Displaying publications 1 - 20 of 23 in total

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  1. Preliminary studies on cryopreservation of snakehead (Channa striata) embryos
    Mohd Sharifuddin M, Siti Azizah MN
    Cryobiology, 2014 Aug;69(1):1-9.
    PMID: 24726775 DOI: 10.1016/j.cryobiol.2014.04.001
    This paper reports the findings of the ongoing studies on cryopreservation of the snakehead, Channa striata embryos. The specific objective of this study was to collect data on the sensitivity of C. striata embryo hatching rate to low temperatures at two different developmental stages in the presence of four different cryoprotectants. Embryos at morula and heartbeat stages were selected and incubated in 1M dimethyl sulfoxide (Me2SO), 1M ethylene glycol (EG), 1M methanol (MeOH) and 0.1M sucrose solutions at different temperatures for a period of time. Embryos were kept at 24 °C (control), 15 °C, 4 °C and -2 °C for 5 min, 1h and 3h. Following these treatments, the embryos were then transferred into a 24 °C water bath until hatch to evaluate the hatching rate. The results showed that there was a significant decrease of hatching rate in both developmental stages following exposure to 4 °C and -2 °C at 1h and 3h exposure in each treatment. Heartbeat stage was more tolerant against chilling at -2 °C for 3h exposure in Me2SO followed by MeOH, sucrose and EG. Further studies will be conducted to find the best method to preserve embryos for long term storage.
  2. Isolation of microsatellites in the bighead catfish, Clarias macrocephalus and cross-amplification in selected Clarias species
    Nazia AK, Siti Azizah MN
    Mol Biol Rep, 2014 Mar;41(3):1207-13.
    PMID: 24381108 DOI: 10.1007/s11033-013-2965-9
    The present study documents the isolation of eight polymorphic microsatellite markers from the bighead catfish, Clarias macrocephalus and cross-amplification in two other catfish species. The number of alleles per locus in C. macrocephalus ranged from 2 to 21. The most polymorphic locus was NCm-G12 with 21 alleles while the least polymorphic locus was NCm-H2 with only two alleles. Locus NCm-F8 significantly deviated from Hardy-Weinberg equilibrium (P value <0.05) after Bonferroni correction. Linkage disequilibrium was non-significant in all loci comparisons. The observed and expected heterozygosities varied from 0.033 to 0.967 and from 0.033 to 0.942, respectively. Mean polymorphic information content for the eight loci was 0.765. Cross-amplification was successfully performed with two other catfish species, C. batrachus and C. meladerma for all eight loci. Locus NCm-D8 was monomorphic in both species while NCm-F8 was monomorphic only in C. batrachus. These newly developed markers would be useful for better management and conservation of the economically important C. macrocephalus species.
  3. Fulltext Phylogenetic study and barcoding of the blood cockle, Tegillarca granosa, found on the west coast of peninsular Malaysia using the COI gene
    Chee SY, Devakie MN, Siti Azizah MN
    Genet. Mol. Res., 2011;10(2):1237-44.
    PMID: 21732288 DOI: 10.4238/vol10-2gmr1104
    Blood cockles are among the most economically important brackish water invertebrates found in Malaysia. However, our knowledge of blood cockle phylogeny and systematics is rudimentary, especially for the species Tegillarca granosa. It is unclear, for instance, whether the cockles occurring on the west coast of peninsular Malaysia constitute a single species, or multiple, phylogenetically distinct species. We performed the first DNA molecular phylogenetic analysis of T. granosa to distinguish it from other related species found in other parts of the world and to create a DNA database for the species. An approximately 585-nucleotide fragment of the mitochondrial DNA (cytochrome oxidase I, COI) was sequenced for 150 individual cockles, representing 10 populations: three from the north, four from the central part and three from the southern part of peninsular Malaysia. Phylogenetic analyses of the resulting dataset yielded tree topologies that not only showed the relationship between T. granosa and its closest relatives but its position in the evolutionary tree. Three mitochondrial clades were evident, each containing an individual genus. Using the mutation rate of the COI gene, the divergence time between T. granosa and its closest related species was estimated to be 460 thousand years ago. This study provides a phylogenetic framework for this ecologically prominent and commercially important cockle species.
  4. Fulltext Utilization of molecular markers for the conservation of blood cockles, Anadara granosa (Arcidae)
    Chee SY, Azizah MN, Devakie MN
    Genet. Mol. Res., 2011;10(2):1245-61.
