Background and Aim: Bovine eimeriosis is a disease caused by apicomplexan parasites of the genus Eimeria. It is one of the most important and widespread bovine illnesses in the world. Some of the identified species of bovine eimeriosis have morphologically similar oocysts that are difficult to differentiate. For the identification of particular Eimeria spp., diagnostic laboratories are increasingly turning to DNA-based technology. This study aims to develop a multiplex polymerase chain reaction (mPCR) technique based on the internal transcribed spacer-1 (ITS-1) gene for the simultaneous identification of pathogenic Eimeria spp. in cattle from Sulawesi Island, Indonesia. Materials and Methods: Genomic DNA was extracted by the DNAzol reagent from the purified Eimeria oocysts. Species-specific primers targeting the ITS-1 region were used to amplify the distinct Eimeria spp. Results: Using PCR ITS-1, this study showed that 36 of 120 fecal samples (30%) were infected by Eimeria spp. The multiplex PCR assay allowed for the simultaneous identification of six major Eimeria spp. in a single-tube reaction. The proportion of mixed Eimeria spp. infections was 100% (36/36). The maximum number of Eimeria spp. was five, and the minimum number was two. Conclusion: Identification of six pathogenic Eimeria spp. in cattle was successfully carried out by nested multiplex PCR using ITS-1 gene. In the future, a procedure to detect pathogenic Eimeria spp. in one tube reaction will offer economical and save diagnostic time.

<p class="MDPI17abstract"><strong>Objective: </strong>Kebumen Ongole Grade (Kebumen OG) are known as good performance cattle with body weight and body measurement higher than National Standard of Ongole Grade cattle. Productivity is influenced by genetic and environmental factors. <em>Insulin-induced gene-1</em> (<em>INSIG1</em>) gene is one of many genes that are considered important in influencing carcass characteristics and playing an important role in lipid metabolism (adipogenesis and or lipogenesis). So, the aim of this study was to identify of SNP A4366G in <em>INSIG1</em> gene and associated with slaughter characteristics of unproductive Kebumen OG cows.</p><p class="MDPI17abstract"><strong>Methods: </strong>In this study used 44 unproductive Kebumen OG cows from slaughter house in Kebumen. Slaughter characteristics contain of slaughter weight, hot carcass weight and dressing percentage. Three milliliters of blood samples were collected from <em>vena jugularis</em>. DNA were extracted from blood samples using salting out method. Genotyping of <em>INSIG1</em> gene (A4366G) used PCR-RFLP method with <em>Taq</em>I restriction enzyme. Genetic diversity data in this study were allele and genotype frequencies, heterozygosity, PIC and HWE. Association of genotypes of <em>INSIG1</em> gene with slaughter characteristics were analyzed using ANOVA univariate model.<strong></strong></p><p class="MDPI17abstract"><strong>Results: </strong>Polymorphic of <em>INSIG1</em> (A4366G) gene was found in the unproductive Kebumen OG cows. Three variants of genotypes (AA, AG, GG) with two alleles (A and G) were found with allele frequencies 0.795 for G allele. The population was in equilibrium genetic. Association of slaughter characteristics with genotypes were not significant (p&gt;0.05).<strong></strong></p><p><strong>Conclusions: </strong>Polymorphic of <em>INSIG1</em> gene (A4366G) was found in unproductive Kebumen OG cows population with dominant of G allele. The population was in genetic equilibrium. The association of slaughter characteristics with genotype of <em>INSIG1</em> gene (SNP A4366G) was not significant.</p>

<p class="MDPI17abstract"><strong>Objective: </strong>Bovine Leukocyte Adhesion Deficiency (BLAD) is a genetic disorder in Holstein-Friesian (HF) cattle that have a significant economic impact on dairy cattle breeding. Mutation c.383A&gt;G in Cluster of Differentiation Molecule 18 (CD18) gene was known as related to BLAD. This study aimed to detect the CD18 gene mutation that causes BLAD of HF cattle in the West Java Province.</p><p class="MDPI17abstract"><strong>Methods: </strong>The genomic DNA was extracted from 88 blood samples of HF cattle from Cibungbulang-Bogor (n=34), Ciampea-Bogor (n=31), and Sukabumi (n=23) by the GB300 DNA extraction kit, Geneaid^TM. The CD18 gene mutation was detected by PCR-RFLP analysis using the <em>Taq</em>I restriction endonuclease.<strong></strong></p><p class="MDPI17abstract"><strong>Results: </strong>All samples in this study produced two fragments DNA, i.e., 359 bp and 260 bp with the monomorphic homozygote genotype (BB). HF cattle with BB genotype indicated as normal cattle and did not carry BLAD.<strong></strong></p><strong>Conclusions: </strong>All samples in the three populations of West Java were free of BLAD genetic disorder. Identification of BLAD on HF cattle in other places, including bulls in the Center of Semen Production in Indonesia was needed to prevent this genetic disorder.