The cryopreservation potential and ultrastructure of Agulhas sole Austroglossus pectoralis spermatozoa
- Authors: Markovina, Michael Zeljan
- Date: 2008
- Subjects: Spermatozoa , Spermatozoa -- Cryopreservation , Aquaculture , Fishes -- Breeding , Soleidae , Flatfishes , Agulhas Current (South Africa)
- Language: English
- Type: Thesis , Masters , MSc
- Identifier: vital:5251 , http://hdl.handle.net/10962/d1005094 , Spermatozoa , Spermatozoa -- Cryopreservation , Aquaculture , Fishes -- Breeding , Soleidae , Flatfishes , Agulhas Current (South Africa)
- Description: As the estimated market demand for the Agulhas sole Austroglossus pectoralis exceeds the annual catch from trawlers, this species is a potential aquaculture candidate. Broodstock conditioning and gamete preservation is part of research and development aiming at establishing a breeding protocol for a new aquaculture species. Based on a literature review of the morphology of pleuronectiform spermatozoa, this study was designed firstly, to contribute to the field of spermatozoan morphology by describing the ultrastructure of A. pectoralis spermatozoa. This was followed by an experiment to cryopreserve mature spermatozoa to provide baseline data for future studies on this and related species. The testis of A. pectoralis was a paired structure encased in a membrane, the tunica albuginea. The primary testis was located on the dorsal surface of the rib cage and the secondary testis on the ventral side. The testis was of an unrestricted spermatogonial type, based upon observations of spermatogonia along the entire length of the lobule. Mature spermatozoa of A. pectoralis had an acrosome-free ovoid head 1.68 ± 1.6μm in length and 1.7 ± 1.6μm in diameter, a short mid-piece of 0.5 ± 0.1μm in length, containing 7 irregularly shaped mitochondria forming a ring-like structure at the base of the nucleus. The flagellae were 47.4 ± 4.8μm in length, most with two plasma membrane lateral fin-like projections. However, some flagellae had either zero or three lateral fin projections. Cross-sections of the flagellae showed an axenome with a 9+2 microtubule configuration. The proximal and distal centriols were coaxal, situated deep within the nuclear fossa. The structure of A. pectoralis spermatozoa conformed to the type 1 ect-aquasperm, also found in externally fertilizing species. This type has been suggested to be the plesiomorphic form in Neopterigians. Finally, this study contributed to a cryopreservation protocol for A. pectoralis spermatozoa by testing the two cryoprotectants dimethyl sulphoxide (DMSO) and glycerol. Glycerol, at a concentration of 10%, offered better cryoprotection than DMSO. This was established using flow cytometry analysis of post-thaw nuclear membrane integrity after 64 days of storage in liquid nitrogen. The toxicity of DMSO to isolated cellular proteins may have resulted in DMSO-treated sperm having the highest percent (35.2% ± 3.2%) of non-viable cells compared with 23.0% ± 2.5% and 27.8% ± 3.4% for glycerol and the control, respectively. The presence of sucrose in the Modified Mounib Medium extender solution may explain why 45.5% ± 5% of the sperm cells were potentially viable in the control treatment. Initially, the white margined sole Dagatichthys marginatus (Soleidae) was selected as the most suitable candidate for flatfish aquaculture in South Africa. Thus, the aim of this study was to investigate the cryogenic potential and ultrastructure of D. marginatus spermatozoa. However, due to a skewed sex ratio, there were not enough males available to study this species. A skewed sex ratio is common amongst soleids, thus, the need to develop effective cryopreservation methods and to develop an understanding of sperm morphology so that the best time for cryopreservation can be chosen. In conclusion, this first description of spermatozan morphology of A. pectoralis contributed to our understanding of soleid sperm ultrastructure. In addition, a comparison of testis appearance between fish sampled just prior to spawning season and fish with mature sperm provided information on the spawning season of this species. The findings from the cryopreservation experiment suggested that glycerol was a feasible cryoprotectant for this species when sperm was prepared under field conditions.
