scholarly journals Prevalence of bovine astroviruses and their genotypes in sampled Chinese calves with and without diarrhoea

2021 ◽  
Vol 102 (8) ◽  
Author(s):  
Jie Zhu ◽  
Mingpu Qi ◽  
Chuanwen Jiang ◽  
Yongchong Peng ◽  
Qingjie Peng ◽  
...  
Keyword(s):  
Genetic Diversity ◽  
Binary Logistic Regression ◽  
Binary Logistic Regression Analysis ◽  
Strong Positive Correlation ◽  
High Genetic Diversity ◽  
Bovine Rotavirus ◽  
Calf Diarrhoea ◽  
Faecal Samples ◽  
Polymerase Chain ◽  
Next Generation Sequencing Ngs

Bovine astrovirus (BoAstV) belongs to genus Mamastravirus (MAstV). It can be detected in the faeces of both diarrhoeal and healthy calves. However, its prevalence, genetic diversity, and association with cattle diarrhoea are poorly understood. In this study, faecal samples of 87 diarrhoeal and 77 asymptomatic calves from 20 farms in 12 provinces were collected, and BoAstV was detected with reverse transcription-polymerase chain reaction (RT-PCR). The overall prevalence rate of this virus in diarrhoeal and asymptomatic calves was 55.17 % (95 % CI: 44.13, 65.85 %) and 36.36 % (95 % CI: 25.70, 48.12 %), respectively, indicating a correlation between BoAstV infection and calf diarrhoea (OR=2.15, P=0.024). BoAstV existed mainly in the form of co-infection (85.53 %) with one to five of nine viruses, and there was a strong positive correlation between BoAstV co-infection and calf diarrhoea (OR=2.83, P=0.004). Binary logistic regression analysis confirmed this correlation between BoAstV co-infection and calf diarrhoea (OR=2.41, P=0.038). The co-infection of BoAstV and bovine rotavirus (BRV) with or without other viruses accounted for 70.77 % of all the co-infection cases. The diarrhoea risk for the calves co-infected with BoAstV and BRV was 8.14-fold higher than that for the calves co-infected with BoAstV and other viruses (OR=8.14, P=0.001). Further, the co-infection of BoAstV/BRV/bovine kobuvirus (BKoV) might increase the risk of calf diarrhoea by 14.82-fold, compared with that of BoAstV and other viruses (OR=14.82, P <0.001). Then, nearly complete genomic sequences of nine BoAstV strains were assembled by using next-generation sequencing (NGS) method. Sequence alignment against known astrovirus (AstV) strains at the levels of both amino acids and nucleotides showed a high genetic diversity. Four genotypes were identified, including two known genotypes MAstV-28 (n=3) and MAstV-33 (n=2) and two novel genotypes designated tentatively as MAstV-34 (n=1) and MAstV-35 (n=3). In addition, seven out of nine BoAstV strains showed possible inter-genotype recombination and cross-species recombination. Therefore, our results increase the knowledge about the prevalence and the genetic evolution of BoAstV and provide evidence for the association between BoAstV infection and calf diarrhoea.

scholarly journals Microsatellite genotyping from faeces of Lontra longicaudis from southern Brazil

2009 ◽  
Vol 99 (1) ◽  
pp. 5-11 ◽  
Author(s):  
Laura I. Weber ◽  
Cintia G. Hildebrand ◽  
Anderson Ferreira ◽  
Gustavo Pedarassi ◽  
José A. Levy ◽  
...  
Keyword(s):  
Genetic Diversity ◽  
Conservation Status ◽  
Significant Degree ◽  
Southern Brazil ◽  
High Genetic Diversity ◽  
Low Degree ◽  
Faecal Samples ◽  
High Level ◽  
Degree Of Differentiation ◽  
Lontra Longicaudis

