“Cycliplex PCR” confirmation of Fusobacterium necrophorum isolates from captive wallabies: A rapid and accurate approach

Antiabong, John F.; Boardman, Wayne; Smith, Ian; Brown, Melissa H.; Ball, Andrew S.; Goodman, Amanda E.

doi:10.1016/j.anaerobe.2012.12.003

Cited by 10 publications

(3 citation statements)

References 29 publications

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“…Total microbial DNA was extracted from the buccal biofilm and amniotic fluid samples placed in ultrapure water using GenElute Mammalian Genomic DNA Miniprep kit (Sigma-Aldrich, Brazil). Specific primers and amplification conditions were used for the following main microbial groups associated with periodontitis: Fusobacterium nucleatum, Fusobacterium necrophorum, Porphyromonas asaccharolytica, Porphyromonas endodontalis, Porphyromonas gingivalis, Prevotella buccae, Prevotella intermedia, Prevotella melaninogenica, Prevotella nigrescens, Tannerella forsythia, Treponema amylovorum, Treponema denticola, class Mollicutes, and genus Staphylococcaebacteria (Ashimoto et al 1996, Tran et al 1997, Fouad et al 2002, Mayanagi et al 2004, Nadkarni et al 2012, Antiabong et al 2013. Amplifications were performed for 25µL volume containing 1XPCR/Mg 2+ buffer (Boehring Mannheim, Indianapolis/IN, USA), 0.2µL of each deoxynucleoside triphosphate (Pharmacia Biotech, Piscataway/NJ, USA), 0.1µL Taq DNA polymerase (Invitrogen do Brasil, São Paulo/SP, Brazil), 1.0µl of each primer pair (Invitrogen), and 10ng of template.…”

Section: Methodsmentioning

confidence: 99%

Monitoring periodontal lesions and their effects during pregnancy: microbiological aspects of the oral cavity and amniotic fluid in pregnant ewes

et al. 2023

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Periodontitis affects the teeth supporting tissues, leading to tooth loss and damage to animal health. Evidence in humans suggests that oral microorganisms spread systemically, increasing the risk of pregnancy disorders such as miscarriage, prematurity, and low birth weight. This study aimed to verify whether periodontopathogenic microorganisms reach the transplacental unit, culminating in problems in pregnant ewes. After analyzing the oral cavity, 10 clinically healthy pregnant ewes (OGCH group) and 10 pregnant ewes with periodontitis (OGP group) were selected. The subgingival biofilm was collected for the polymerase chain reaction (PCR) test and amniotic fluid for both the PCR and interleukin (IL) analysis. Peripheral blood was collected for complete blood count, and analyses of IL-6, IL1-β, and tumor necrosis factor-α were performed. Placental fragments were collected to assess the inflammatory changes using optical microscopy. After giving birth, both the ewes and their lambs were weighed. On clinical examination, a positive correlation between bleeding and suppuration (correlation index - CI=0.54), suppuration and marginal gingivitis (CI=0.34), and marginal gingivitis and edema (CI=0.54) was observed. The weights of the ewes (p=0.013) and their respective lambs (p=0.04) in the OGP group were lower than those of their OGCH group counterparts. The hematological analysis revealed that the OGP group ewes showed a slight increase in the mean corpuscular volume (p=0.2447), segmented cells (p=0.3375), and eosinophils (p=0.3823) when compared with the OGCH group ewes, without a statistical difference. Regarding the microorganisms detected in the oral cavity, there was a significant difference between the occurrence of periodontal pockets and the presence of Fusobacterium necrophorum (p=0.0328), Porphyromonas asaccharolytica (p=0.0392), and the Mollicutes class (p=0.0352). Staphylococcus genus (p=0.9107) and Archaea domain (p=0.7245) were detected in the amniotic samples of both groups, without a significant difference, whereas P. asaccharolytica (p=0.2685) was only detected in one sample in the OGCH group. The expression of cytokine IL-6 in the OGP group differed significantly between the prepartum and postpartum periods (p=0.0039); moreover, it differed significantly in the postpartum period between the OGCH and OGP groups (p=0.0198). Histological examination showed a higher percentage of placental changes in the OGP group (70%) than in the OGCH group, such as the presence of macrophages, neutrophils, plasma cells, and multifocal areas of calcification. These results do not corroborate the hypothesis of dissemination of oral microorganisms to the placental unit, suggesting that it constitutes placental isolation in sheep.

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Section: Methodsmentioning

confidence: 99%

Monitoring periodontal lesions and their effects during pregnancy: microbiological aspects of the oral cavity and amniotic fluid in pregnant ewes

et al. 2023

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“…6 As Fusobacterium necrophorum is thought to be one of the major etiological agents of periodontal disease in macropods, 11,12 the presence of this organism was investigated in all categories of samples by cycliplex PCR as described previously. 1 Analysis of the stained DGGE gel was carried out by digital scanning, g and an evaluation of the oral microbial community structure associated with healthy, disease, and treatment categories was performed using image analysis software.…”

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A molecular ecological approach to the detection and designation of the etiological agents of a model polymicrobial disease

et al. 2013

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The application of the original Koch postulates and the molecular Koch postulates in the definition of the etiological agents of polymicrobial diseases has received little or no attention. In the present study, denaturing gradient gel electrophoresis (DGGE) of oral samples ( n = 3) from each of 3 categories of animals (healthy, diseased [gingivitis], and then oxytetracycline-treated) was used and revealed different bacterial community structures in a model polymicrobial disease (gingivitis) and after clinical cure. Potential microbes associated with the disease and belonging to the following families were identified: Fusobacteriaceae, Porphyromonadaceae, Flavobacteriaceae, Alcanivoracaceae, Bacteroidaceae, Xanthomonadaceae, and Neisseriaceae. Liquid chromatography–mass spectrophotometric analysis of culturable anaerobic bacteria culture supernatant revealed 3 major compounds (2-hydroxycaproic acid, phenyllactic acid, and indole acetic acid) that differentiated the healthy and disease groups. Results indicate that different microbial community structures were associated with the healthy and disease oral states. The results demonstrate the potential of DGGE as a tool in the detection and designation of etiological agents of polymicrobial diseases.

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The effects of iron limitation and cell density on prokaryotic metabolism and gene expression: Excerpts from Fusobacterium necrophorum strain 774 (sheep isolate)

Antiabong

Ball

Brown

2015

Gene

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“Cycliplex PCR” confirmation of Fusobacterium necrophorum isolates from captive wallabies: A rapid and accurate approach

Cited by 10 publications

References 29 publications

Monitoring periodontal lesions and their effects during pregnancy: microbiological aspects of the oral cavity and amniotic fluid in pregnant ewes

Monitoring periodontal lesions and their effects during pregnancy: microbiological aspects of the oral cavity and amniotic fluid in pregnant ewes

A molecular ecological approach to the detection and designation of the etiological agents of a model polymicrobial disease

The effects of iron limitation and cell density on prokaryotic metabolism and gene expression: Excerpts from Fusobacterium necrophorum strain 774 (sheep isolate)

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