Moch Syamsul Arifin Zein

Selection of Mx gene genotype as genetic marker for Avian Influenza resistance in Indonesian native chicken

BackgroundIn previous studies, the Mx Gene has been demonstrated to confer positive anti viral responses in chicken. The amino acid variation of Asn (allele A) at position 631 was specific to positive antiviral Mx/resistant, while, that of Ser (allele G) was specific to negative Mx/susceptible. This research was aimed at selecting one of the native chicken breeds which was found out to be resistant to avian influenza using molecular technique. The selected breed will then be used as the base population to improve native chicken breed in Indonesia.MethodsMarker Assisted Selection (MAS) method was used in this research to accelerate the selection process, since the disease resistance had low heritability value. Polymerase Chain Reaction-Restriction Fragment Length Polymorphism (PCR-RFLP) technique used to select the genotype of Mx++, Mx+- and Mx-- that corresponded to the positive antiviral activity (Mx++), or those which had positive or negative activity (Mx+-) and negative antiviral activity (Mx--). There were 200 native hens and 40 cocks used in this experiment. Allele frequency of Mx Gene was calculated. The productivity indicators such as age at first laying, egg weight and hen weight at first laying and egg production were also measured. The chicken that had Mx++ and Mx+- genotypes, were selected to produce offspring.ResultsResult showed that the frequency of the resistant allele (Mx+) was 65% and 60% in laying hens and in cocks, respectively, while the frequency of the susceptible allele (Mx-) was 35% and 40% in hens and cocks, resepctively. Age, egg weight and hen weight at first laying and egg production for susceptible genotype were slightly better than for the resistant genotype which were 172,41 VS 178,81 days; 33,94 VS 32,84 g; 1450 VS 1439 g and 54,32 VS 48,30 %, respectively.

Development of a multiplex PCR assay for fine-scale population genetic analysis of the Komodo monitor Varanus komodoensis based on 18 polymorphic microsatellite loci

Multiplex PCR assays for the coamplification of microsatellite loci allow rapid and cost‐effective genetic analyses and the production of efficient screening protocols for international breeding programs. We constructed a partial genomic library enriched for di‐nucleotide repeats and characterized 14 new microsatellite loci for the Komodo monitor (or Komodo dragon, Varanus komodoensis). Using these novel microsatellites and four previously described loci, we developed multiplex PCR assays that may be loaded on a genetic analyser in three separate panels. We tested the novel set of microsatellites for polymorphism using 69 individuals from three island populations and evaluated the resolving power of the entire panel of 18 loci by conducting (i) a preliminary assignment test to determine population(s) of origin and (ii) a parentage analysis for 43 captive Komodo monitors. This panel of polymorphic loci proved useful for both purposes and thus can be exploited for fine‐scale population genetic analyses and as part of international captive breeding programs directed at maintaining genetically viable ex situ populations and reintroductions.

Study of Genetic Diversity among Simmental Cross Cattle in West Sumatra Based on Microsatellite Markers

A study was conducted to assess the genetic diversity among Simmental Cross cattle in West Sumatra using microsatellite DNA markers. A total of 176 individual cattle blood samples was used for obtaining DNA samples. Twelve primers of microsatellite loci as recommended by FAO were used to identify the genetic diversity of the Simmental Cross cattle population. Multiplex DNA fragment analysis method was used for allele identification. All the microsatellite loci in this study were highly polymorphic and all of the identified alleles were able to classify the cattle population into several groups based on their genetic distance. The heterozygosity values of microsatellite loci in this study ranged from 0.556 to 0.782. The polymorphism information content (PIC) value of the 12 observed loci is high (PIC>0.5). The highest PIC value in the Simmental cattle population was 0.893 (locus TGLA53), while the lowest value was 0.529 (locus BM1818). Based on the genetic distance value, the subpopulation of the Simmental Cross-Agam and the Simmental Cross-Limapuluh Kota was exceptionally close to the Simmental Purebred thus indicating that a grading-up process has taken place with the Simmental Purebred. In view of the advantages possessed by the Simmental Cross cattle and the evaluation of the genetic diversity results, a number of subpopulations in this study can be considered as the initial (base) population for the Simmental Cross cattle breeding programs in West Sumatra, Indonesia.