- Full Text:
- Date Issued: 2008
- Authors: Markovina, Michael Zeljan
- Date: 2008
- Subjects: Spermatozoa , Spermatozoa -- Cryopreservation , Aquaculture , Fishes -- Breeding , Soleidae , Flatfishes , Agulhas Current (South Africa)
- Language: English
- Type: Thesis , Masters , MSc
- Identifier: vital:5251 , http://hdl.handle.net/10962/d1005094 , Spermatozoa , Spermatozoa -- Cryopreservation , Aquaculture , Fishes -- Breeding , Soleidae , Flatfishes , Agulhas Current (South Africa)
- Description: As the estimated market demand for the Agulhas sole Austroglossus pectoralis exceeds the annual catch from trawlers, this species is a potential aquaculture candidate. Broodstock conditioning and gamete preservation is part of research and development aiming at establishing a breeding protocol for a new aquaculture species. Based on a literature review of the morphology of pleuronectiform spermatozoa, this study was designed firstly, to contribute to the field of spermatozoan morphology by describing the ultrastructure of A. pectoralis spermatozoa. This was followed by an experiment to cryopreserve mature spermatozoa to provide baseline data for future studies on this and related species. The testis of A. pectoralis was a paired structure encased in a membrane, the tunica albuginea. The primary testis was located on the dorsal surface of the rib cage and the secondary testis on the ventral side. The testis was of an unrestricted spermatogonial type, based upon observations of spermatogonia along the entire length of the lobule. Mature spermatozoa of A. pectoralis had an acrosome-free ovoid head 1.68 ± 1.6μm in length and 1.7 ± 1.6μm in diameter, a short mid-piece of 0.5 ± 0.1μm in length, containing 7 irregularly shaped mitochondria forming a ring-like structure at the base of the nucleus. The flagellae were 47.4 ± 4.8μm in length, most with two plasma membrane lateral fin-like projections. However, some flagellae had either zero or three lateral fin projections. Cross-sections of the flagellae showed an axenome with a 9+2 microtubule configuration. The proximal and distal centriols were coaxal, situated deep within the nuclear fossa. The structure of A. pectoralis spermatozoa conformed to the type 1 ect-aquasperm, also found in externally fertilizing species. This type has been suggested to be the plesiomorphic form in Neopterigians. Finally, this study contributed to a cryopreservation protocol for A. pectoralis spermatozoa by testing the two cryoprotectants dimethyl sulphoxide (DMSO) and glycerol. Glycerol, at a concentration of 10%, offered better cryoprotection than DMSO. This was established using flow cytometry analysis of post-thaw nuclear membrane integrity after 64 days of storage in liquid nitrogen. The toxicity of DMSO to isolated cellular proteins may have resulted in DMSO-treated sperm having the highest percent (35.2% ± 3.2%) of non-viable cells compared with 23.0% ± 2.5% and 27.8% ± 3.4% for glycerol and the control, respectively. The presence of sucrose in the Modified Mounib Medium extender solution may explain why 45.5% ± 5% of the sperm cells were potentially viable in the control treatment. Initially, the white margined sole Dagatichthys marginatus (Soleidae) was selected as the most suitable candidate for flatfish aquaculture in South Africa. Thus, the aim of this study was to investigate the cryogenic potential and ultrastructure of D. marginatus spermatozoa. However, due to a skewed sex ratio, there were not enough males available to study this species. A skewed sex ratio is common amongst soleids, thus, the need to develop effective cryopreservation methods and to develop an understanding of sperm morphology so that the best time for cryopreservation can be chosen. In conclusion, this first description of spermatozan morphology of A. pectoralis contributed to our understanding of soleid sperm ultrastructure. In addition, a comparison of testis appearance between fish sampled just prior to spawning season and fish with mature sperm provided information on the spawning season of this species. The findings from the cryopreservation experiment suggested that glycerol was a feasible cryoprotectant for this species when sperm was prepared under field conditions.
