Development and preliminary evaluation of a real-time PCR assay for Halioticida noduliformans in abalone tissues

Greeff, Mariska R; Christison, Kevin W.; Macey, Brett M.

doi:10.3354/dao02468

Cited by 5 publications

(4 citation statements)

References 32 publications

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“…Total genomic DNA was extracted from single spore colonies following incubation for 7 days on PDA. A heat lysis DNA extraction protocol was used 27 . Extracted DNA were stored at − 20 °C until needed.…”

Section: Methodsmentioning

confidence: 99%

Fusarium species isolated from post-hatchling loggerhead sea turtles (Caretta caretta) in South Africa

Greeff-Laubscher

Jacobs

2022

Sci Rep

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Species in the Fusarium solani species complex are fast growing, environmental saprophytic fungi. Members of this genus are filamentous fungi with a wide geographical distribution. Fusarium keratoplasticum and F. falciforme have previously been isolated from sea turtle nests and have been associated with high egg mortality rates. Skin lesions were observed in a number of stranded, post-hatchling loggerhead sea turtles (Caretta caretta) in a rehabilitation facility in South Africa. Fungal hyphae were observed in epidermal scrapes of affected turtles and were isolated. The aim of this study was to characterise the Fusarium species that were isolated from these post-hatchling loggerhead sea turtles (Caretta caretta) that washed up on beaches along the South African coastline. Three gene regions were amplified and sequenced, namely the internal transcribed spacer region (ITS), a part of the nuclear large subunit (LSU), and part of the translation elongation factor 1 α (tef1) gene region. Molecular characteristics of strains isolated during this study showed high similarity with Fusarium isolates, which have previously been associated with high egg mortality rates in loggerhead sea turtles. This is the first record of F. keratoplasticum, F. falciforme and F. crassum isolated from stranded post-hatchling loggerhead sea turtles in South Africa.

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Section: Methodsmentioning

confidence: 99%

Fusarium species isolated from post-hatchling loggerhead sea turtles (Caretta caretta) in South Africa

Greeff-Laubscher

Jacobs

2022

Sci Rep

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“…Initial or preliminary detection of Aphanomyces invadans DNA in the fish tissue was achieved through the amplification of a 234 bp fragment of the internal transcribed spacer (ITS) gene region using the A. invadans specific primers recommended by the OIE (OIE, 2019) namely, forward primer Ainvad-2F (5′-TCA TTG TGA GTG AAA CGG TG-3′) and reverse primer Ainvad-ITSR1 (5′-GGC TAA GGT TTC AGT AGT TAG-3′) of Vandersea et al (2006). Duplicate sets of DNA isolated from the tissue sample and the reference samples (following the method of Greeff et al, 2012) were subjected to PCR (Labnet Multigene™ Thermal Cycler (Labnet International, Inc.)). Known A. invadans DNA as well as infected and non-infected reference tissues were included in the analysis to confirm the sensitivity and specificity of the assay.…”

Section: Methodsmentioning

confidence: 99%

Epizootic ulcerative syndrome – First report of evidence from South Africa's largest and premier conservation area, the Kruger National Park

Malherbe

Christison

Wepener

et al. 2019

International Journal for Parasitology: Parasites and Wildlife

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This study reports on the first evidence of genomic material of the causative agent for epizootic ulcerative syndrome (EUS), Aphanomyces invadans, from fish in the Limpopo River system and the Kruger National Park, South Africa. Fourteen fish species were collected from various depressions in the floodplains of the Limpopo and Luvuvhu Rivers in the Makuleke Wetlands during 2015 and 2017. A single individual of Clarias gariepinus was found to have a suspected epizootic ulcerative syndrome (EUS) lesion. Samples were collected and evidence of A. invadans DNA in the samples was found through PCR and amplicon sequencing. The spread of EUS into this premier conservation area is of concern as it could potentially spread across borders and into other naïve river systems with important conservation statuses.

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“…Total genomic DNA was extracted from all single-spore colonies following incubation for 7 d on PDA (Greeff et al 2012). DNA quality and concentrations were confirmed using a Jenway Genova NanoDrop spectrophotometer.…”

Section: Genomic Dna Extraction and Sequencingmentioning

confidence: 99%

First Record of the Water Mold Achlya bisexualis (Saprolegniaceae) Isolated from Ornamental Fish in South Africa

2019

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The order Saprolegniales (Class Oomycota) is a group of fungus-like eukaryotic microorganisms that have been associated with infections in fish and fish eggs. Infections with microorganisms from this order are clearly evident because they have a cotton wool-like appearance. The aim of this study was to characterize and identify an oomycete that was isolated from the eye of an Orange Blotched Peacock Cichlid Aulonacara sp. A sample of cotton wool-like mycelia was isolated and single-spore isolations were conducted. Molecular characterization and phylogenetic analysis of the ITS1-5.8-ITS2 rDNA region for all isolates were used for species identification. Following molecular identification, one isolate was used to culture and characterize the reproductive structures. Physiological characterization entailed incubating the isolate on potato dextrose agar (PDA) at five different temperatures, ranging from 5°C to 25°C, to monitor growth rates. A multiple sequence alignment showed 100% similarity between all of the single-spore isolates and alignment with other Achlya bisexualis strains. Long, coarse hyphae with zoosporangia and gemmae typical of the order Saprolegniales were observed with an optimal growth rate at 25°C. The oomycete that was isolated from an Orange Blotched Peacock Cichlid was identified as A. bisexualis, the first record of this species in South Africa.

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Development and preliminary evaluation of a real-time PCR assay for Halioticida noduliformans in abalone tissues

Cited by 5 publications

References 32 publications

Fusarium species isolated from post-hatchling loggerhead sea turtles (Caretta caretta) in South Africa

Fusarium species isolated from post-hatchling loggerhead sea turtles (Caretta caretta) in South Africa

Epizootic ulcerative syndrome – First report of evidence from South Africa's largest and premier conservation area, the Kruger National Park

First Record of the Water Mold Achlya bisexualis (Saprolegniaceae) Isolated from Ornamental Fish in South Africa

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