyspira sp. é frequente em granjas de poedeiras e matrizes de corte no Brasil, com especial relevância para a B. intermedia que possui potência patogênico para estas aves.(AU)

Hibridização in situ fluorescente para diagnóstico de Brachyspira hyodysenteriae e B. pilosicoli ­em suínos / Fluorescence in situ hybridization for diagnosis of Brachyspira hyodysenteriae e B. pilosicoli in swine

Disenteria Suína e Colite Espiroquetal são duas enfermidades importantes em suínos causados pela Brachyspira hyodysenteriae e Brachyspira pilosicoli, respectivamente. O diagnóstico eficaz dessas espécies é extremamente importante para a adoção de estratégias adequadas para o controle. Propõe-se avaliar a técnica de hibridização in situ de fluorescência (FISH) para detecção de B. hyodysenteriae e B. pilosicoli em fragmentos histopatológicos de intestino de suínos e compará-la ao PCR duplex. Foram analisadas amostras de fezes e intestinos de suínos de terminação com histórico de diarreia pelas técnicas de reação em cadeia da polimerase duplex (dPCR), hibridização in situ fluorescente (FISH) para diagnóstico dessas bactérias. Foram utilizadas 34 amostras de intestino de suínos de campo positivos para alguma das duas espécies de Brachyspira sp. nos testes de FISH ou PCR. Das 34 amostras analisadas, foram detectadas 28 (82,35%) positivas na PCR e no FISH. Dentre as 29 amostras positivas para B. hyodysenteriae, 23 (79,3%) foram positivas à PCR e 21 (72,4%) no FISH. Os resultados de FISH e PCR não diferiram estatisticamente entre si. Baseado no fato dessa técnica poder ser realizada em tecidos formolizados, ser prática, rápida e associar a marcação especifica do agente com lesões histológicas, o FISH demonstrou ser mais uma alternativa no diagnóstico de Brachyspira hyodysenteriae e B. pilosicoli.(AU)

Growing and finishing pigs are affected by pathogenic spirochetes of the genus Brachyspira sp., which cause a significant economic impact due to direct and indirect losses. Thus, efficient diagnosis of these species enables better technical intervention to prevent or treat diseases. This study aimed to evaluate the fluorescent in situ hybridization (FISH) for the diagnosis of B. hyodysenteriae and B. pilosicoli in histopathologic fragments of pig's intestine and compare it to the duplex PCR. Thirty-four samples collected from pigs positive for these species in at least one of the tests were used in the study. Out of the 34 analyzed intestine samples, 28 (82.35%) were positive by PCR and FISH. Among the 29 B. hyodysenteriae positive samples, 23 (79.3%) were positive by PCR and 21 (72.4%) by FISH. There was no statistical difference among the detection rate of the used tests. Based on the fact this technique can be performed in formalin fixed tissue samples, it is practical, fast and allows the association of labeling a specific agent with histological lesions, FISH has become an alternative diagnostic method for Brachyspira hyodysenteriae and B. pilosicoli.(AU)

Minimum inhibitory concentration of Brazilian Brachyspira hyodysenteriae strains / Concentração inibitória mínima de cepas Brachyspira hyodysenteriae brasileiras

The objectives of this study were to characterize Brachyspira hyodysenteriae isolates and to evaluate the antimicrobial susceptibility patterns of strains obtained from pigs in Brazil based on the minimal inhibitory concentration test (MIC). The MIC was performed for 22 B. hyodysenteriae isolates obtained from 2011 to 2013 using the following antimicrobial drugs: tylosin, tiamulin, valnemulin, doxycycline, lincomycin and tylvalosin. Outbreaks of swine dysentery were diagnosed based on clinical presentation, bacterial isolation, gross and microscopic lesions, duplex PCR for B. hyodysenteriae and B. pilosicoli and nox gene sequencing. All obtained MIC values were consistently higher or equal to the microbiological cut-off described in the literature. The MIC 90 values for the tested drugs were 8µg/ml for doxycycline, >4µg/ml for valnemulin, 8µg/ml for tiamulin, 32µg/ml for tylvalosin, >64µg/ml for lincomycin and >128µg/ml for tylosin. These results largely corroborate those reported in the literature. Tiamulin, doxycycline and tylvalosin showed the lowest MIC results. All of the samples subjected to phylogenetic analysis based on the nox gene sequence exhibited similar results, showing 100% identity to B. hyodysenteriae. This is the first study describing the MIC pattern of B. hyodysenteriae isolated in Brazil.(AU)