Genetic Diversity of Indonesian Cattle Breeds Based on Microsatellite Markers

Objective This research was conducted to study the genetic diversity in several Indonesian cattle breeds using microsatellite markers to classify the Indonesian cattle breeds. Methods A total of 229 DNA samples from of 10 cattle breeds were used in this study. The polymerase chain reaction process was conducted using 12 labeled primers. The size of allele was generated using the multiplex DNA fragment analysis. The POPGEN and CERVUS programs were used to obtain the observed number of alleles, effective number of alleles, observed heterozygosity value, expected heterozygosity value, allele frequency, genetic differentiation, the global heterozygote deficit among breeds, and the heterozygote deficit within the breed, gene flow, Hardy-Weinberg equilibrium, and polymorphism information content values. The MEGA program was used to generate a dendrogram that illustrates the relationship among cattle population. Bayesian clustering assignments were analyzed using STRUCTURE program. The GENETIX program was used to perform the correspondence factorial analysis (CFA). The GENALEX program was used to perform the principal coordinates analysis (PCoA) and analysis of molecular variance. The principal component analysis (PCA) was performed using adegenet package of R program. Results A total of 862 alleles were detected in this study. The INRA23 allele 205 is a specific allele candidate for the Sumba Ongole cattle, while the allele 219 is a specific allele candidate for Ongole Grade. This study revealed a very close genetic relationship between the Ongole Grade and Sumba Ongole cattle and between the Madura and Pasundan cattle. The results from the CFA, PCoA, and PCA analysis in this study provide scientific evidence regarding the genetic relationship between Banteng and Bali cattle. According to the genetic relationship, the Pesisir cattle were classified as Bos indicus cattle. Conclusion All identified alleles in this study were able to classify the cattle population into three clusters i.e. Bos taurus cluster (Simmental Purebred, Simmental Crossbred, and Holstein Friesian cattle); Bos indicus cluster (Sumba Ongole, Ongole Grade, Madura, Pasundan, and Pesisir cattle); and Bos javanicus cluster (Banteng and Bali cattle).

Aplikasi Kajian DNA Molekuler dan Fenotipik Pada Program Pelepasliaran Burung Kakatua

There are six species of cockatoos in Indonesia which are well known as exotic, smart, and they can be trained in a variety of attractions. Thus, many people want to keep those birds as pets. All of pets which have been kept by community should be evaluated from various aspects before being reintroduced to their natural habitat. The examination of sex and species of illegal cocktoos play as a key role for the reintroduction programme. Theobjective of this study was to evaluate the reliability and effectivity of evaluation technique of morphometric and molecular for reintroduction programme of cockatoos. We used the COI gene sequences from 68individuals of cockatoos from pet communities in and around Jakarta and four sequences from GenBank. The phylogenetic analysis used the neighbor-joining method, in which the genetic distance matrix calculations with Kimura 2-parameter models that are implemented on a pairwise distance calculation in the MEGA program version 6:05. The result of the genetic variation of the cockatoo species which shows intraspecific divergence was Cacatua alba (n=4)= 0%, C. galerita (Australia n=9)= 0.6%, C. galerita (Indonesia n=53)= 0.3%, C .goffiniana (n=3)= 0%, C. moluccensis (n=7)= 0.1%, and C. sulphurea (n=2)= 0.3%, with a range of 0-0.6%. The results indicate that the average of intraspecific of COI in the cockatoos community was 0.25±0.055%, and interspecific divergences ranged from 3.1 to 11.6%. The phylogenetic tree shows the monophyletic clade of cockatoo species in Indonesia. In addition, DNA barcode analysis and molecular sexing could correct the error and doubts the result of five individual species identification and two individual sexing identification of C.galerita by morphological identification. The results of morphological examanation base on body weight, body length and head-bill length of C. galerita triton were not significantly different (P?0,5). Finally, 19 individuals C. galerita triton and two individuals P. aterrimus were reintroduced to their natural habitat.