A genetic study of the neotropical river otter Lontra longicaudis (Olfers, 1818), which has an unknown conservation status, was carried out at the Taim Ecological Station and the margins of the Vargas stream, Rio Grande do Sul, southern Brazil. Faecal samples were collected, and DNA was extracted using a silica-guanidine method. Five microsatellite loci were amplified using PCR with heterologous primers previously described for Lutra lutra (Linnaeus, 1758). Sixteen faecal samples out of 29 from Taim and 11 out of 14 from Vargas stream margins contained enough DNA for genetic analysis. A total of 49 different alleles were found at both localities, from which 18 were exclusively found in individuals from Taim and 17 were exclusives from Vargas individuals. The most common allele was the same at both locations for three loci (Lut715, Lut733, and Lut818). A high level of genetic diversity was found at both sites (NeTaim=4.1, HoTaim=0.299, HeTaim=0.681; NeVargas=4.9, HoVargas=0.355, HeVargas=0.724), being higher at the Vargas stream site. A high and significant level of heterozygote deficiency was observed at most loci according to the χ2 test. The homogeneity χ2 test (P<0.001) showed that there were significant differences in the allele frequencies between the two locations. Genotyping for more than one locus was possible in 81.5% of samples, from which only 37% were possible to genotype for more than three loci. A low degree of relatedness was found among individuals from Taim (R=0.055±0.310), but an even lower value of relatedness was found at the Vargas site (R= -0.285±0.440). The significant degree of differentiation (I=0.890; F ST=0.059) found between Taim and Vargas individuals suggests that there is more than one population of otters in the southern extreme of Brazil, which probably are associated with the water body systems found in this region, the Mirim and the Caiuvá/Flores/Mangueira Lagoons. The high genetic diversity and low relatedness found at the Vargas stream, lead us to believe that the Vargas stream may be acting as a corridor between these water bodies for otter dispersion.

scholarly journals Astrovirus in Reunion Free-Tailed Bat (Mormopterus francoismoutoui)

Viruses ◽  
2021 ◽  
Vol 13 (8) ◽  
pp. 1524
Author(s):  
Léa Joffrin ◽  
Axel O. G. Hoarau ◽  
Erwan Lagadec ◽  
Marie Köster ◽  
Riana V. Ramanantsalama ◽  
...  
Keyword(s):  
Genetic Diversity ◽  
Rna Viruses ◽  
Mammalian Species ◽  
Rdrp Gene ◽  
Chain Reaction ◽  
Rna Dependent Rna Polymerase ◽  
Nested Polymerase Chain Reaction ◽  
High Genetic Diversity ◽  
Partial Rdrp ◽  
Polymerase Chain

Astroviruses (AstVs) are RNA viruses infecting a large diversity of avian and mammalian species, including bats, livestock, and humans. We investigated AstV infection in a free-tailed bat species, Mormopterus francoismoutoui, endemic to Reunion Island. A total of 380 guano samples were collected in a maternity colony during 38 different sampling sessions, from 21 June 2016 to 4 September 2018. Each sample was tested for the presence of the AstV RNA-dependent RNA-polymerase (RdRp) gene using a pan-AstV semi-nested polymerase chain reaction assay. In total, 27 guano samples (7.1%) tested positive, with high genetic diversity of the partial RdRp gene sequences among positive samples. Phylogenetic analysis further revealed that the detected viruses were genetically related to AstVs reported in rats, reptiles, dogs, and pigs, but did not cluster with AstVs commonly found in bats. Although more investigations need to be conducted to assess the prevalence of infected bats in the studied population, our findings show that Reunion free-tailed bats are exposed to AstVs, and suggest that cross-species transmission may occur with other hosts sharing the same habitat.

scholarly journals Diversity Analysis and Genetic Relationships among Local Brazilian Goat Breeds Using SSR Markers

Animals ◽  
2020 ◽  
Vol 10 (10) ◽  
pp. 1842
Author(s):  
Marcos Paulo Carrera Menezes ◽  
Amparo Martinez Martinez ◽  
Edgard Cavalcanti Pimenta Filho ◽  
Jose Luis Vega-Pla ◽  
Juan Vicente Delgado ◽  
...  
Keyword(s):  
Genetic Diversity ◽  
Genetic Relationships ◽  
Fixation Index ◽  
High Genetic Diversity ◽  
Allele Number ◽  
Hair Samples ◽  
The Mean ◽  
Polymerase Chain ◽  
Single Sequence ◽  
Brazilian Populations

