Phenotypic and genotypic characterization of Rhodococcus equi isolated from sputum

Silva, Paulo da; Santos, Adolfo Carlos Barreto; Sato, Daisy Nakamura; Silva, Jaqueline Otero; Medeiros, Marta Inês Cazentini; Carneiro, Ana Maria Machado; Leite, Sérgio Roberto de Andrade; Leite, Clarice Queico Fujimura

doi:10.1016/j.bjid.2012.08.009

Cited by 11 publications

(7 citation statements)

References 26 publications

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“…The human infection with this pathogen was increased since primary isolation (Golub et al, 1967;Silva et al, 2012), the infection with R. equi is thought to be occurring either via inhalation from the soil, ingestion and passing via the alimentary tract or inoculation into a wound or mucous membrane (Prescott, 1991). Exposure to domestic animals or their environment, such as horses and pigs, may play important role in several cases of infection.…”

Section: Discussionmentioning

confidence: 99%

“…A patient infected with R. equi may be treated with unsuitable initial antibiotic due to misdiagnosis with Mycobacterium species, Nocaredias species and diphtheriods. Therefore, early correct diagnosis of the organism and subsequent detection of vapA by PCR must be carried out for accurate therapy and faster recovery of the patient (Silva et al, 2012).…”

Section: Discussionmentioning

confidence: 99%

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Molecular Detection of vapA and vapB Genes in Rhodococcus equi Isolated from Human and Horses by Multiplex PCR in Baghdad

Al-Graibawi¹,

Mohammed²

2015

J. Anim. Health Prod.

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| This study was conducted for Isolation and molecular characterization of Rhodococcus equi (R. equi) isolates from human and horse specimens. A total of 203 samples (100 human sputum and 103 horses' faecal samples) were collected from different places in Baghdad, Iraq during november 2014 to april 2015. The samples were streaked on nalidixic acid-novobiocin-actidione cycloheximide-potassium tellurite (NANAT) agar, then the suspected colonies were subcultured on nutrient and blood agars. Confirmation as R equi was done by assessing the colony on the basis of their morphology and through staining and biochemical tests, followed by testing for the presence of the virulence-associated protein antigen vapA and VapB genes by multiplex PCR. Culture examination of 103 horses' faeces samples revealed the isolation of 7 (6.79%) R. equi, all of them 14.71% (7/49) were obtained from Equestrian club in Al-Ameria location and the bacteriological examination of the 100 human sputum samples showed the recovery of 4 (4.00%) isolates of R equi. The results showed that only 57.14% (4/7) of the R. equi isolates from horses' samples were vapA positive. All the 11 isolates included in this study were negative for vapB gene. This is the first study in Iraq to confirm the presence of vapA gene in R. equi isolated from horse samples indicating the contamination of the horse stables with virulent R. equi. Therefore, it is very important to take care when people deal with the animals or their environment to avoid the infection with this pathogen.

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Section: Discussionmentioning

confidence: 99%

Section: Discussionmentioning

confidence: 99%

Molecular Detection of vapA and vapB Genes in Rhodococcus equi Isolated from Human and Horses by Multiplex PCR in Baghdad

Al-Graibawi¹,

Mohammed²

2015

J. Anim. Health Prod.

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“…The mycolic acids were extracted as described previously [27]. After lipid extraction, mycolic acids were released from cell pellets after incubation in methanol-toluene-sulfuric acid (30:15:1, v/v/v), for 12 h at 75°C.…”

Section: Methodsmentioning

confidence: 99%

Microenvironment ofMycobacterium smegmatisCulture to Induce Cholesterol Consumption Does Cell Wall Remodeling and Enables the Formation of Granuloma-Like Structures