- Full Text:
- Date Issued: 2008
The potential role of Lake Malawi National Park sanctuary areas for biological control of schistosomiasis and development of a sustainable fishery
- Authors: Msukwa, Amulike Victor
- Date: 1998
- Subjects: Lake Malawi National Park (Malawi) , Schistosomiasis -- Prevention , Mollusks , Aquatic ecology -- Study and teaching , Fishes -- Breeding
- Language: English
- Type: Thesis , Masters , MSc
- Identifier: vital:5313 , http://hdl.handle.net/10962/d1005158 , Lake Malawi National Park (Malawi) , Schistosomiasis -- Prevention , Mollusks , Aquatic ecology -- Study and teaching , Fishes -- Breeding
- Description: The potential role of sanctuary areas for biological control of Schistosomiasis and development of sustainable fisheries was investigated at Cape Maclear, Lake Malawi National Park (LMNP). There has been a recent increase in the incidence of schistosomiasis infection which is a threat to the local community as well as the tourism industry which is the major source of income to LMNP as well as Chembe Village. At the same place there is increasing fishing pressure due to growing human population and declining fish resource. The increase in the incidence of schistosomiasis transmission was attributed in part to over-fishing of the molluscivorous fish which are believed to control the vector snails for schistosomiasis. Four molluscivorous fish species, Trematocranus placodon; Trematocranus microstoma; Mylochromis sphaerodon and Mylochromis anaphyrmus were reported to account for more than 90% of the fishes (by numerical abundance) which feed on the gastropods above 15 metre depth. The gastropod numbers was reported to be highest at 1.5 to 4.5 metre depth. Of the four molluscivores, T. placodon was proposed as a biological control agent for schistosomiasis based upon the previous observations of its feeding habits in artificial conditions. Captive propagation of T. placodon for reintroduction at Cape Maclear in Lake Malawi has been proposed. The present study aimed at providing baseline data required to test the hypotheses that: 1) Over-fishing of the molluscivorous fish has resulted to the increased incidence of schistosomiasis at Cape Maclear. A sub hypothesis to this was that an extension of the LMNP can act as a sanctuary area for the biological control of schistosomiasis by protecting molluscivorous fish which could control schistosomiasis vector snails. 2) A park initially designed to protect the colourful rock dwelling fish and for promotion of tourism may not effectively protect the food fish. To test the first hypothesis, the biology and ecology of T. placodon were investigated with a view to evaluating the effect this species could have on the schistosomiasis vector snail population and hence the control of bilharzia in the lake. The proportions of various gastropod species at Cape Maclear was compared with those found in T. placodon guts. Comparisons of T. placodon abundance and demographic structure inside and outside LMNP were made. To test the second hypothesis, this study investigated the food fish species that use LMNP 100 m protected zone and some basic ecological factors to appreciate the extent to which the adjacent fishery might benefit from their use of the park waters. T. placodon numerical abundance (number of individuals per unit area) ranged from 5.7 to 40.5 /200 m² and it significantly (P< 0.05) varied between sampling sites. Otter Point and Mitande which are inside the park had the lowest abundance as compared to the other three sites; Nguli inside the park; Fisheries and Nchenga outside the park. Two sites in the park, Otter Point and Mitande, had a greater proportion of mature T. placodon individuals than all other sites. The abundance of T. placodon fluctuated significantly from month to month at Nchenga, Nguli and Fisheries (X² test, P<0.0001 for all the three sites) and insignificantly (P>0.05) at Otter Point and Mitande (X² test). T. placodon densities found in the present study corresponded to the peak density of 30 individuals / 200 m² reported in 1986 but did not correspond to that of 1.0 / 200 m² for 1994. There was no evidence to support the previous reports that T. placodon abundance had decreased tremendously from 1986. The reason suggested to account for the discrepancies of T. placodon abundance reported in the present study and other studies was inadequate sampling in the previous studies which did not take into account spatial and temporal variability in T. placodon abundance. The findings reported in this thesis show that there is no need for captive propagation of T. placodon to be reintroduced into the lake at Cape Maclear and that it may prove to be unsuccessful. However, since juvenile T. placodon dominated in abundance at the three sites along the major beach which is outside the park boundaries, it is suggested that the park boundaries be extended to this area so that T. placodon should be protected to allow individuals to grow to bigger size which would be more effective for gastropod control. T. placodon between 60 mm and 80 mm TL fed on benthic insects, phytoplankton and from detritus material. Individuals between 80 mm and 100 mm fed on a mixture