The genetic diversity of six Brazilian native goats was reported using molecular markers. Hair samples of 332 animals were collected from different goat breeds (Moxotó, Canindé, Serrana Azul, Marota, Repartida, and Graúna) from five states of Northeast Brazil (Paraíba, Pernambuco, Rio Grande do Norte, Bahia, and Piauí). A panel of 27 microsatellites or single sequence repeats (SSRs) were selected and amplified using a polymerase chain reaction (PCR) technique. All populations showed an average allele number of over six. The mean observed heterozygosity for Brazilian breeds was superior to 0.50. These results demonstrated the high genetic diversity in the studied populations with values ranging from 0.53 (Serrana Azul) to 0.62 (Repartida). The expected average heterozygosity followed the same trend ranging from 0.58 (Serrana Azul) to 0.65 (Repartida), and the values obtained are very similar for all six breeds. The fixation index (Fis) had values under 10% except for the Moxotó breed (13%). The mean expected heterozygosity of all Brazilian populations was over 0.50. Results indicated a within-breed genetic variability in the Brazilian breeds based on the average number of alleles and the average observed heterozygosity. The interbreed genetic diversity values showed proper genetic differentiation among local Brazilian goat breeds.

scholarly journals ITGA4, ITGB7, TNFα, IL10 genes polymorphisms in the ethnic Buryat patients with ulcerative colitis

2021 ◽  
Vol 49 ◽  
Author(s):  
I. V. Zhilin ◽  
E. Yu. Chashkova ◽  
A. A. Zhilina ◽  
A. Ch. Tsyrempilova
Keyword(s):  
Ulcerative Colitis ◽  
Healthy Volunteers ◽  
Molecular Genetic ◽  
Genetic Material ◽  
Binary Logistic Regression ◽  
Molecular Genetic Analysis ◽  
Binary Logistic Regression Analysis ◽  
Irkutsk Region ◽  
In Trans ◽  
Polymerase Chain

Background: Worldwide studies of genetic material, polymorphisms and prognostic gene models for immune-associated disorders have established differences in trans-ethnic population cohorts, which determine phenotypic and other characteristics of the course of these diseases. Ulcerative colitis (UC) is a  chronic immune inflammation of the colon mucosa. More than 100 gene polymorphisms associated with multiple integrated cross-talks have been discovered.Aim: To study the ITGA4, ITGB7, TNFα, IL10 genes polymorphisms in patients with ulcerative colitis belonging to the Buryat ethnic group and living in Irkutsk region, Buryat Republic and Transbaikal territory.Materials and methods: The study included a total of 49 subjects, 24 of them being UC patients and 25 healthy volunteers, compatible in gender, age and ethnic background. The molecular genetic analysis by real time polymerase chain reaction was performed with DNA samples from whole peripheral blood leucocytes.Results: The differences in the prevalence of the ITGA4(rs1143674, rs1449263), ITGB7(rs11574532), TNFα(rs1800629), and IL10(rs1800871) genotypes were non-significant (р>0.05). The IL10(rs1800896) GG homozygote patients had higher odds ratio (OR) for UC compared to the carriers of other polymorphisms (OR 24; 95%  confidence interval (CI) 2.783–206.969; р=0.001). The AA homozygote type was less frequent among UC patients compared to healthy volunteers (OR 0.17; 95%  CI 0.049–0.589; р=0.004). The analysis of genotype frequency distribution of all studied genes including clinical characteristics of the disease showed no significant results (р>0.05). The binary logistic regression analysis has shown that IL10(rs1800896)GG was an UC predictor with sensitivity of 96% and specificity of 50%  (AUC 0.760; 95% CI 0.621–0.899; p=0.002; standard error 0.71).Conclusion: The GG genotype of IL10(rs1800896) is a  UC predictor, whereas the AA genotype is significantly more prevalent among healthy subjects of the Buryat cohort. 

scholarly journals Astrovirus in Reunion Free-tailed Bat (Mormopterus francoismoutoui)

2019 ◽  
Author(s):  
Léa Joffrin ◽  
Axel O. G. Hoarau ◽  
Erwan Lagadec ◽  
Patrick Mavingui ◽  
Camille Lebarbenchon
Keyword(s):  
Genetic Diversity ◽  
Rna Viruses ◽  
Mammalian Species ◽  
Rdrp Gene ◽  
Chain Reaction ◽  
Rna Dependent Rna Polymerase ◽  
Nested Polymerase Chain Reaction ◽  
High Genetic Diversity ◽  
Partial Rdrp ◽  
Polymerase Chain