    PMID: 21732289 DOI: 10.4238/vol10-2gmr1103
    We examined genetic variation in blood cockles in an effort to obtain information useful for the sustainability, management, and the stability of this species as a major commodity in the fisheries sector. Ten populations of cockles were sampled from the north to the south of the west coast of peninsular Malaysia. The cockles were collected in collaboration with the Fisheries Research Institute, Penang. The population genetic analysis of the cockles were studied via RAPD-PCR and mtDNA sequencing. Three hundred individuals were analyzed with RAPD-PCR experiments. High gene diversity over all loci was observed (Shannon index = 0.549 ± 0.056 and Nei's gene diversity = 0.4852 ± 0.0430 among 35 loci). The second method, mtDNA sequencing, was employed to complement the information obtained from RAPD-PCR. The gene selected for mtDNA sequencing was cytochrome c oxidase I (COI). One hundred and fifty individuals were sequenced, yielding a partial gene of 585 bp. Statistical analysis showed homogeneity in general but did reveal some degree of variability between the populations in Johor and the rest of the populations. The Mantel test showed a positive but nonsignificant correlation between geographic and genetic distances (r = 0.2710, P = 0.622), as in the RAPD analysis. We propose that the homogeneity between distant populations is caused by two factors: 1) the translocation of the spats; 2) larvae are carried by current movement from the north of the peninsula to the south. The different genetic composition found in Johor could be due to pollution, mutagenic substances or physical factors such as the depth of the water column. This population genetic study is the first for this species in peninsular Malaysia. The data from this study have important implications for fishery management, conservation of blood cockles and translocation policies for aquaculture and stock enhancement programs.
  5. Fulltext Spawning seasons of Rasbora tawarensis (Pisces: Cyprinidae) in Lake Laut Tawar, Aceh province, Indonesia
    Muchlisin ZA, Musman M, Azizah MN
    Reprod Biol Endocrinol, 2010;8:49.
    PMID: 20482777 DOI: 10.1186/1477-7827-8-49
    Rasbora tawarensis is an endemic freshwater fish in Lake Laut Tawar, Aceh Province, Indonesia. Unfortunately, its status is regarded as critical endangered with populations decreasing in recent years. To date no information on the spawning activities of the fish are available. Therefore, this study provides a contribution to the knowledge on reproductive biology of R. tawarensis especially on spawning seasons as well as basic information for conservation of the species.
  6. Genetic variation of Haemonchus contortus (Trichostrongylidae) in sheep and goats from Malaysia and Yemen
    Gharamah AA, Azizah MN, Rahman WA
    Vet Parasitol, 2012 Sep 10;188(3-4):268-76.
    PMID: 22538095 DOI: 10.1016/j.vetpar.2012.04.003
    The large stomach worm, Haemonchus contortus, commonly known as "the barber's pole worm", is a blood-sucking nematode found in the abomasa of sheep and goats. This work is the first documentation on the ND4 sequences of H. contortus from sheep and goats in Malaysia and Yemen and the results provide a preliminary insight on the genetic differences of H. contortus found in the two countries. In general, this study showed a high degree of diversity and low population structure of this species within the same country in comparison with higher genetic structuring at a wider geographical scale. The results also showed that the majority of genetic variance was within H. contortus populations. The Malaysian sheep and goat populations investigated appeared to share the same isolate of H. contortus while different isolates may be found in Yemen which must be taken into account in the design of an effective control strategy. Analysis of the internal transcribed spacer-2 (ITS-2) confirmed that all samples investigated in this study belonged to H. contortus. However presence of other Haemonchus species parasitizing these two hosts can only be confirmed by further detailed studies.
  7. Isolation and multiplex genotyping of polymorphic microsatellite DNA markers in the snakehead murrel, Channa striata
    Jamsari AF, Min-Pau T, Siti-Azizah MN
    Genet Mol Biol, 2011 Apr;34(2):345-7.
    PMID: 21734840
    Seven polymorphic microsatellite loci were isolated and characterized for the snakehead murrel, Channa striata (Channidae), a valuable tropical freshwater fish species. Among 25 specimens collected from Kedah state in Malaysia, the number of alleles per locus ranged from 2 to 7. Observed and expected heterozygosities ranged from 0.120 to 0.880 and 0.117 to 0.698, respectively. A single locus (CS1-C07) was significantly deviated from Hardy-Weinberg equilibrium after Bonferroni correction. These novel markers would be useful for population genetic studies of the C. striata.
