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Microbial communities associated with healthy and White syndrome-affected Echinopora lamellosa in aquaria and experimental treatment with the antibiotic ampicillin.

Smith D, Leary P, Craggs J, Bythell J, Sweet M - PLoS ONE (2015)

Bottom Line: A significant difference (R = 0.907, p < 0.001) in 16S rRNA prokaryotic diversity was found between healthy (H), sloughed tissue (ST), WS-affected (WSU) and antibiotic treated (WST) samples.In contrast to previous studies bacterial abundance did not vary significantly (ANOVA, F2, 6 = 1.000, p = 0.422) between H, ST, WSU or WST.Antimicrobial activity (assessed on Vibrio harveyi cultures) was limited in both H and WSU samples (8.1% ±8.2 and 8.0% ±2.5, respectively) and did not differ significantly (Kruskal-Wallis, χ2 (2) = 3.842, p = 0.146).

View Article: PubMed Central - PubMed

Affiliation: School of Biology, Newcastle University, Newcastle upon Tyne, NE1 7RU, United Kingdom; School of Biological Sciences, Medical Biology Centre, Queen's University Belfast, Belfast, BT9 7BL, United Kingdom.

ABSTRACT
Prokaryotic and ciliate communities of healthy and aquarium White Syndrome (WS)-affected coral fragments were screened using denaturing gradient gel electrophoresis (DGGE). A significant difference (R = 0.907, p < 0.001) in 16S rRNA prokaryotic diversity was found between healthy (H), sloughed tissue (ST), WS-affected (WSU) and antibiotic treated (WST) samples. Although 3 Vibrio spp were found in WS-affected samples, two of these species were eliminated following ampicillin treatment, yet lesions continued to advance, suggesting they play a minor or secondary role in the pathogenesis. The third Vibrio sp increased slightly in relative abundance in diseased samples and was abundant in non-diseased samples. Interestingly, a Tenacibaculum sp showed the greatest increase in relative abundance between healthy and WS-affected samples, demonstrating consistently high abundance across all WS-affected and treated samples, suggesting Tenacibaculum sp could be a more likely candidate for pathogenesis in this instance. In contrast to previous studies bacterial abundance did not vary significantly (ANOVA, F2, 6 = 1.000, p = 0.422) between H, ST, WSU or WST. Antimicrobial activity (assessed on Vibrio harveyi cultures) was limited in both H and WSU samples (8.1% ±8.2 and 8.0% ±2.5, respectively) and did not differ significantly (Kruskal-Wallis, χ2 (2) = 3.842, p = 0.146). A Philaster sp, a Cohnilembus sp and a Pseudokeronopsis sp. were present in all WS-affected samples, but not in healthy samples. The exact role of ciliates in WS is yet to be determined, but it is proposed that they are at least responsible for the neat lesion boundary observed in the disease.

No MeSH data available.


Related in: MedlinePlus

Neighbour-joining consensus tree of partial 18S rRNA gene sequences of 4 ciliate species found in Echinopora lamellosa samples affected by White Syndrome in this aquarium experiment.Sequence alignment was carried out using MUSCLE and the neighbour-joining tree was constructed in TreeDyn (via phylogeny.fr), using the Tamura genetic distance model [40] with an opalinid protist; Opalina ranarum (AF141970) as the outgroup, as used by Sweet and Bythell [18]
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pone.0121780.g006: Neighbour-joining consensus tree of partial 18S rRNA gene sequences of 4 ciliate species found in Echinopora lamellosa samples affected by White Syndrome in this aquarium experiment.Sequence alignment was carried out using MUSCLE and the neighbour-joining tree was constructed in TreeDyn (via phylogeny.fr), using the Tamura genetic distance model [40] with an opalinid protist; Opalina ranarum (AF141970) as the outgroup, as used by Sweet and Bythell [18]