Os objetivos deste trabalho foram a caracterização de isolados de Brachyspira hyodysenteriae e avaliar os padrões de sensibilidade antimicrobiana de isolados obtidos a partir de suínos no Brasil com base no teste de concentração inibitória mínima (MIC). A MIC foi realizada em 22 isolados de B. hyodysenteriae obtidos entre 2011 a 2013 usando os seguintes antimicrobianos: tilosina, tiamulina, valnemulina, doxiciclina, lincomicina e tilvalosina. Surtos de disenteria suína foram diagnosticados com base na apresentação clínica, isolamento bacteriano, lesões macroscópicas e microscópicas, PCR duplex para B. hyodysenteriae e B. pilosicoli e sequenciamento do gene nox. Todos os valores de MIC obtidos foram consistentemente mais elevados ou igual ao ponto de corte microbiológica descrito na literatura. Os valores de MIC 90 para os fármacos testados foram de 8 µg / mL para a doxiciclina, > 4 µg/ml de valnemulina, 8 µg / mL para a tiamulina, 32 µg / ml para tilvalosina, > 64 µg / ml para a lincomicina e > 128 µg / ml de tilosina. Estes resultados corroboram em grande parte com os relatados na literatura. Tiamulina, doxiciclina e tilvalosina apresentaram os menores resultados de MIC. Todas as amostras submetidas à análise filogenética com base na sequência do gene nox exibiram resultados semelhantes, indicando 100% de identidade com B. hyodysenteriae. Este é o primeiro estudo que descreve o padrão MIC de B. hyodysenteriae isoladas no Brasil.(AU)

Detección de Brachyspira pilosicoli y otras especies de Brachyspira en granjas avícolas argentinas / Detection of Brachyspira pilosicoli and other Brachyspira species in Argentine poultry farms

Algunas especies del género Brachyspira, como Brachyspira pilosicoli, Brachyspira intermedia y Brachyspira alvinipulli, son especies patógenas capaces de producir enfermedad en gallinas ponedoras. En nuestro país, la presencia de B. pilosicoli y otras especies de Brachyspira ha sido informada en cerdos y en perros, pero no existen antecedentes de su presencia en aves de corral. En este estudio se analizaron muestras de materia fecal y de contenido de ciego de 34 gallinas ponedoras de 4, 12 y 24 meses provenientes de 2 establecimientos por medio del aislamiento, la realización de pruebas bioquímicas y reacción en cadena de la polimerasa. B. pilosicoli y Brachyspira spp. fueron identificadas en muestras tomadas de aves de 12 y 24 meses de edad

Some species of the genus Brachyspira such as Brachyspira pilosicoli, Brachyspira intermedia and Brachyspira alvinipulli are pathogenic species capable of producing disease in laying hens. In our country, the presence of B. pilosicoli and other species of Brachyspira has been reported in pigs and dogs but there is no record of their presence in poultry. Fecal and cecal content samples from 34 laying hens of 4, 12 and 24 months of age from two farms were analyzed by isolation, biochemical tests and PCR. B. pilosicoli and Brachyspira spp. were identified in samples taken from laying hens of 12 and 24 months of age

Comparison of transport media for the isolation and detection of Brachyspira hyodysenteriae / 대한수의학회지

Brachyspira (B.) hyodysenteriae is a causative agent of swine dysentery that is responsible for death and economic losses in the pig industry. It is imperative that clinical samples be delivered fresh for accurate diagnosis. The viability and DNA detection of B. hyodysenteriae using lab-made (phosphate buffered saline and modified tryptic soy broth) or commercial transport media (C, D, and E) were compared by culturing and real-time PCR at 4degrees C or room temperature (RT), respectively. B. hyodysenteriae grown in D (Anaerobe Systems, USA) and E (Starplex Scientific, Canada) media was viable for 4 days at 4degrees C and RT. However, B. hyodysenteriae in A, B, and C (culture swab; BD Biosciences, USA) media were not recovered after 2 days at RT. Ct values for real-time PCR at 4degrees C and RT ranged from 27.2 +/- 2.1 (C) to 29.6 +/- 0.5 (B), and 28.0 +/- 0.9 (E) to 30.2 +/- 1.5 (B), respectively. Considering the field conditions, it is important that transport media is used for specimen isolation and PCR to obtain an accurate diagnosis of swine dysentery.