of benthic insects, fish scales and small gastropods and at sizes greater than 100 mm individuals specialized feeding on gastropods. Gastropods of five genera were taken and they were: Melanoides , Bulinus, Gabiella, Lanistes and Bellamya. Of these genera Melanoides fonned the greatest part of T. placodon diet. Bulinus was the second most abundant genus but compared to Melanoides its proportion was very small. Of the three Bulinus species taken by T. placodon, B. globosus, is a confirmed vector for Schistosoma haematobium which is prevalent at Cape Maclear. This species was eaten in the least quantities. A comparison ofthe five gastropod proportions in T. placodon diet and in the habitats they occupy showed that Melanoides were taken in proportionately more quantities than Bulinus at most sites. These findings contrasted the previous reports that T. placodon preferred Bulinus to Melanoides. By applying the optimal foraging theory which predicts that an animal species searching for food will go for the type of prey with the highest profitability, it is concluded that the Bulinus cannot be eliminated completely by molluscivores because if their population size falls below a certain level, the fish will switch to other gastropod types. It is concluded that the increase in schistosomiasis may not be necessarily due to overfishing the molluscivorous fish but could be due to the fact that there has been an increase in the proportion of the B. globosus albeit in small numbers which are infected with schistosomiasis parasites. An integrated approach to schistosomiasis control at Cape Maclear comprising vector control, improved water supply, sanitation and health education is suggested since no method can be effective in isolation. Few food fish species were observed to use the park at various times, varying from one species to another with regards to duration, life history stages and abundance. Only a few fish species taken by the adjacent artisanal and commercial fisheries were represented among those observed in the park. This was attributed to the limited diversity of habitat types covered. Only small population size of some species visited the protected area and only part of the life cycle of some species were observed in the park. The use of the park area was seasonal for some species and the protected zone boundaries can be crossed more than once within a day because 100 m distance is just a few minutes swim by fish. Under such circumstances the park cannot function as an effective sanctuary for food fish. An increase of the park size may be a better option to effectively protect the food fish.
- Full Text:
- Date Issued: 1998
- Authors: Msukwa, Amulike Victor
- Date: 1998
- Subjects: Lake Malawi National Park (Malawi) , Schistosomiasis -- Prevention , Mollusks , Aquatic ecology -- Study and teaching , Fishes -- Breeding
- Language: English
- Type: Thesis , Masters , MSc
- Identifier: vital:5313 , http://hdl.handle.net/10962/d1005158 , Lake Malawi National Park (Malawi) , Schistosomiasis -- Prevention , Mollusks , Aquatic ecology -- Study and teaching , Fishes -- Breeding
- Description: The potential role of sanctuary areas for biological control of Schistosomiasis and development of sustainable fisheries was investigated at Cape Maclear, Lake Malawi National Park (LMNP). There has been a recent increase in the incidence of schistosomiasis infection which is a threat to the local community as well as the tourism industry which is the major source of income to LMNP as well as Chembe Village. At the same place there is increasing fishing pressure due to growing human population and declining fish resource. The increase in the incidence of schistosomiasis transmission was attributed in part to over-fishing of the molluscivorous fish which are believed to control the vector snails for schistosomiasis. Four molluscivorous fish species, Trematocranus placodon; Trematocranus microstoma; Mylochromis sphaerodon and Mylochromis anaphyrmus were reported to account for more than 90% of the fishes (by numerical abundance) which feed on the gastropods above 15 metre depth. The gastropod numbers was reported to be highest at 1.5 to 4.5 metre depth. Of the four molluscivores, T. placodon was proposed as a biological control agent for schistosomiasis based upon the previous observations of its feeding habits in artificial conditions. Captive propagation of T. placodon for reintroduction at Cape Maclear in Lake Malawi has been proposed. The present study aimed at providing baseline data required to test the hypotheses that: 1) Over-fishing of the molluscivorous fish has resulted to the increased incidence of schistosomiasis at Cape Maclear. A sub hypothesis to this was that an extension of the LMNP can act as a sanctuary area for the biological control of schistosomiasis by protecting molluscivorous fish which could control schistosomiasis vector snails. 