  8. Fulltext Remarkably low genetic variation but high population differentiation in the climbing perch, Anabas testudineus (Anabantidae), based on the mtDNA control region
    Jamsari AF, Muchlisin ZA, Musri M, Siti Azizah MN
    Genet. Mol. Res., 2010;9(3):1836-43.
    PMID: 20845309 DOI: 10.4238/vol9-3gmr933
    Anabas testudineus (Anabantidae) is an important food fish in Southeast Asia. We analyzed the mitochondrial DNA control region sequence data to evaluate the genetic variability and population structure of this species. Sixty specimens were collected from four populations in Sumatra and two populations in Peninsular Malaysia. We found a very low level of genetic variability, with five of the six populations exhibiting total absence of genetic variation. Based on analysis of molecular variance, 84.72% of the total variation was among populations and 15.28% within populations. A geographical division based on FST values indicated highly significant genetic differentiation among populations from the four drainage systems: Aceh, Sumatra Utara, Pulau Pinang, and Terengganu (FST ranging from 0.633 to 1.000). No phylogeographic relationships among populations were detected, despite the generation of four distinct clades in a neighbor-joining phylogenetic tree.
  9. Fulltext Phylogeographic pattern of the striped snakehead, Channa striata in Sundaland: ancient river connectivity, geographical and anthropogenic signatures [corrected]
    Tan MP, Jamsari AF, Siti Azizah MN
    PLoS One, 2012;7(12):e52089.
    PMID: 23284881 DOI: 10.1371/journal.pone.0052089
    A phylogeographic study of an economically important freshwater fish, the striped snakehead, Channa striata in Sundaland was carried out using data from mtDNA ND5 gene target to elucidate genetic patterning. Templates obtained from a total of 280 individuals representing 24 sampling sites revealed 27 putative haplotypes. Three distinct genetic lineages were apparent; 1)northwest Peninsular Malaysia, 2)southern Peninsular, east Peninsular, Sumatra and SW (western Sarawak) and 3) central west Peninsular and Malaysian Borneo (except SW). Genetic structuring between lineages showed a significant signature of natural geographical barriers that have been acting as effective dividers between these populations. However, genetic propinquity between the SW and southern Peninsular and east Peninsular Malaysia populations was taken as evidence of ancient river connectivity between these regions during the Pleistocene epoch. Alternatively, close genetic relationship between central west Peninsular Malaysia and Malaysian Borneo populations implied anthropogenic activities. Further, haplotype sharing between the east Peninsular Malaysia and Sumatra populations revealed extraordinary migration ability of C. striata (>500 km) through ancient connectivity. These results provide interesting insights into the historical and contemporary landscape arrangement in shaping genetic patterns of freshwater species in Sundaland.
  10. Genotyping of microsatellite markers to study genetic structure of the wild striped snakehead Channa striata in Malaysia
    Tan MP, Jamsari AF, Siti Azizah MN
    J Fish Biol, 2016 May;88(5):1932-48.
    PMID: 27027270 DOI: 10.1111/jfb.12956
    Genetic variability and differences in wild striped snakehead Channa striata from Malaysia were analysed by genotyping nine novel nuclear microsatellite loci. Analysis revealed moderate-to-high genetic diversity in most of the populations, indicative of large effective population sizes. The highly diversified populations are admixed populations and, therefore, can be recommended as potential candidates for selective breeding and conservation since they each contain most of the alleles found in their particular region. Three homogenous groups of the wild populations were identified, apparently separated by effective barriers, in accordance with contemporary drainage patterns. The highest population pairwise FST found between members of the same group reflects the ancient population connectivity; yet prolonged geographical isolation resulted in adaptation of alleles to local contemporary environmental change. A significant relationship between genetic distance and geographical isolation was observed (r = 0·644, P < 0·01). Anthropogenic perturbations indicated apparent genetic proximity between distant populations.
  11. Morphological variation between isolates of the nematode Haemonchus contortus from sheep and goat populations in Malaysia and Yemen
    Gharamah AA, Rahman WA, Siti Azizah MN
    J Helminthol, 2014 Mar;88(1):82-8.