Prevalence and antimicrobial susceptibility of Brachyspira species in pigs in Korea / 대한수의학회지

The purpose of this study was to investigate the prevalence of Brachyspira species and antimicrobial susceptibility of Brachyspira (B.) hyodysenteriae isolates in Korea. A total of fifty-five Brachyspira species were isolated; five (1.0%) beta-hemolytic Brachyspira species and 50 (10.4%) weak hemolytic Brachyspira species from 116 different diarrheic pig samples and 367 apparently normal pig samples. In farm level, beta hemolytic and weak hemolytic Brachyspira species were detected in 7.4% (5/68) and 19.1% (13/68) of tested pig farms, respectively. By phenotypic and genotypic characterization, all beta hemolytic Brachyspira isolates was classified as group I (B. hyodysenteriae), whereas weak hemolytic Brachyspira species isolates were group III (B. innocens or B. murdochii). B. hyodysenteriae isolates showed high level of minimum inhibition concentrations to macrolide antimicrobials. This study shows that the prevalence of pathogenic B. hyodysenteriae in pigs is low but antimicrobial resistance of the pathogens is high in Korea. This is the first report of the prevalence of Brachyspira group III and antimicrobial susceptibility of B. hyodysenteriae in pigs in Korea. Our results could provide basic data for the management and treatment guidelines of Brachyspira infection.

Pruebas de sensibilidad frente a agentes anti microbianos en cepas de Brachyspira pilosicoli aisladas a partir de ponedoras comerciales en Colombia / Antimicrobial susceptibility test in Brachyspira pilosicoli strains isolated from commercial layers farms in Colombia

Las espiroquetas intestinales del género Brachyspira ocasionan enfermedades importantes en porcinos y aves. Se ha evidenciado un problema de incremento en la presentación de cepas resistentes a los antimicrobianos utilizados normalmente para tratar las espiroquetosis intestinales en porcinos, y esto podría ser aplicable a los aislamientos de aves. Hay muypocos reportes de sensibilidad antimicrobiana in vitro de cepas de Brachyspira spp. aisladas en aves. En este estudio se evaluó la sensibilidad de doce aislamientos de Brachyspira pilosicoli obtenidos de granjas de ponedoras comerciales a los agentes antimicrobianos tiamulina, tilosina y lincomicina, y se estableció la concentración mínima inhibitoria (CMI)mediante la técnica de dilución en agar. Todas las bacterias analizadas fueron sensibles a tiamulina (CMI≤0,1 μg/ml) y lincomicina (CMI 1 μg/ml) y resistentes a tilosina (CMI 5 μg/ml).

Intestinal Spirochaetes of the genus Brachyspira cause important diseases in swine and poultry. An increasing problem in the presentation of resistant strains to the antimicrobial drugs usually used to treat the intestinal spirochaetosis in swine has been evidenced and this could be applicable to the isolations from poultry. There are very few reports of in vitro antimicrobialsusceptibility of Brachyspira spp. isolated from birds. In this study the antimicrobial susceptibility of twelve Brachyspira pilosicoli isolates obtain from commercial layers was evaluated against tiamulin, tylosin and lincomycin establishing the Minimum Inhibitory Concentration (MIC) by agar dilution technique. All bacteria analyzed were sensitive to tiamulin (MIC ≤0,1μg/ml), and lincomycin (MIC 1μg/ml) and resistant to tylosin (MIC 5μg/ml).