2) A park initially designed to protect the colourful rock dwelling fish and for promotion of tourism may not effectively protect the food fish. To test the first hypothesis, the biology and ecology of T. placodon were investigated with a view to evaluating the effect this species could have on the schistosomiasis vector snail population and hence the control of bilharzia in the lake. The proportions of various gastropod species at Cape Maclear was compared with those found in T. placodon guts. Comparisons of T. placodon abundance and demographic structure inside and outside LMNP were made. To test the second hypothesis, this study investigated the food fish species that use LMNP 100 m protected zone and some basic ecological factors to appreciate the extent to which the adjacent fishery might benefit from their use of the park waters. T. placodon numerical abundance (number of individuals per unit area) ranged from 5.7 to 40.5 /200 m² and it significantly (P< 0.05) varied between sampling sites. Otter Point and Mitande which are inside the park had the lowest abundance as compared to the other three sites; Nguli inside the park; Fisheries and Nchenga outside the park. Two sites in the park, Otter Point and Mitande, had a greater proportion of mature T. placodon individuals than all other sites. The abundance of T. placodon fluctuated significantly from month to month at Nchenga, Nguli and Fisheries (X² test, P<0.0001 for all the three sites) and insignificantly (P>0.05) at Otter Point and Mitande (X² test). T. placodon densities found in the present study corresponded to the peak density of 30 individuals / 200 m² reported in 1986 but did not correspond to that of 1.0 / 200 m² for 1994. There was no evidence to support the previous reports that T. placodon abundance had decreased tremendously from 1986. The reason suggested to account for the discrepancies of T. placodon abundance reported in the present study and other studies was inadequate sampling in the previous studies which did not take into account spatial and temporal variability in T. placodon abundance. The findings reported in this thesis show that there is no need for captive propagation of T. placodon to be reintroduced into the lake at Cape Maclear and that it may prove to be unsuccessful. However, since juvenile T. placodon dominated in abundance at the three sites along the major beach which is outside the park boundaries, it is suggested that the park boundaries be extended to this area so that T. placodon should be protected to allow individuals to grow to bigger size which would be more effective for gastropod control. T. placodon between 60 mm and 80 mm TL fed on benthic insects, phytoplankton and from detritus material. Individuals between 80 mm and 100 mm fed on a mixture of benthic insects, fish scales and small gastropods and at sizes greater than 100 mm individuals specialized feeding on gastropods. Gastropods of five genera were taken and they were: Melanoides , Bulinus, Gabiella, Lanistes and Bellamya. Of these genera Melanoides fonned the greatest part of T. placodon diet. Bulinus was the second most abundant genus but compared to Melanoides its proportion was very small. Of the three Bulinus species taken by T. placodon, B. globosus, is a confirmed vector for Schistosoma haematobium which is prevalent at Cape Maclear. This species was eaten in the least quantities. A comparison ofthe five gastropod proportions in T. placodon diet and in the habitats they occupy showed that Melanoides were taken in proportionately more quantities than Bulinus at most sites. These findings contrasted the previous reports that T. placodon preferred Bulinus to Melanoides. By applying the optimal foraging theory which predicts that an animal species searching for food will go for the type of prey with the highest profitability, it is concluded that the Bulinus cannot be eliminated completely by molluscivores because if their population size falls below a certain level, the fish will switch to other gastropod types. It is concluded that the increase in schistosomiasis may not be necessarily due to overfishing the molluscivorous fish but could be due to the fact that there has been an increase in the proportion of the B. globosus albeit in small numbers which are infected with schistosomiasis parasites. An integrated approach to schistosomiasis control at Cape Maclear comprising vector control, improved water supply, sanitation and health education is suggested since no method can be effective in isolation. Few food fish species were observed to use the park at various times, varying from one species to another with regards to duration, life history stages and abundance. Only a few fish species taken by the adjacent artisanal and commercial fisheries were represented among those observed in the park. This was attributed to the limited diversity of habitat types covered. Only small population size of some species visited the protected area and only part of the life cycle of some species were observed in the park. The use of the park area was seasonal for some species and the protected zone boundaries can be crossed more than once within a day because 100 m distance is just a few minutes swim by fish. Under such circumstances the park cannot function as an effective sanctuary for food fish. An increase of the park size may be a better option to effectively protect the food fish.
- Full Text:
- Date Issued: 1998
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