Evaluation of the occurrence of genetic determinants of multi-drug resistance among Acinetobacter baumannii strains

Introduction: The aim of the study was the analysis of occurrence of genetic determinants of multi-drug resistance and the assessment of genetic relationship among Acinetobacter baumannii strains. Methods: Multiplex-PCR method was performed in order to: (1) confirm the phenotypic identification and (2) detect the presence of CHDL oxacillinases in the group of thirty A.baumannii strains. Further PCR studies included the analysis of the occurrence of genetic determinants associated with efflux pump, insertion sequence and biofilm formation. The relationship between bacterial strains was assayed using 6 primers in RAPD-PCR method. Results: Detection of the blaOXA-51-like gene confirmed that the strains belong to the A. baumannii species. In the multiplex-PCR, the presence of the blaOXA-23-like and blaOXA-40-like genes was detected in 3 (10%) and 27 (90%) isolates, respectively. Moreover, some strains showed the coexistence of the blaOXA-51-like and blaOXA-23-like genes (10%, n=3) or blaOXA-51-like and blaOXA-40-like (90%, n=27). In the group of analysed strains the presence of the efflux pump gene (adeA) and the insertion sequence ISAba1 were demonstrated in all tested isolates. Biofilm-related genes (abaI, csuE) were found in 100% and 97% (n=29) tested strains adequately. The RAPD-PCR studies revealed the presence of 10 unrelated genotypes. Conclusions: The obtained results suggest that the phenomenon of multi-drug resistance in the studied A. baumannii strains could be attributed to the occurrence of CHDL oxacillinases, AdeABC efflux pump, insertion sequence ISAba1 and the biofilm formation.

Genetic Distance of Four Strains of Guppy Fish (Poecilia reticulata) Using RAPD PCR Method

Guppies are one type of organism that can live well, especially in the tropics. The kinship between species that have genetic similarities can be identified through genotypic mapping, one of which is molecular analysis using. RAPD method with PCR technique. This study aims to determine the genetic relationship of four guppy poecilia reticulata strains, respectively are Albino Full Red (AFP), Brazilian Fan Tail (BFT), Koi Guppy Tuxedo (KGT), and Platinum Red Tail Big Ears (PRTB) with the RAPD-PCR method. The genetic relationship data obtained is used as a guide for mating between the four strains. This research was conducted from September 2020 to November 2020. The process were carried out at the Biotechnology Laboratory, Faculty of Fisheries and Marine Sciences, Padjadjaran University and the Central Laboratory, Padjadjaran University. OPA-03 primer (AGTCAGCCAC) is used as a standard parameter to interpret genetic diversity among the four guppy strains. Based on the results, amplification with primer OPA-03 visualized 21 bands consisting of seven polymorphic bands and 13 monomorphic bands. The results of the phylogenetic tree showed that there were two groups. The first group is AFR and BFT a similarity index of 69.5%. The second group is KGT and PRTB a similarity index of 71.5%.

Genetic Diversity of Orobanche cumana Populations in Serbia

In this study, we report genetic characterization of Orobanche cumana, the causal agent of sunflower wilting in Serbia. The genetic diversity of this parasitic plant in Serbia was not studied before. Random amplified polymorphic DNA (RAPD) markers and partial rbcL gene sequences analysis were used to characterize the O. cumana populations at the molecular level. While phylogenetic analyses of RAPD-PCR amplicons were performed using unweighted pair-group Method analyses, rbcL gene sequences were analyzed using neigbor joining method and minimum spanning tree. Molecular analyses of RAPD-PCR analysis revealed high genetic diversity of O. cumana populations which indicated high adaptive potential of this parasitic weed in Serbia. Further analyses of rbcL gene using minimum spanning tree revealed clear differences among diverse sections of Orobanche genus. Although this molecular marker lacked the resolution to display intrapopulation diversity it could be a useful tool for understanding the evolution of this parasitic plant. Our results suggested that O. cumana has great genetic potential which can lead to differentiation of more virulent races which is important for determining crop breeding strategies for their control.

Molecular Classification of Vicia faba L Genotypes by Using RAPD-PCR Markers

The research included the molecular classification study of seven genotypes of the bean Vicia faba L. (FBSPN2, TLD1266, TLD1814, TLB1266, Luzdeotono, favad and Histal. Using the RAPD technique for DNA, as 13 random primers were used, the products of inflation were transferred within the agarose gel, and the results of the study showed the possibility of separating the genotypes from each other and determining the degree of genetic variation between them, as the primers used produced (1002) packages of them (417 normal bundles and (585) mixed bundles. The genetic differences of the studied genotypes were determined to be distinguished by the number of bundles, as they reached (28) bundles, including (13) unique bundles and (15) absent bundles. The ILB1266 genotype showed the highest number of unique bundles, which It reached 4 bundles, while the cultivar Favad showed the absence of unique bundles in it, either bundles are absent. The genotypes (ILD1266, IILB1266, Luzdeotono) were distinguished for having the highest number, which amounted to (3) bundles. As for the FBSPN2 genotype, it did not have any absent bundle, and the primers varied. Of the resulting bundle sizes, their sizes ranged between bp (1925-130), and the highest value for the genetic dimension ranged between (0.110 - 0.269), as the lowest genetic dimension was between the two structures (FBSPN2 and ILD1266), which amounted to 0.110, and the highest value for the genetic dimension was (0.2 69) between the genotypes (ILD1266, HISTAL) (ILD1266, Luzdeotono) The Dendrogram shows the separation of the studied genotypes into two main groups, and each of them into two subgroups.

ANALYSIS OF GENETIC DIVERSITY OF 26 CASSAVA VARIETIES (Manihot esculenta CRANTZ) WITH DIFFERENT RESPONSES TO WATERLOGGING CONDITIONS BY USING RAPD MARKERS

RAPD (Randomly Amplified Polymorphic DNA) is an indicator for high and stable polymorphism, widely used in the study of the diversity of cassava. In this paper, the results of using 20 polymorphic primers OPK combined with the establishment of the phylogenetic tree to analyze the genetic diversity of 26 cassava varieties with different responses to waterlogging conditions by using the RAPD-PCR technique were presented. The purpose of this experiment was to show the genetic relevance of the studied cassava varieties. The results showed that the flood tolerance of cassava was not related to the polymorphism and branching characteristics of the stem. This information may be use as a basis for selecting flood-tolerant cassava varieties for cassava production, as well as the basis for selecting genetically different parents for breeding.

Investigations on Transfer of Pathogens between Foster Cows and Calves during the Suckling Period

To date, there have been few studies on the health effects of foster cow systems, including the transmission of mastitis-associated pathogens during suckling. The present study aimed to compare the pathogens detected in the mammary glands of the foster cow with those in the oral cavities of the associated foster calves and to evaluate the resulting consequences for udder health, calf health and internal biosecurity. Quarter milk sampling of 99 foster cows from an organic dairy farm was conducted twice during the foster period. Oral cavity swabs were taken from 345 foster calves. Furthermore, quarter milk samples were collected from 124 biological dams to investigate possible transmission to the foster cows via the suckling calves. All samples were microbiologically examined and confirmed by MALDI-TOF (matrix-assisted laser desorption time-of-flight mass-spectrometry). Using RAPD-PCR (randomly amplified polymorphic DNA polymerase chain reaction), strain similarities were detected for Pasteurella multocida, Staphylococcus aureus, S. sciuri and Streptococcus (Sc.) suis. Transmission of P. multocida and S. aureus probably occurred during suckling. For S. sciuri and Sc. suis, environmental origins were assumed. Transmission from dam to foster cow with the suckling calf as vector could not be clearly demonstrated.