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American foulbrood is an important bacterial disease affecting the larvae of honey bees (Apis mellifera L.) caused by Paenibacillus larvae. Due to easy transmission of disease and the ability of bacteria to create spores, it is a bacterium resistant to both physical and chemical conditions. The study aims to isolate, perform microbiological analyses, and determine biochemical properties and genotypes P. larvae strains from AFB samples collected from Turkey's Central and Eastern Black Sea regions. An isolation study was conducted on adult bees, larvae, honey, and primary honeycomb samples from suspected colonies in the regions under study. After the purification of bacterial isolates from samples, P. larvae strains were identified using biochemical and molecular methods. The genetic diversity and ERIC types of P. larvae isolates were determined by rep-PCR DNA genotyping using BOX A1R and MBO REP1 primers and multiplex-PCR. A phylogenetic tree of P. larvae strains was constructed in the study. All P. larvae isolates were determined as ERIC I type. According to the rep-PCR results of P. larvae strains, 15 of the 28 isolates were Ab genotype (54%), 7 (25%) Aβ genotype, 4 (14%) AB genotype, 1 (3.5%) αB genotype, and 1 (3.5%) ab genotype. From an epidemiological viewpoint, it was determined that Ab and Aβ genotypes were widely distributed, while other genotypes (AB, αB, and ab) showed less spread. The results of the study will guide researchers in taking relevant measures to prevent and control American foulbrood. [ABSTRACT FROM AUTHOR]

Acarapis woodi (Rennie 1921) (Acari: Tarsonemidae) is one of the mites that settles in the respiratory system of honeybees (Apis mellifera L. (Hymenoptera, Apidae)) and distributed throughout the world. It causes significant economic losses on honey production. In Türkiye, studies on the existence of A. woodi are very limited and so far, no studies on the molecular diagnosis and phylogenetic of it have been reported in Türkiye. This study was conducted to investigate the prevalence of A. woodi in Türkiye, especially in areas where beekeeping is intense. Diagnosis of A. woodi was performed using both microscopic and molecular methods using specific PCR primers. Adult honeybee samples were collected from 1.193 hives in 40 provinces of Türkiye between 2018 and 2019. Based on identification studies, the presence of A. woodi was detected in a total of 3 hives (0.5%) in 2018 and 4 hives (0.7%) in 2019. This is the first report for determination of A. woodi in Türkiye. [ABSTRACT FROM AUTHOR]

Ascosphaera apis (Maassen ex Claussen) Olive et Spiltoir is a causal agent of chalkbrood disease and is widely found in mainly honeybee colonies worldwide. Chalkbrood disease rarely causes colony death, but severe infections can lead to a decrease in honeybee populations and as a result, a decrease in honey production or beekeeping byproducts. Therefore, it is important to determine the distribution and prevalence of this disease agent in a certain region in terms of controlling of it. In this study, it was aimed to identify chalkbrood disease agents affecting honeybee colonies especially in beekeeping places in Turkey, to isolate and identify them using morphological and molecular methods, especially ITS gene sequencing. Morphologically, pure cultures of clinically suspected samples in terms of chalkbrood disease were created and microscopically evaluated. After that, genomic DNA isolation was performed from pure cultures to amplify ITS gene region and the resulting gene sequences were used in molecular identification and phylogenetic analysis. For this purpose, a total of 1.193 hives were randomly selected from 400 apiaries in 40 provinces throughout Turkey between 2018 and 2019. As a result, the presence of A. apis was detected in 7 hives in 2018 and 19 hives in 2019, as a result a total of 26 samples were found to be positive. Accordingly, the infection rate was determined as 2.18%. The obtained results are thought to be important in terms of determining the spread rate of A. apis and the rate of chalkbrood disease in Turkey. [ABSTRACT FROM AUTHOR]

Paenibacillus larvae (P. larvae) is a pathogenic bacterium causing American Foulbrood Disease (AFB) in honeybee larvae. It is necessary to develop alternative strategies for the control of this disease due to the serious honeybee colonies loses and the use of antibiotics. Recent studies are aimed at the investigating natural fighting agents against P. larvae. In our study, it was aimed to demonstrate potential antibacterial efficacy of ethanol extract of Anatolian Propolis (EAP) against P. larvae strains PB35 and SV35 which were isolated in Turkey. The total phenolic content (TPC) and flavonoid content (TFC) of EEAP were determined as 181.73±5.20 mg Gallic Acid Equivalents (GAE)/g, and 42.33±1.40 mg Quercetin Equivalents (QE)/g, respectively. It was found that EAP contains different amounts of ferulic, caffeic, coumaric acids, rutin, and caffeic acid phenethyl ester (CAPE). The antibacterial activity of the EAP was determined by using agar-well diffusion, microdilution, and Bioscreen C techniques. The Minimal Inhibition Concentration (MIC) values of the EAP were determined as 74.87 μg/ml against strain PB35 and SV35 using both microdilution and Bioscreen C teqnique. In both techniques, Minimal Bactericidal Concentration (MBC) values of the EAP were evaluated as 149 and 598.4 μg/ml against strain PB35 and SV35, respectively. The fact that EAP shows low concentrations of bacteriostatic (MIC) and bactericide (MBC) activity values against P. larvae strains, spore-forming bacilli, which are agents of AFB disease, suggests that it may be a potential source in AFB disease control. [ABSTRACT FROM AUTHOR]

