The life history patterns of the polychaete, Terebrasabella heterouncinata, a pest of cultured abalone
- Authors: Simon, Carol Anne
- Date: 2005
- Subjects: Polychaeta -- Physiology Sabellidae -- Physiology Abalones -- Diseases Polychaeta -- Reproduction Sabellidae -- Reproduction Abalone culture -- South Africa
- Language: English
- Type: Thesis , Doctoral , PhD
- Identifier: vital:5336 , http://hdl.handle.net/10962/d1005632
- Description: Terebrasabella heterouncinata is a small K-selected sabellid polychaete. It is a simultaneous hermaphrodite with a semi-continuous mode of reproduction, producing relatively few large eggs that are brooded within the parental burrow until the larvae emerge, to settle on the growing edge of the abalone shell. Despite its low fecundity, this worm has become problematic on abalone farms in South Africa. The present study was conducted to gain an understanding of the life history patterns of T. helerouncinata to determine how they contributed to the success of these worms under altered conditions. This study demonstrated that conditions prevalent on abalone farms were conducive to enhancing the reproductive success of this worm, and suggests that larger, more fecund worms may have been selected for in the decade that these worms have been present on the farms. Increased nutrient availability, and possibly the increased stability of the farm environment relative to its natural environment, has led to a 1.5-fold increase in the average size of the worms. Body size was found to be positively correlated with brood size, and this resulted in worms on farms brooding 3 to 4.5 times more offspring at a time than worms from wild abalone. The ability to increase the number of eggs produced at a time may have been limited by the fact that these worms have only two ovaries. Thus, the increase in fecundity may have been related primarily to the increase in the rate at which the eggs were laid by the worms on the farms, and the increase in the coelomic space available for the storage of these rapidly developing eggs. The ability to increase the rate at which oocytes develop may be related to the vitellogenic mechanisms employed by these worms. Vitellogenic oocytes are able to incorporate high molecular weight yolk precursors from the surrounding coelomic fluid through endocytotic activity. This may allow the oocytes to increase the rate at which they incorporate yolk material under conditions of nutrient enrichment. The increase in fecundity did not occur at the expense of offspring size and, presumably, quality. The increased reproductive output on the farms was compounded by a proportionate increase in the number of reproducing worms within the population. In addition, these worms are long-lived (worms from farmed abalone reached a maximum age of approximately 40 months) and exhibit negligible senescence. Thus, their reproductive output did not change significantly with an increase in age. Furthermore, the proportion of the reproductive worms did not decrease with an increase in age. Thus, within the age range tested, worms of all ages have the potential to make equal contributions to population growth. While diet and abalone stocking density could not be identified as having a significant effect on reproductive output and infestation rate under intensive culture conditions, it was demonstrated that in a naïve abalone population, the total intensity of infestation increased exponentially with time. This increase may be a consequence of an increase in fertilisation success. These worms continuously produce entaquasperm that are released into the water column. The sperm are collected by other individuals that then store the sperm in a single spermatheca. The ability to store sperm relieves individuals of a dependence on the synchronisation of spawning of eggs and sperm. As the population size and density increases, there could be more individuals releasing sperm into the water column, resulting in a continuous supply of sperm. The increased production of eggs would therefore not be constrained by a lack of sperm. The stored sperm are released into the brood chamber to fertilise eggs as they are laid, and this would probably increase the fertilisation success in the species. This study also provides evidence to suggest that reproduction in this worm has a seasonal component. Future studies should concentrate on measuring fertilisation success in greater detail, measuring the effect of season on reproduction, determining whether there are genetic differences between worms on farmed and wild abalone and determining whether wild worms have similar life-spans and age-related fecundity as worms on farms.