AbstractAstroviruses (AstVs) are RNA viruses responsible for infection of a large diversity of avian and mammalian species, including bats, livestock, and humans. We investigated AstV infection in a free-tailed bat species, Mormopterus francoismoutoui, endemic to Reunion Island. A total of 190 guano samples were collected in a maternity colony during 19 different sampling sessions, between June 2016 and June 2017. Biological material was tested for the presence of the AstV RNA-dependent RNA-polymerase (RdRp) gene with a pan-AstV semi-nested polymerase chain reaction assay. In total, 15 guano samples (7.9%) tested positive, with high genetic diversity of the partial RdRp gene sequences among positive samples. A phylogenetic analysis further revealed that the detected viruses were genetically related to AstVs reported in reptiles, dogs, and pigs, but did not cluster with AstVs commonly found in bats. Although more investigation need to be conducted to assess the level of infected bats in the studied population, our findings suggest that Reunion free-tailed bats are exposed to AstV, and that cross-species transmission may occur with other hosts sharing the same habitat.

Assessment of molecular diversity in landraces of bread wheat (Triticum aestivum L.) held in an ex situ collection with Diversity Arrays Technology (DArT™)

2007 ◽  
Vol 58 (12) ◽  
pp. 1174 ◽  
Author(s):  
B. J. Stodart ◽  
M. C. Mackay ◽  
H. Raman
Keyword(s):  
Genetic Diversity ◽  
Molecular Diversity ◽  
Polymorphic Information Content ◽  
Ex Situ ◽  
Dart Markers ◽  
Strong Positive Correlation ◽  
High Genetic Diversity ◽  
Diversity Arrays Technology ◽  
Wheat Landraces ◽  
Landrace Accessions

Diversity Arrays Technology (DArT™) was evaluated as a tool for determining molecular diversity of wheat landraces held within the Australian Winter Cereals Collection (AWCC). Initially, a set of 44 wheat landraces was evaluated with 256 DArT markers. The dataset was compared with the results obtained using 16 amplified fragment length polymorphism (AFLP) primer combinations and 63 simple sequence repeat (SSR) markers, mapped on the 21 chromosomes, from a previous study. The DArT markers exhibited a strong positive correlation with AFLP and SSR, with each marker type distinguishing similar relationships among the 44 landrace accessions. The DArT markers exhibited a higher polymorphic information content than AFLP, and were comparable with that obtained with SSR. Three hundred and fifty-five DArT markers were then used to evaluate genetic diversity among 705 wheat landrace accessions from within the AWCC, chosen to represent 5 world regions. DArT analysis was capable of distinguishing accessions from different geographic regions, and suggested that accessions originating from Nepal represent a unique gene pool within the collection. A statistical resampling of DArT loci indicated that 10–20 loci were enough to distinguish the maximum molecular diversity present within the collection. This research demonstrates the efficacy of the DArT platform as a tool for efficient examination of wheat diversity. As an ex situ germplasm repository, the AWCC contains wheat accessions of high genetic diversity, from genetically differentiated collection sites, even though diversity was under-represented in some countries represented in the repository.

scholarly journals Molecular detection and characterisation of sapoviruses and noroviruses in outpatient children with diarrhoea in Northwest Ethiopia

2019 ◽  
Vol 147 ◽  
Author(s):  
A. Gelaw ◽  
C. Pietsch ◽  
P. Mann ◽  
U.G. Liebert
Keyword(s):  
Genetic Diversity ◽  
Low Income ◽  
Northwest Ethiopia ◽  
Low Income Countries ◽  
African Countries ◽  
High Genetic Diversity ◽  
Childhood Morbidity ◽  
Faecal Samples ◽  
Stool Samples ◽  
Outpatient Children