The 23S rRNA gene PCR-RFLP used for characterization of porcine intestinal spirochete isolates

Using three reference strains of Brachyspira hyodysenteriae (B204, B234, B169), one B. pilosicoli (P43/6/78), one B. murdochii (56-150), one B. intermedia (PWS/A), one B. innocens (B256) and ten Korean isolates, PCR-RFLP analysis of DNA encoding 23S rRNA was performed to establish a rapid and accurate method for characterizing porcine intestinal spirochetes. Consequently, B. hyodysenteriae and B. pilosicoli revealed different restriction patterns; however, the other three species shared the same pattern. These findings are not consistent with a prior report. Differences in 23S rRNA gene sequences, between two B. murdochii strains, 56-150 and 155-20, were observed. These results indicate that 23S rRNA PCR-RFLP could be used as an identification method for pathogenic Brachyspira spp. (B. hyodysenteriae and B. pilosicoli) as well as an epidemiological tool for characterizing spirochetes isolated from swine.

Simultaneous detection of Lawsonia intracellularis, Brachyspira hyodysenteriae and Salmonella spp. in swine intestinal specimens by multiplex polymerase chain reaction

A multiplex PCR assay was developed for the simultaneous detection of the etiologic agents associated with porcine proliferative enteropathies (PPE), swine dysentery (SD)and porcine salmonellosis (PS)in a single reaction using DNA from swine intestinal samples. Single and multiplex PCR amplification of DNA from Lawsonia intracellularis, Salmonella typhimurium and Brachyspira hyodysenteriae with each primer set produced fragments of the predicted size without any nonspecific amplification, 210-bp, 298-bp and 403-bp bands, respectively. The single PCR assay could detect as little as 100 pg of purified DNA of S. typhimurium and L. intracellularis, and 50 pg of B.hyodysenteriae, respectively. However, multiplex PCR turned out to be 10 times lower sensitivity with S. typhimurium compared with single PCR. With 23 swine intestinal specimens suspected of having PPE, SD and/or PS, the multiplex PCR assay showed identical results with conventional methods except one. In conclusion, this multiplex PCR is a feasible alternative to standard diagnostic methods for detection of L. intracellularis, B. hyodysenteriae and Salmonella spp. from swine intestinal specimens.

Prevalence of Lawsonia intracellularis, Brachyspira hyodysenteriae and Salmonella in swine herds

The prevalence of Lawsonia intracellularis, Brachyspira hyodysenteriae and Salmonella spp. were investigated by multiplex PCR using fecal samples of pigs with diarrhea or a history of diarrhea. The overall herd prevalence of L. intracellularis, B. hyodysenteriae and Salmonella spp. were 46.5%, 37.2% and 51.1%, respectively. Also, the prevalence of L. intracellularis, B. hyodysenteriae and Salmonella spp. among all sampled pigs were 19.9%, 10.8% and 17.7%, respectively. Seventeen of 43 herds were positive with 2 enteric organisms, and 2 herds were positive with L. intracellularis, B. hyodysenteriae and Salmonella spp. simultaneously. It was notable that 11 of 12 herds with more than 2, 000 pigs were affected with Salmonella spp., and that only 2 of 12 the herds were affected with B. hyodysenteriae. This study suggested that herds positive for L. intracellularis, B. hyodysenteriae and Salmonella spp. were distributed throughout Korea, although the relationship among other pathogens such as viral or parasitic ones and/or with metabolic disorders was not determined.

Characterization of Brachyspira hyodysenteriae isolates from Korea

This study was done to characterize diversity in 10 Brachyspira hyodysenteriae isolates in Korea. The isolates were compared with 14 well-characterized non-Korean strains of various Brachyspira species. All Korean isolates showed strong beta haemolysis and had blunt cell ends with 7~14 periplasmic flagella. They produced indole, and did not ferment fructose. They were alpha-glucosidase positive and alpha-galatosidase negative using the APIZYM kit. Using polyclonal antisera raised in rabbits against recognized serotypes, all isolates showed a strong reaction to B. hyodysenteriae antisera E, A and B. Using multilocus enzyme electrophoresis (MLEE) with 15 enzymes and 5 buffer systems, the Korean and non-Korean isolates were divided into 22 electrophoretic types (ETs) and 5 divisions (A, B, C, D and E). Division A corresponded to B. hyodysenteriae, B to B. innocens, C to B. intermedia, D to B. murdochii and E to B. pilosicoli. The 10 Korean isolates of B. hyodysenteriae were relatively diverse, being divided into 9 ETs within MLEE division A. They were all distinct from the non-Korean strains.