Bacterial diversity of loggerhead and green turtle eggs from two major nesting beaches from the Turkish coast of the Mediterranean.

This study was conducted during the 2018 nesting season at the Sugözü Beaches (Adana-Turkey) and Göksu Delta (Mersin-Turkey). Egg samples (n = 63) from loggerhead and green turtle nests (n = 43) were collected. Isolated bacteria were initially identified by phenotypic methods and then by MALDI-TOF MS. The bacterial mass spectra were analyzed using Principal Component Analysis. Bacterial isolation was performed for 55 isolates belonging to 12 genera from two major nesting sites. In Sugözü Beaches 62.2% of the bacteria species belonged to Enterobacteriaceae and in Göksu Delta 44.4% of the bacteria species belonged to Morganellaceae. Klebsiella oxytoca and Staphylococcus haemolyticus had not previously been detected in any sea turtle nests. This is the first MALDI-TOF MS study conducted for determination of bacterial variability in loggerhead turtle eggs in Turkey and serves as a reference study for the assessment of bacterial threat in sea turtle nests, enabling the establishment of suitable conservation measures and treatment processes for both sea turtles and nesting sites.

The microbial community in a green turtle nesting beach in the Mediterranean: application of the Biolog EcoPlate approach for beach pollution.

This study aims to characterize the microbial community and its relationship with heavy metal pollution in the beaches of Sugözü, an important nesting site for the green turtle. Heavy metal concentrations of sand samples from subregions of Sugözü were determined using ICP-MS. The microbial community was analyzed using the Biolog® EcoPlate. The relationship between microbial catalytic activity and heavy metal levels were analyzed using canonical correspondence analysis. Levels of 27Al, 57Fe, 55Mn, and 52Cr were quite high (4332.34, 13,764.77, 590.98, and 48.21 mg/kg, respectively). The microbial community in subregions with high levels of metals was found to use carboxylic acid as a carbon source. Bioactivity, substrate utilization, diversity, and evenness values indicated negative correlations concentrations of 27Al, 56Fe, and 52Cr (-0.820, -0.508, and -0.560, respectively). It was also found that microbial diversity decreased in the subregions where heavy metal concentration increased. Embryonic deaths were found highest at early stage (0.1 to 0.2 eggs) and lowest at middle stage for whole study sites by inspecting a total 6408 eggs of 63 green turtle nests. The Biolog EcoPlate was firstly applied to determine pollution, and our findings clearly demonstrate the applicability and effectiveness of this method in assessing nesting beaches.

Bacterial diversity of the green turtle (Chelonia mydas) nest environment.

The green turtle is an endangered species that is highly sensitive to environmental pollution that can adversely affect the healthy development of eggs. Moreover, the presence of some bacteria in nests can be regarded as an indicator of the pollution level in nesting areas. In our study, nest sand and egg contents were collected from Sugözü Beaches (Turkey), in the Mediterranean. Phenotypic and genotypic identification of bacteria were carried out by using conventional phenotypic methods, 16S rRNA gene sequencing respectively. The extended-spectrum beta-lactamase presence and carbapenem resistance of bacteria isolated from egg contents were determined. This is the first report of carbapenem resistance in the eggs. All strains were evaluated in three different categories including growth promoters in agriculture and aquaculture, pathogens that are found in human and animal, and biomonitoring aquatic pollution. According to our analysis, 67 bacterial species were identified from samples. This study is the first record of Alcaligenes, Zobellella, Lysinibacillus, Sphingobacterium, Achromobacter, Acinetobacter, Alcanivorax, Ochrobactrum, Microbacterium, Rhodococcus, and Stenotrophomonas isolated from sea turtles. Pathogens detected in the bacterial flora can threaten both sea turtles and field workers. These data can contribute to the development of new conservation strategies on the treatment of sea turtles, nest protection, and pollution detection on nesting beaches.

Molecular identification of fungal isolates and hatching success of green turtle (Chelonia mydas) nests.