- Full Text:
- Authors: Simon, Carol Anne
- Date: 2005
- Subjects: Polychaeta -- Physiology Sabellidae -- Physiology Abalones -- Diseases Polychaeta -- Reproduction Sabellidae -- Reproduction Abalone culture -- South Africa
- Language: English
- Type: Thesis , Doctoral , PhD
- Identifier: vital:5336 , http://hdl.handle.net/10962/d1005632
- Description: Terebrasabella heterouncinata is a small K-selected sabellid polychaete. It is a simultaneous hermaphrodite with a semi-continuous mode of reproduction, producing relatively few large eggs that are brooded within the parental burrow until the larvae emerge, to settle on the growing edge of the abalone shell. Despite its low fecundity, this worm has become problematic on abalone farms in South Africa. The present study was conducted to gain an understanding of the life history patterns of T. helerouncinata to determine how they contributed to the success of these worms under altered conditions. This study demonstrated that conditions prevalent on abalone farms were conducive to enhancing the reproductive success of this worm, and suggests that larger, more fecund worms may have been selected for in the decade that these worms have been present on the farms. Increased nutrient availability, and possibly the increased stability of the farm environment relative to its natural environment, has led to a 1.5-fold increase in the average size of the worms. Body size was found to be positively correlated with brood size, and this resulted in worms on farms brooding 3 to 4.5 times more offspring at a time than worms from wild abalone. The ability to increase the number of eggs produced at a time may have been limited by the fact that these worms have only two ovaries. Thus, the increase in fecundity may have been related primarily to the increase in the rate at which the eggs were laid by the worms on the farms, and the increase in the coelomic space available for the storage of these rapidly developing eggs. The ability to increase the rate at which oocytes develop may be related to the vitellogenic mechanisms employed by these worms. Vitellogenic oocytes are able to incorporate high molecular weight yolk precursors from the surrounding coelomic fluid through endocytotic activity. This may allow the oocytes to increase the rate at which they incorporate yolk material under conditions of nutrient enrichment. The increase in fecundity did not occur at the expense of offspring size and, presumably, quality. The increased reproductive output on the farms was compounded by a proportionate increase in the number of reproducing worms within the population. In addition, these worms are long-lived (worms from farmed abalone reached a maximum age of approximately 40 months) and exhibit negligible senescence. Thus, their reproductive output did not change significantly with an increase in age. Furthermore, the proportion of the reproductive worms did not decrease with an increase in age. Thus, within the age range tested, worms of all ages have the potential to make equal contributions to population growth. While diet and abalone stocking density could not be identified as having a significant effect on reproductive output and infestation rate under intensive culture conditions, it was demonstrated that in a naïve abalone population, the total intensity of infestation increased exponentially with time. This increase may be a consequence of an increase in fertilisation success. These worms continuously produce entaquasperm that are released into the water column. The sperm are collected by other individuals that then store the sperm in a single spermatheca. The ability to store sperm relieves individuals of a dependence on the synchronisation of spawning of eggs and sperm. As the population size and density increases, there could be more individuals releasing sperm into the water column, resulting in a continuous supply of sperm. The increased production of eggs would therefore not be constrained by a lack of sperm. The stored sperm are released into the brood chamber to fertilise eggs as they are laid, and this would probably increase the fertilisation success in the species. This study also provides evidence to suggest that reproduction in this worm has a seasonal component. Future studies should concentrate on measuring fertilisation success in greater detail, measuring the effect of season on reproduction, determining whether there are genetic differences between worms on farmed and wild abalone and determining whether wild worms have similar life-spans and age-related fecundity as worms on farms.
- Full Text:
Towards the development of a protocol for the selection of probiotics in marine fish larviculture
- Authors: Vine, Niall Gordon
- Date: 2005
- Subjects: Marine fishes -- Larvae Bacteria Fishes -- Microbiology Fish culture
- Language: English
- Type: Thesis , Doctoral , PhD
- Identifier: vital:5329 , http://hdl.handle.net/10962/d1005175
- Description: Manipulation or control of the microbiological aquaculture environment has been identified as an important focus area for future hatchery development. Subsequently, alternatives to obtain control of the microbiological environment are being sought of which the field of probiotics appears highly promising. Probiotics are usually selected based on various in vitro characteristics, however, the methods used differ and are sometimes unsuccessful due to poor experimentation. The aim of this work is to contribute towards the development of a protocol for the in vitro screening of bacterial candidate probiotics for marine fish larviculture. To reduce the number of candidate probiotics to be tested in vivo, various in vitro experiments need to be conducted, each screening for a particular mode of action – antagonism towards pathogen through production of antimicrobial compounds, growth and attachment to fish intestinal mucus, and the production of other beneficial compounds such as vitamins, fatty-acids and digestive enzymes. A total of 108 bacteria species were isolated from the digestive tract of the adult common clownfish, Amphiprion percula to screen for potential probiotics to be used in clownfish larval rearing. The antagonistic compounds assay identified twelve isolates which showed antagonism towards two or more aquatic pathogens. This was followed by an in vitro test that involved growing the organisms in fish intestinal mucus and modeling their growth parameters. A ranking index (RI) was developed using the lag period (λ) and doubling time (td) of the organism, where (1over λ x td) x 100. Five candidate probionts (AP1-AP5) with varied growth parameters were used for further in vitro experiments. The attachment to mucus assay introduced a novel tool for quantifying competition for attachment sites between candidate probionts and pathogens on mucus. Candidate probiont Pseudoalteromonas AP5 reduced the attachment ability of Vibrio alginolyticus when added before the pathogen and partially out-competed the pathogen for attachment sites when added second. In vitro screening for the production of beneficial compounds tested the candidate probiotics’ ability to produce digestive enzymes - trypsin, lipase and alkaline phosphatase as well as carotenoids and vitamin C. Candidate probiont Pseudoalteromonas AP5 produced high levels of the enzymes (98.2, 34.1 and 91.3 mU product liberated.ml⁻¹, respectively) and contained carotenoids while Kocuria AP4 contained carotenoids but produced low quantities of enzymes (7.8, 0 and 59 mU product liberated.ml⁻¹, respectively). None of the candidate probiotics produced vitamin C. To eliminate potential pathogenic or toxic candidate probionts, Artemia nauplii were exposed to each candidate probiont and the percentage Artemia mortality after 24-hours was determined. Candidate probiont AP2 caused high mortality of Artemia nauplii (98.4%) and was excluded from further studies. Identification of candidate probionts AP3-AP5 was performed using 16S-rDNA molecular techniques and the bacteria were assigned the names Bacillus AP3, Kocuria AP4 and Pseudoalteromonas AP5, respectively. Two methods of larval probiont delivery were tested – attachment to Artemia, and in-water delivery. Attachment to Artemia was high for both Kocuria AP4 and Pseudoalteromonas AP5 (7.2 x10³ and 2.7x10⁴ bacteria.nauplius⁻¹, respectively) while the in-water viability experiment showed that Kocuria AP4 comprised 23.9% of the total culturable water microflora after 24 hours while Pseudoalteromonas AP5 contributed 100%. To validate the findings from the in vitro experiments, in vivo trials using clownfish larvae were performed. Of the four candidate probiotics tested, only Kocuria AP4 showed potential to increase larval survival. In vitro tests produced a better understanding of the possible mode of action and strategies of competition between bacteria, however, the number of criteria in which a candidate probiont is successful in vitro may not be the best predictor for its effectiveness in vivo. Commercial studies that reduce between-treatment variation are required to test predictions about the most suitable probiont or combinations thereof.