    PMID: 23176779 DOI: 10.1017/S0022149X12000776
    Haemonchus contortus is a highly pathogenic nematode parasite of sheep and goats. This work was conducted to investigate the population and host variations of the parasitic nematode H. contortus of sheep and goats from Malaysia and Yemen. Eight morphological characters were investigated, namely the total body length, cervical papillae, right spicule, left spicule, right barb, left barb, gubernaculum and cuticular ridge (synlophe) pattern. Statistical analysis showed the presence of morphological variation between populations of H. contortus from Malaysia and Yemen, with minor variation in the synlophe pattern of these isolates. Isolates from each country were grouped together in the scatterplots with no host isolation. Body, cervical papillae and spicule lengths were the most important characters that distinguished between populations of the two countries. This variation between Malaysia and Yemen may be attributed to geographical isolation and the possible presence of a different isolate of this worm in each country.
  12. Fulltext Genetic structure of the snakehead murrel, Channa striata (channidae) based on the cytochrome c oxidase subunit I gene: Influence of historical and geomorphological factors
    Jamsari AF, Jamaluddin JA, Pau TM, Siti-Azizah MN
    Genet Mol Biol, 2011 01;34(1):152-60.
    PMID: 21637559 DOI: 10.1590/S1415-47572011000100026
    Nucleotide sequences of a partial cytochrome c oxidase subunit I gene were used to assess the manner in which historical processes and geomorphological effects may have influenced genetic structuring and phylogeographic patterns in Channa striata. Assaying was based on individuals from twelve populations in four river systems, which were separated into two regions, the eastern and western, of the biodiversely rich state of Perak in central Peninsular Malaysia. In 238 specimens, a total of 368-bp sequences with ten polymorphic sites and eleven unique haplotypes were detected. Data on all the twelve populations revealed incomplete divergence due to past historical coalescence and the short period of separation. Nevertheless, SAMOVA and F(ST) revealed geographical structuring existed to a certain extent in both regions. For the eastern region, the data also showed that the upstream populations were genetically significantly different compared to the mid- and downstream ones. It is inferred that physical barriers and historical processes played a dominant role in structuring the genetic dispersal of the species. A further inference is that the Grik, Tanjung Rambutan and Sungkai are potential candidates for conservation and aquaculture programmes since they contained most of the total diversity in this area.
  13. Fulltext Evidence of geographical structuring in the Malaysian Snakehead, Channa striata based on partial segment of the CO1 gene
    Siti-Balkhis AB, Jamsari AF, Hwai TS, Yasin Z, Siti-Azizah MN
    Genet Mol Biol, 2011 Jul;34(3):520-3.
    PMID: 21931528 DOI: 10.1590/S1415-47572011005000016
    Channa striata, locally known as "haruan", is economically important in fisheries and aquaculture industries in several Asian countries. DNA sequencing, based on a partial segment of the Cytochrome oxidase c subunit 1 (CO1) gene, was used to determine genetic variation in C. striata samples from four different populations on the west coast of Peninsular Malaysia. The highest nucleotide and haplotype diversities were observed in the Linggi population (π = 0.0067, h = 0.835), and the lowest in the Timah Tasoh population (π = 0.0008, h = 0.286). Apart from Kajang-Linggi, which was insignificant, F(ST) values were significant (p < 0.05) in all pairwise-population comparisons. Consequently, it is inferred that genetic structuring C. striata populations in this region was largely shaped by a common origin, with secondary influences from geographical factors and isolation.
  14. Evaluation of mentum deformities of Chironomus spp. (Chironomidae: Diptera) larvae using modified toxic score index (MTSI) to assess the environmental stress in Juru River Basin, Penang, Malaysia
    Al-Shami SA, Salmah MR, Hassan AA, Azizah MN
    Environ Monit Assess, 2011 Jun;177(1-4):233-44.
    PMID: 20697808 DOI: 10.1007/s10661-010-1630-1
    Morphological mentum deformities which represent sublethal effect of exposure to different types of pollutants were evaluated in Chironomus spp. larvae inhabiting three polluted rivers of Juru River Basin in northwestern peninsular Malaysia. Using mentum deformity incidences, the modified toxic score index (MTSI) was developed based on Lenat's toxic score index (TSI). The suggested MTSI was compared with TSI in terms of its effectiveness to identify different pollutants including heavy metals. The MTSI showed stronger relationship to total deformity incidence expressed as percentage. Additionally, the multivariate RDA model showed higher capability of MTSI to explain the variations in heavy metal contents of the river sediments. The MTSI was recommended in bioassessment of water and sediment quality using the mentum deformities of Chironomus spp. larvae from aquatic ecosystems receiving anthropogenic, agricultural, or industrial discharges.