The aim of this study is to investigate the fungal diversity of green turtle nests and to examine phylogenetic relationships among these isolates. During the nesting season, samples of intra-nest sand and failed eggs were collected from 25% of the surviving nests in Sugözü Beaches, which are amongst the most important nesting beaches for endangered green turtles in the Mediterranean. Twenty-three fungi were identified by molecular techniques. Fungal isolates belonged to genera Aspergillus, Emericella, Rhizopus, Actinomucor and Apophysomyces with two undescribed species. Aspergillus variecolor, Aspergillus quadrilinieatus, Aspergillus tubingensis, Rhizopus oryzae, Actinomucor elegans and Apophysomyces variabilis were firstly detected in all sea turtle nests within this study. Our results demonstrate that 36.4% of the nests had fungal contamination. Also hatching success of the nests contaminated by fungi were significantly lower than the uncontaminated nests (P = 0.029). Also, this may represent a threat to marine turtles and a risk for the health of conservation workers. This study is the first molecular phylogenetic study associated with sea turtle nests in the eastern Mediterranean coast and contributes to the wider body of literature on fungal invasion of sea turtle nests with firstly isolated species. These findings are important for improving potential conservation measures for the nest sites.

Escherichia Coli: Characteristics of Carbapenem Resistance and Virulence Factors

ABSTRACT In this study, fifty Escherichia coli strains were analyzed by multiplex polymerase chain reaction for the genes expressed carbapenemase and virulence factors in order to determine the presence of carbapenemase and nine virulence factors and investigate the association between these two characteristics. When carbapenemase susceptibility was taken into consideration, OXA-48 type carbapenemase was determined for 22% of the total strains. Also, the frequency of virulence gene regions in E.coli infections and virulence gene profiles of these isolates were examined and the frequency of pap, afa, sfa, fimA, iroN, aer, iutA, hly and cnf-1 genes were 24, 38, 20, 84, 28, 90, 92, 10 and 34% respectively. A significant correlation was found between the presence of fimA and afa gene regions and carbapenem susceptibility (P< 0.05). Based on the combination of carbapenemase and virulence factor genes, 24 different gene profiles were determined for all strains. The results of the study appear to indicate that fimA and afa genes correlate with carbapenem susceptibility, the relations of fimA with urinary tract infections and pap with complicated urinary tract infections. It also indicates that sfa and afa genes correlate with other infections except urinary tract infections.

Klebsiella pneumoniae: characteristics of carbapenem resistance and virulence factors.

Klebsiella pneumoniae, known as a major threat to public health, is the most common factor of nosocomial and community acquired infections. In this study, 50 K. pneumoniae clinical specimens isolated from bronchial, urea, blood, catheter, rectal, bile, tracheal and wound cultures were collected. These isolates were identified and carbapenem resistance was determined via an automated system, CHROMagar Orientation and CHROMagar KPC. The carbapenemase gene regions (blaIMP, blaVIM, blaOXA, blaNDM and blaKPC) and presence of virulence factors (magA, k2A, rmpA, wabG, uge, allS, entB, ycfM, kpn, wcaG, fimH, mrkD, iutA, iroN, hly ve cnf-1) of these isolates were determined by using Multiplex-PCR. The OXA-48 carbapenemase gene regions were determined in 33 of 50 K. pneumoniae strains. In addition, NDM-1 resistance in one, OXA-48 and NDM-1 resistance in four unusual K. pneumoniae isolates were detected. Virulence gene regions that were encountered among K. pneumoniae isolates were 88% wabG, 86% uge, 80% ycfM and 72% entB, related with capsule, capsule lipoprotein and external membrane protein, responsible for enterobactin production, respectively. Even though there was no significant difference between resistant and sensitive strains due to the virulence gene regions (P≥0.05), virulence factors in carbapenem resistant isolates were found to be more diverse. This study is important for both, to prevent the spread of carbapenem resistant infections and to plan for developing effective treatments. Moreover, this study is the first detailed study of the carbapenem resistance and virulence factors in K. pneumoniae strains.