- Full Text:
- Authors: Vine, Niall Gordon
- Date: 2005
- Subjects: Marine fishes -- Larvae Bacteria Fishes -- Microbiology Fish culture
- Language: English
- Type: Thesis , Doctoral , PhD
- Identifier: vital:5329 , http://hdl.handle.net/10962/d1005175
- Description: Manipulation or control of the microbiological aquaculture environment has been identified as an important focus area for future hatchery development. Subsequently, alternatives to obtain control of the microbiological environment are being sought of which the field of probiotics appears highly promising. Probiotics are usually selected based on various in vitro characteristics, however, the methods used differ and are sometimes unsuccessful due to poor experimentation. The aim of this work is to contribute towards the development of a protocol for the in vitro screening of bacterial candidate probiotics for marine fish larviculture. To reduce the number of candidate probiotics to be tested in vivo, various in vitro experiments need to be conducted, each screening for a particular mode of action – antagonism towards pathogen through production of antimicrobial compounds, growth and attachment to fish intestinal mucus, and the production of other beneficial compounds such as vitamins, fatty-acids and digestive enzymes. A total of 108 bacteria species were isolated from the digestive tract of the adult common clownfish, Amphiprion percula to screen for potential probiotics to be used in clownfish larval rearing. The antagonistic compounds assay identified twelve isolates which showed antagonism towards two or more aquatic pathogens. This was followed by an in vitro test that involved growing the organisms in fish intestinal mucus and modeling their growth parameters. A ranking index (RI) was developed using the lag period (λ) and doubling time (td) of the organism, where (1over λ x td) x 100. Five candidate probionts (AP1-AP5) with varied growth parameters were used for further in vitro experiments. The attachment to mucus assay introduced a novel tool for quantifying competition for attachment sites between candidate probionts and pathogens on mucus. Candidate probiont Pseudoalteromonas AP5 reduced the attachment ability of Vibrio alginolyticus when added before the pathogen and partially out-competed the pathogen for attachment sites when added second. In vitro screening for the production of beneficial compounds tested the candidate probiotics’ ability to produce digestive enzymes - trypsin, lipase and alkaline phosphatase as well as carotenoids and vitamin C. Candidate probiont Pseudoalteromonas AP5 produced high levels of the enzymes (98.2, 34.1 and 91.3 mU product liberated.ml⁻¹, respectively) and contained carotenoids while Kocuria AP4 contained carotenoids but produced low quantities of enzymes (7.8, 0 and 59 mU product liberated.ml⁻¹, respectively). None of the candidate probiotics produced vitamin C. To eliminate potential pathogenic or toxic candidate probionts, Artemia nauplii were exposed to each candidate probiont and the percentage Artemia mortality after 24-hours was determined. Candidate probiont AP2 caused high mortality of Artemia nauplii (98.4%) and was excluded from further studies. Identification of candidate probionts AP3-AP5 was performed using 16S-rDNA molecular techniques and the bacteria were assigned the names Bacillus AP3, Kocuria AP4 and Pseudoalteromonas AP5, respectively. Two methods of larval probiont delivery were tested – attachment to Artemia, and in-water delivery. Attachment to Artemia was high for both Kocuria AP4 and Pseudoalteromonas AP5 (7.2 x10³ and 2.7x10⁴ bacteria.nauplius⁻¹, respectively) while the in-water viability experiment showed that Kocuria AP4 comprised 23.9% of the total culturable water microflora after 24 hours while Pseudoalteromonas AP5 contributed 100%. To validate the findings from the in vitro experiments, in vivo trials using clownfish larvae were performed. Of the four candidate probiotics tested, only Kocuria AP4 showed potential to increase larval survival. In vitro tests produced a better understanding of the possible mode of action and strategies of competition between bacteria, however, the number of criteria in which a candidate probiont is successful in vitro may not be the best predictor for its effectiveness in vivo. Commercial studies that reduce between-treatment variation are required to test predictions about the most suitable probiont or combinations thereof.
- Full Text:
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