Santos

Marinho

Silva

et al. 2019

BioMed Research International

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Pathogenic species of mycobacteria are known to use the host cholesterol during lung infection as an alternative source of carbon and energy. Mycobacteria culture in minimal medium (MM) has been used as an in vitro experimental model to study the consumption of exogenous cholesterol. Once in MM, different species of mycobacteria start to consume the cholesterol and initiate transcriptional and metabolic adaptations, upregulating the enzymes of the methylcitrate cycle (MCC) and accumulating a variety of primary metabolites that are known to be important substrates for cell wall biosynthesis. We hypothesized that stressful pressure of cultures in MM is able to induce critical adaptation for the bacteria which win the infection. To identify important modifications in the biosynthesis of the cell wall, we cultured the fast-growing and nonpathogenic Mycobacterium smegmatis in MM supplemented with or without glycerol and/or cholesterol. Different from the culture in complete medium Middlebrook 7H9 broth, the bacteria when cultured in MM decreased growth and changed in the accumulation of cell wall molecules. However, the supplementation of MM with glycerol and/or cholesterol recovered the accumulation of phosphatidylinositol mannosides (PIMs) and other phospholipids but maintained growth deceleration. The biosynthesis of lipomannan (LM) and of lipoarabinomannan (LAM) was significantly modulated after culture in MM, independently of glycerol and/or cholesterol supplementation, where LM size was decreased (LM13-25KDa) and LAM increased (LAM37-100KDa), when compared these molecules after bacteria culture in complete medium (LM17-25KDa and LAM37-50KDa). These changes modified the cell surface hydrophobicity and susceptibility against H2O2. The infection of J774 macrophages with M. smegmatis, after culture in MM, induced the formation of granuloma-like structures, while supplementation with cholesterol induced the highest rate of formation of these structures. Taken together, our results identify critical changes in mycobacterial cell wall molecules after culture in MM that induces cholesterol accumulation, helping the mycobacteria to increase their capacity to form granuloma-like structures.

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“…Gram-dodatnia, nieruchliwa, tlenowa, nieregularna pałeczka R. equi, to wewnątrzkomórkowy patogen, który może ulegać transformacji do formy ziarniakowatej [43]. Drobnoustrój bytuje w glebie skażonej odchodami zwierząt roślinożernych, dlatego na zakażenie R. equi przede wszystkim narażone są osoby, które miały kontakt ze skażoną glebą.…”

Section: Rhodococcus Equiunclassified

Usefulness Of Microbial Cytotoxins In The Diagnosis Of Selected Bacterial Infections

Godkowicz

Rudnicka

2021

Postępy Mikrobiologii - Advancements of Microbiology

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Wpłynęło w styczniu, zaakceptowano w czerwcu 2021 r.Streszczenie: Synergistyczna reakcja hemolizy, będąca podstawą testu CAMP znajduje zastosowanie we wstępnej identyfikacji wybranych patogenów bakteryjnych m.in. L. monocytogenes, C. perfringens oraz S. agalactiae. W części eksperymentalnej określono wpływ zastosowanych erytrocytów, czasu inkubacji, odległości między posiewami oraz zróżnicowania użytych szczepów na wystąpienie reakcji CAMP w klasycznym i odwróconym teście CAMP oraz zmodyfikowanej metodzie stosowanej w diagnostyce pałeczek Listeria sp. Wykazano, że optymalnym podłożem do wykonania klasycznego testu CAMP jest podłoże agarowe z 10% dodatkiem odwłóknionej krwi ludzkiej. Do identyfikacji gatunków Listeria sp. najodpowiedniejszym podłożem jest pożywka agarowa z 5% dodatkiem krwi ludzkiej. Podczas gdy erytrocyty owcze stanowiły najlepszy substrat dla hemolizyn C. perfringens, a optymalna odległość posiewu oraz czas inkubacji dla badanych gatunków bakterii wynosiły odpowiednio 1-4 mm oraz 24 h. Przeprowadzając wstępną identyfikacje patogennych drobnoustrojów przy pomocy testu CAMP powinno wykorzystywać się różne typy erytrocytów, ponieważ intensywność reakcji synergistycznej jest różna, zarówno w zależności od szczepu jak i gatunku drobnoustroju.

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Phenotypic and genotypic characterization of Rhodococcus equi isolated from sputum

Abstract: The phenotypic and molecular techniques used constitute a good methodological tool to identify R. equi.

Cited by 11 publications

References 26 publications

Molecular Detection of vapA and vapB Genes in Rhodococcus equi Isolated from Human and Horses by Multiplex PCR in Baghdad

Molecular Detection of vapA and vapB Genes in Rhodococcus equi Isolated from Human and Horses by Multiplex PCR in Baghdad

Microenvironment ofMycobacterium smegmatisCulture to Induce Cholesterol Consumption Does Cell Wall Remodeling and Enables the Formation of Granuloma-Like Structures

Usefulness Of Microbial Cytotoxins In The Diagnosis Of Selected Bacterial Infections

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