  15. Fulltext Hidden genetic diversity in snakeskin gourami, Trichopodus pectoralis (Perciformes, Osphronemidae), inferred from the mitochondrial DNA CO1 gene
    Tan MP, Amornsakun T, Siti Azizah MN, Habib A, Sung YY, Danish-Daniel M
    Mitochondrial DNA B Resour, 2019 Sep 12;4(2):2966-2969.
    PMID: 33365813 DOI: 10.1080/23802359.2019.1662741
    Eighty-four specimens collected from 13 populations from Malaysia, Thailand, and Vietnam were analysed, revealing 21 putative haplotypes with overall estimated haplotype and nucleotide diversities of 0.79 and 0.0079, respectively. High levels of diversity and an absence of founder effects were observed among populations in peninsular Malaysia. In contrast, populations from Sarawak exhibited low genetic diversity, which is a typical sign of colonies introduced from a single source. Historical translocation of Trichopodus pectoralis from Thailand to Malaysia, as well as to the Philippines, Indonesia, and Myanmar was apparent. Historical introduction of T. pectoralis from Vietnam was also detected in peninsular Malaysia.
  16. Fulltext Assessing the species composition of tropical eels (Anguillidae) in Aceh Waters, Indonesia, with DNA barcoding gene cox1
    Muchlisin ZA, Batubara AS, Fadli N, Muhammadar AA, Utami AI, Farhana N, et al.
    F1000Res, 2017;6:258.
    PMID: 28529702 DOI: 10.12688/f1000research.10715.1
    The objective of the present study was to evaluate the species diversity of eels native to Aceh waters based on genetic data. Sampling was conducted in western coast waters of Aceh Province, Indonesia, from July to August 2016. Genomic DNA was extracted from the samples, a genomic region from the 5' region of the cox1 gene was amplified and sequenced, and this was then used to analyse genetic variation. The genetic sequences were blasted into the NCBI database. Based on this analysis there were three valid species of eels that occurred in Aceh waters, namely Anguilla marmorata, A. bicolor bicolor, and A. bengalensis bengalensis.
  17. Effect of type and concentration of cryoprotectant on the motility, viability, and fertility of climbing perch Anabas testudineus Bloch, 1792 (Pisces: Anabantidae) sperm
    Maulida S, Eriani K, Fadli N, Kocabaş FK, Siti-Azizah MN, Wilkes M, et al.
    Theriogenology, 2023 Apr 15;201:24-29.
    PMID: 36822040 DOI: 10.1016/j.theriogenology.2023.02.014
    The climbing perch, Anabas testudineus is a freshwater fish that has economic value in Indonesia. It is cultured in the country, but the breeding technology, specifically sperm storage, is not well developed. Sperm cryopreservation is one of the preservation methods that need to be developed to support fish breeding technology. The type of cryoprotectants and its concentration are species-dependent and determines the success of this approach. Therefore, this study is aimed at determining the optimal type and concentration of cryoprotectant for sperm cryopreservation of A. testudineus. Four separate study series were performed, each of which evaluated one type of cryoprotectant at five concentration levels. The cryoprotectants used were DMSO, methanol, glycerol, and ethanol, and the tested concentrations were 0%, 5%, 10%, 15%, and 20%, which were combined with 5% egg yolks. Each treatment was conducted with three replications. The results showed that the type of cryoprotectant and its concentration significantly affected sperm motility, viability, and fertility of climbing perch (P 
  18. Fulltext Lernaea cyprinacea Linnaeus, 1758 (Copepoda: Lernaeidae) infection on Betta rubra Perugia, 1893 (Anabantiformes: Osphronemidae) from Aceh Province, Indonesia
    Nur FM, Batubara AS, Fadli N, Rizal S, Siti-Azizah MN, Wilkes M, et al.
    Rev Bras Parasitol Vet, 2022;31(1):e020421.
    PMID: 35293430 DOI: 10.1590/S1984-29612022015
    Betta rubra is an ornamental freshwater fish endemic to northern Sumatra, Indonesia. The B. rubra population has decreased in recent decades, and is classified as an endangered species in the IUCN Red List. This study aims to report for the first time infection by L. cyprinacea in B. rubra harvested from the Aceh Besar region of Indonesia. The fish samples were obtained from the Cot Bira tributaries, Aceh Besar District, Indonesia from January to December 2020. The results showed that the parasite infected 6 out of 499 samples in August and September, with a prevalence and intensity rate of 1% and 2 parasites/fish, respectively. The eyes and pectoral fins were the common infection sites. Despite B. rubra is not an optimal host (small size) for the parasite, this parasite might serve as additional threatening factors for the endangered B. rubra fish population.