Mentions: The ciliate 18S rRNA DGGE profile (Fig. 4) demonstrated the presence of several ciliate species in all white syndrome-affected E. lamellosa samples (treated and untreated), with no ciliates found in healthy samples, indicating the unique association of ciliates with WS pathology. Ciliates appear to be unaffected by treatment with ampicillin, as the ciliate community 18SrRNA gene profile appears relatively unchanged across all ciliate-infected samples (Fig. 4). Ciliate ribotypes present in diseased samples included those most closely related to: Cohnilembus sp. (C1, GenBank accession number Z22878), Loxophyllum sp. (C2 DQ190465), Philaster sp. (C4, JF831359) and Pseudokeronopsis sp. (C3, AY881633). Ciliates C1 (97% similarity to Cohnilembus verminus) and C3 (99% similarity to Pseudokeronopsis sp) appear to be present across all WS-affected samples (treated and untreated) whilst there is more ambiguity over the presence of C4 across all samples (with definite bands only appearing in one WSU and one WST sample) on the DGGE profile (Fig. 4). A PCR product check using Philaster sp. specific primers confirmed the presence of C4 in all ST, WSU and WST samples, with an absence in all H samples (Fig. 5). C4 was more closely related to the more ciliates associated with WS than with Brown Band Disease (Fig. 5). Moreover, C4 was found to be most closely related to Philaster digitformis Morph 3 (Fig. 6), a ciliate isolated from aquarium WS by Sweet, Craggs [29]. C4 differed from Morph 3 by 1 base pair over 445 bases, thus C4 is proposed to be a new morph of P. digitformis, designated as Morph 4. Morph 4 differs from the wild-type WS Morph 1 [18] by 2 base pairs over 445 bases. C2 (98% similarity with Loxophyllum rostratum) is only represented in two WSU and one WST sample on the ciliate 18S rRNA DGGE profile (Fig. 4). A lack of consistency in this latter species indicates that it is unlikely to be involved in the aetiology of WS.


Microbial communities associated with healthy and White syndrome-affected Echinopora lamellosa in aquaria and experimental treatment with the antibiotic ampicillin.

Smith D, Leary P, Craggs J, Bythell J, Sweet M - PLoS ONE (2015)

Neighbour-joining consensus tree of partial 18S rRNA gene sequences of 4 ciliate species found in Echinopora lamellosa samples affected by White Syndrome in this aquarium experiment.Sequence alignment was carried out using MUSCLE and the neighbour-joining tree was constructed in TreeDyn (via phylogeny.fr), using the Tamura genetic distance model [40] with an opalinid protist; Opalina ranarum (AF141970) as the outgroup, as used by Sweet and Bythell [18]
© Copyright Policy
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4368680&req=5