Abstract Childhood morbidity and mortality of diarrhoeal diseases are high, particularly in low-income countries and noroviruses and sapoviruses are among the most frequent causes worldwide. Their epidemiology and diversity remain not well studied in many African countries. To assess the positivity rate and the diversity of sapoviruses and noroviruses in Northwest Ethiopia, during November 2015 and April 2016, a total of 450 faecal samples were collected from outpatient children aged <5 years who presented with diarrhoea. Samples were screened for noroviruses and sapoviruses by real-time RT-PCR. Partial VP1 genes were sequenced, genotyped and phylogenetically analysed. Norovirus and sapovirus stool positivity rate was 13.3% and 10.0%, respectively. Noroviruses included GII.4 (35%), GII.6 (20%), GII.17 (13.3%), GII.10 (10%), GII.2 (6.7%), GII.16 (5%), GII.7 (3.3%), GII.9, GII.13, GII.20 and GI.3 (1.7% each) strains. For sapoviruses, GI.1, GII.1 (20.0% each), GII.6 (13.3%), GI.2 (8.9%), GII.2 (11.1%), GV.1 (8.9%), GIV.1 (6.7%), GI.3 and GII.4 (2.2% each) genotypes were detected. This study demonstrates a high genetic diversity of noroviruses and sapoviruses in Northwest Ethiopia. The positivity rate in stool samples from young children with diarrhoea was high for both caliciviruses. Continued monitoring is recommended to identify trends in genetic diversity and seasonal variations.

scholarly journals Species distribution, antibiotic resistance and virulence traits in canine and feline enterococci in Tunisia

2017 ◽  
Vol 65 (2) ◽  
pp. 173-184 ◽  
Author(s):  
Leila Ben Said ◽  
Raoudha Dziri ◽  
Nadia Sassi ◽  
Carmen Lozano ◽  
Karim Ben Slama ◽  
...  
Keyword(s):  
Genetic Diversity ◽  
Gel Electrophoresis ◽  
Virulence Genes ◽  
Pulsed Field Gel Electrophoresis ◽  
High Genetic Diversity ◽  
Antibiotic Resistant ◽  
Faecal Samples ◽  
Household Environment ◽  
High Level

In order to investigate the possible role of dogs and cats in the carriage and potential dissemination of resistant enterococci, seventy faecal samples from dogs and cats were tested for enterococci. Fifty-eight enterococci were recovered. Isolates were identified as Enterococcus faecium (n = 31) and E. faecalis (n = 14) E. durans (n = 6), E. casseliflavus (n = 2), E. hirae and E. gallinarum (2 isolates each). Enterococcal isolates showed resistance to ciprofloxacin (n = 35), erythromycin (n = 31), tetracycline (n = 25), kanamycin (n = 15), streptomycin (n = 13), pristinamycin (n = 11), gentamicin (n = 10), chloramphenicol (n = 8), and linezolid (n = 6). The gene erm(B) was detected in 22 out of 31 erythromycin-resistant enterococci. All tetracycline-resistant enterococci carried tet(M) and/or tet(L) genes. The gene aac(6′)-Ie-aph(2″)-Ia was identified in five of high-level gentamicin-resistant isolates, the genes aph(3′)-IIIa and/or aac(6′)-Ie-aph(2″)-Ia in eleven high-level kanamycin-resistant isolates and the gene ant(6)-Ia in eleven high-level streptomycin-resistant isolates. Only one strain harboured cat(A) gene, and five strains contained vat(E) or vat(D) genes. Virulence genes gel(E) (21 strains), esp (11 strains) and cylA/cylB (5 strains) were detected. High genetic diversity was demonstrated among E. faecium isolates by pulsed-field gel electrophoresis (PFGE). Dogs and cats can be carriers of antibiotic-resistant enterococci in their faeces that could shed into the household environment.

scholarly journals Genotypic analyses and virulence characterization ofGlaesserella parasuisisolates from Taiwan

PeerJ ◽  
2019 ◽  
Vol 7 ◽  
pp. e6960 ◽  
Author(s):  
Wei-Hao Lin ◽  
Hsing-Chun Shih ◽  
Chuen-Fu Lin ◽  
Cheng-Yao Yang ◽  
Chao-Nan Lin ◽  
...  
Keyword(s):  
Genetic Diversity ◽  
Partial Correlation ◽  
Economic Losses ◽  
Swine Industry ◽  
High Genetic Diversity ◽  
Molecular Serotyping ◽  
Positive Rate ◽  
Polymerase Chain ◽  
Different Strains ◽  
Group 1