  19. Fulltext Data of feed formulation for Indonesian short-fin eel, Anguilla bicolor McClelland, 1844 elver
    Muchlisin ZA, Sofyan M, Dewiyanti I, Nur FM, Batubara AS, Fadli N, et al.
    Data Brief, 2020 Jun;30:105581.
    PMID: 32346582 DOI: 10.1016/j.dib.2020.105581
    This article describes the data of raw material compositions for formulating the diet for Indonesian short-fin eel, Anguilla bicolor Anguilla bicolor McClelland, 1844 elver. The data on growth performance, survival and feed utilization of eel elver from the formulated diet test results are also presented here. Four formulated diets and one commercial feed were tested for the eel elver. The formulated diet was formulated from animal and plant based protein sources such as; fish meal, cornmeal, soybean meal, fine bran, ebi-shrimp meal, golden snail meal, bloodmeal, and tapioca flour. Vitamins and minerals were added into the diet. In addition, probiotics and papain enzyme and its combination were also included into the experimental diets to boost the growth performance, survival and feed utilization of the fish. The tested treatments were; (A) commercial diet without probiotic and papain enzyme, (B) experimental diet without probiotic and papain enzyme, (C) experimental diet with probiotic 10 ml kg-1 feed, (D) experimental diet with papain enzyme 10 g kg-1 feed, (E) experimental diet with probiotic 10 ml kg-1 + papain 10 g kg-1 feed, (F) experimental diet with probiotic 5 ml kg-1 + papain 5 g kg-1 feed. Each experimental fish group was fed on an experimental diet twice a day at 08.00 AM and 06.00 PM at feeding level of 10% body weight for 60 days. The results showed that the weight gain ranged between 4.60 g to 5.92 g, daily growth rate between 0.063 g day-1 and 0.098 g per day-1, the specific growth rate varies from 0.72% day-1 to 1.35% day-1, survival rate 50.00% to 73.33%, feed conversion ratio from 2.35 to 4.17, and feed efficiency between 41.82% to 58.98%. The best treatment was found in fish fed on the experimental diet with papain enzyme 10 g kg-1 of feed.
  20. Motility, viability and fertility of goldfish Carassius auratus (Pisces: Cyprinidae) post short-term cryopreservation
    Nurlaili N, Eriani K, Salma I, Maulida S, Rahayu SR, Handayani LS, et al.
    Cryo Letters, 2023;44(3):169-177.
    PMID: 37883170
    BACKGRUND: Goldfish Carassius auratus is a popular ornamental fish extensively cultured worldwide. Sperm cryopreservation is a common fish breeding method that ensures sperm availability around the year. Studies on cryopreservation of goldfish sperm, especially on the suitability of cryoprotectant types and pre-freezing time, are scarcely available.

    OBJECTIVE: To determine the most suitable type of cryoprotectant and pre-freezing for the successful cryopreservation of goldfish sperm.

    MATERIALS AND METHODS: A completely randomized design with two factors was utilized in this study. The first factor is the type of cryoprotectants, which included methanol, ethanol, ethylene glycol, glycerol, and DMSO. The second is pre-freezing times of 10, 20, 30, and 40 min at each of the pre-freezing temperatures of 4 degree C, -10 degree C, and -79 degree C, meaning that the total times for the ramping down of temperature were 30, 60, 90 and 120 min, respectively. The Ringer solution and 10% egg yolk were used as extender and extracellular cryoprotectant. The sperm was stored at -179 degree C for 7 days.

    RESULTS: The ANOVA test showed that cryoprotectants and pre-freezing significantly affected the motility, viability, and fertility of goldfish sperm after freezing in liquid nitrogen for 7 days (P<0.05). Furthermore, 10% DMSO combined with 15% egg yolk with an pre-freezing time of 20 min can maintain sperm motility, viability, and fertility higher than other treatments, by 79%, 80%, and 33%, respectively. The agarose gel electrophoresis showed no DNA fragmentation in all samples, including fresh sperm.

    CONCLUSION: We conclude that 10% DMSO combined with 15% egg yolk and 20 min pre-freezing is the best treatment for goldfish sperm cryopreservation. DOI: 10.54680/fr23310110412.

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