pone.0121780.g006: Neighbour-joining consensus tree of partial 18S rRNA gene sequences of 4 ciliate species found in Echinopora lamellosa samples affected by White Syndrome in this aquarium experiment.Sequence alignment was carried out using MUSCLE and the neighbour-joining tree was constructed in TreeDyn (via phylogeny.fr), using the Tamura genetic distance model [40] with an opalinid protist; Opalina ranarum (AF141970) as the outgroup, as used by Sweet and Bythell [18]
Mentions: The ciliate 18S rRNA DGGE profile (Fig. 4) demonstrated the presence of several ciliate species in all white syndrome-affected E. lamellosa samples (treated and untreated), with no ciliates found in healthy samples, indicating the unique association of ciliates with WS pathology. Ciliates appear to be unaffected by treatment with ampicillin, as the ciliate community 18SrRNA gene profile appears relatively unchanged across all ciliate-infected samples (Fig. 4). Ciliate ribotypes present in diseased samples included those most closely related to: Cohnilembus sp. (C1, GenBank accession number Z22878), Loxophyllum sp. (C2 DQ190465), Philaster sp. (C4, JF831359) and Pseudokeronopsis sp. (C3, AY881633). Ciliates C1 (97% similarity to Cohnilembus verminus) and C3 (99% similarity to Pseudokeronopsis sp) appear to be present across all WS-affected samples (treated and untreated) whilst there is more ambiguity over the presence of C4 across all samples (with definite bands only appearing in one WSU and one WST sample) on the DGGE profile (Fig. 4). A PCR product check using Philaster sp. specific primers confirmed the presence of C4 in all ST, WSU and WST samples, with an absence in all H samples (Fig. 5). C4 was more closely related to the more ciliates associated with WS than with Brown Band Disease (Fig. 5). Moreover, C4 was found to be most closely related to Philaster digitformis Morph 3 (Fig. 6), a ciliate isolated from aquarium WS by Sweet, Craggs [29]. C4 differed from Morph 3 by 1 base pair over 445 bases, thus C4 is proposed to be a new morph of P. digitformis, designated as Morph 4. Morph 4 differs from the wild-type WS Morph 1 [18] by 2 base pairs over 445 bases. C2 (98% similarity with Loxophyllum rostratum) is only represented in two WSU and one WST sample on the ciliate 18S rRNA DGGE profile (Fig. 4). A lack of consistency in this latter species indicates that it is unlikely to be involved in the aetiology of WS.

Bottom Line: A significant difference (R = 0.907, p < 0.001) in 16S rRNA prokaryotic diversity was found between healthy (H), sloughed tissue (ST), WS-affected (WSU) and antibiotic treated (WST) samples.In contrast to previous studies bacterial abundance did not vary significantly (ANOVA, F2, 6 = 1.000, p = 0.422) between H, ST, WSU or WST.Antimicrobial activity (assessed on Vibrio harveyi cultures) was limited in both H and WSU samples (8.1% ±8.2 and 8.0% ±2.5, respectively) and did not differ significantly (Kruskal-Wallis, χ2 (2) = 3.842, p = 0.146).

View Article: PubMed Central - PubMed

Affiliation: School of Biology, Newcastle University, Newcastle upon Tyne, NE1 7RU, United Kingdom; School of Biological Sciences, Medical Biology Centre, Queen's University Belfast, Belfast, BT9 7BL, United Kingdom.

ABSTRACT
Prokaryotic and ciliate communities of healthy and aquarium White Syndrome (WS)-affected coral fragments were screened using denaturing gradient gel electrophoresis (DGGE). A significant difference (R = 0.907, p < 0.001) in 16S rRNA prokaryotic diversity was found between healthy (H), sloughed tissue (ST), WS-affected (WSU) and antibiotic treated (WST) samples. Although 3 Vibrio spp were found in WS-affected samples, two of these species were eliminated following ampicillin treatment, yet lesions continued to advance, suggesting they play a minor or secondary role in the pathogenesis. The third Vibrio sp increased slightly in relative abundance in diseased samples and was abundant in non-diseased samples. Interestingly, a Tenacibaculum sp showed the greatest increase in relative abundance between healthy and WS-affected samples, demonstrating consistently high abundance across all WS-affected and treated samples, suggesting Tenacibaculum sp could be a more likely candidate for pathogenesis in this instance. In contrast to previous studies bacterial abundance did not vary significantly (ANOVA, F2, 6 = 1.000, p = 0.422) between H, ST, WSU or WST. Antimicrobial activity (assessed on Vibrio harveyi cultures) was limited in both H and WSU samples (8.1% ±8.2 and 8.0% ±2.5, respectively) and did not differ significantly (Kruskal-Wallis, χ2 (2) = 3.842, p = 0.146). A Philaster sp, a Cohnilembus sp and a Pseudokeronopsis sp. were present in all WS-affected samples, but not in healthy samples. The exact role of ciliates in WS is yet to be determined, but it is proposed that they are at least responsible for the neat lesion boundary observed in the disease.

No MeSH data available.


Related in: MedlinePlus