BackgroundGlaesserella(Haemophilus)parasuis(G. parasuis) causes severe economic losses in the swine industry. MultipleG. parasuisstrains can exist in single animals. Typing techniques are required for identifyingG. parasuisisolates. Different strains within a serovar display varying virulence. Enterobacterial repetitive intergenic consensus polymerase chain reaction (ERIC-PCR) can assess the heterogeneity. The group 1 virulence-associated trimeric autotransporters (vtaA) gene is an indicator of virulence. The aim of this study was to characterize TaiwaneseG. parasuisisolates via molecular serotyping, vtaA PCR and ERIC-PCR.MethodsOne hundred and forty-five strains were collected between November 2013 and March 2017 in Taiwan and further examined by molecular serotyping, vtaA PCR and ERIC-PCR.ResultsThe dendrogram revealed heterogeneous genetic diversity within many clusters. Partial correlation between the ERIC-PCR clusters of different strains, serovars and lesion patterns was observed. Twelve herds (8.3%) infected with more than one strain. Group 1 vtaA positive rate reached 98.6%.DiscussionThis study showed the high genetic diversity ofG. parasuisin Taiwan by a high discriminatory capability of ERIC-PCR. Group 1 vtaA commonly exists inG. parasuisisolates and may play important roles in the pathogenesis of TaiwaneseG. parasuisisolates.

scholarly journals Keragaman genetik ikan tuna sirip kuning (Thunnus albacares) dari dua populasi di Laut Maluku, Indonesia

DEPIK ◽  
2014 ◽  
Vol 3 (1) ◽  
Author(s):  
Nebuchadnezzar Akbar ◽  
Neviaty P Zamani ◽  
Hawis H Madduppa
Keyword(s):  
Genetic Diversity ◽  
Nucleotide Diversity ◽  
Haplotype Diversity ◽  
Yellowfin Tuna ◽  
Conservation Strategy ◽  
High Genetic Diversity ◽  
Tissue Samples ◽  
Chain Reactions ◽  
Polymerase Chain ◽  
Two Populations

Abstract. Yellowfin tuna (Thunnus albacores) is a large, pelagic, and migratory species of tuna that inhabits Moluccas Sea in Indonesia, and most sea environment worldwide. However, high fishing activities tend to happen in the Indonesia region and catch product appear to be decreasing. A better understanding of yellowfin tuna genetic diversity is required to plan better conservation strategy of tuna. The study was conducted to infer the genetic diversity of yellowfin tuna (T. albacores) in the Moluccas Sea. A total of 41 tissue samples of yellowfin tuna were collected from two regions in the Moluccas Sea (North Moluccas and Ambon) during an expedition in February 2013. The results showed that genetic diversity and nucleotide diversity of yellowfin tuna from North Moluccas population was 0.984 and 0.021, respectively; while in Ambon population, the genetic and nucleotide diversities were 1.00 and 0.018, respectively. The high genetic diversity (0.990) and nucleotide diversity (0.020) between two populations were observed. Based on phylogenetic analysis, no genetic differentiation between the two populations in Moluccas Sea was revealed .Keywords :  Population genetics; Haplotype diversity; Coral Triangle; Phylogenetics; Polymerase Chain Reactions (PCRs) Abstrak. Tuna sirip kuning (Thunnus albacores) adalah ikan komersial penting dan ditemukan di Laut Maluku, Indonesia. Tetapi, aktivitas penangkapan ikan tuna sirip kuning dapat menurunkan kualitas dan kuantitas stok ikan, sehingga perlu adanya pengkajian keragaman genetik ikan tuna sirip kuning. Pemahaman yang baik tentang keragaman genetika dibutuhkan untuk merencanakan strategi konservasi tuna yang lebih baik. Penelitian ini bertujuan untuk mengetahui keragaman genetik  ikan tuna sirip kuning dari dua populasi di Laut Maluku. Sebanyak 41 sampel jaringan dari tuna sirip kuning dikumpulkan dari dua populasi di Laut Maluku (Maluku Utara dan Ambon) selama ekspedisi pada bulan Februari 2013. Hasil penelitian menunjukkan bahwa keragaman genetik populasi tuna sirip kuning pada perairan Maluku Utara dan Ambon masing-masing sebesar 0,984 dan 1,00 sedangkan nilai keragaman nukleotida masing-masing bernilai 0,021 dan 0,018. Nilai keragaman genetik dan keragaman nukloetida yang tinggi didaptkan antar kedua populasi masing-masing sebesar 0,990 dan 0,020. Berdasarkan analisis filogenetik, dua populasi di Laut Maluku ini memiliki kedekatan secara genetik.Kata kunci : Genetika populasi; Keragaman haplotipe; Segitiga Terumbu Karang; Filogenetika; Polymerase Chain Reaction (PCR)

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