● The new phage Paenibacillus phage SV21(vB_PlaP_SV21) was isolated, characterized, and compared to other known paenibacilus phages. ● The whole-genome analysis determined no known tRNA or antibiotic resistance genes. ● Paenibacillus phage SV21(vB_PlaP_SV21) is a new virulent phage belonging to the podoviridae family, which can infect theirs specifically the Paenibacillus larvae causative of AFB disease. Paenibacillus larvae cause an American foulbrood disease (AFB) that is responsible for the extinction of honeybee colonies and is a honeybee bacterial disease that has to be obligatory notified worldwide. Recently, bacteriophage studies targeting Paenibacillus larvae have emerged as a promising alternative treatment method. The inability of bacteria to create resistance against bacteriophages makes this method advantageous. As a consequence, this study was conducted to describe the genome and biological characteristics of a novel phage capable of lysing Paenibacillus larvae samples isolated from honeybee larva samples in Turkey. The Paenibacillus phage SV21 (vB_PlaP_SV21) was isolated by inducing Paenibacillus larvae strain SV21 with Mitomycin-C. Whole-genome sequencing, comparative genomics, and phylogenetic analysis of vB_PlaP_SV21 were performed. Transmission electron microscopy images showed that vB_PlaP_SV21 phage was a Podovirus morphology. The vB_PlaP_SV21 phage specific for Paenibacillus larvae was determined to belong to the Podoviridae family. Host range and specificity, burst size, lytic activity, and morphological characteristics of the phage were determined. Bioinformatic analysis of the Paenibacillus phage SV21 showed 77 coding sequences in its linear 44,949 bp dsDNA genome with a GC content of 39.33%. In this study, we analysed the genomes of all of the currently sequenced P. larvae phage genomes and classified them into five clusters and a singleton. According to molecular, morphological, and bioinformatics results, ıt was observed that API480 (podovirus), which was reported as a singleton in previous studies and public databases, and Paenibacillus phage SV21 phage could form a new cluster together [ABSTRACT FROM AUTHOR]

Purpose: Bumblebees are an important group of insects in the pollination of various vegetables, fruits, oilseeds, legumes, and the fodder crops. Compared to honeybees, they have a wider choice of hosts and a longer flight period. These bees are used especially for the pollination of plants in greenhouses and are commercially produced for this purpose. Recently, serious decreases have been occurring in bumblebee populations due to various reasons such as pathogens, and some of species are even threatened with extinction. Due to the worldwide decline in pollinator insects, determining the distribution and prevalence of bumblebee pathogens is of great importance. Therefore, this study was conducted to determine the incidence and prevalence of pathogens in Turkish bumblebee populations and how much of each pathogen was in bumblebee samples. Methods: A total of 172 Bombus terrestris (Linnaeus,1758) samples (21 samples from commercial enterprises, 79 samples from greenhouses and 72 samples from nature) were randomly collected from 3 provinces (Antalya, Mersin and İzmir) where greenhouse cultivation is intensively carried out in Turkey. Eighty-nine of these samples were collected in the spring and eighty-three in the autumn. The presence of four pathogens (Nosema bombi, Crithidia bombi, Apicystis bombi, and Locustacarus buchneri) was investigated by PCR using universal primers. Results: The overall prevalence of Nosema bombi, Crithidia bombi, Apicystis bombi, and Locustacarus buchneri was determined as 7.55%, 9.3%, 11.62%, and 4.65%, respectively. Co-infections (5.81%) were only detected in wild-caught (nature) samples. C. bombi and A. bombi infections were detected at higher rates in the spring samples than in the autumn samples (p < 0.05). There was no significant difference between the spring and autumn samples with respect to the presence of N. bombi and L. buchneri (p > 0.05). Conclusion: The results obtained could be important in determining the prevalence and spread rates of the bumblebee diseases in Turkey and to determine appropriate protection measures. The information gathered should increase our knowledge about the presence of these pathogens in Turkey and could contribute to improve apiarist's practice. More studies are needed to determine the transmission pathways of these pathogens between the populations. Also, complex pathogen interactions in bumblebee populations should be considered in the future to improve bumblebee health. [ABSTRACT FROM AUTHOR]

The aim of this study was to investigate the chemical composition of the essential oils (EOs) from two Diplophyllum species and to evaluate their bioactivity potential [antimicrobial, antioxidant, anti-urease, anti-xanthine oxidase (XO)], and phenolic compounds. The analysis of Diplophyllum albicans and Diplophyllum taxifolium permitted the identification of 62 components, comprising = 99.6% of the total EO composition. The major components found in these liverwort species were ß-patchoulene, ß-santalene, ß-himachalene, and cubebol. The antimicrobial assays showed that the solvent extracts (n-hexane and methanol) from these liverwort species exhibited weak to moderate antimicrobial activity. In addition, the methanol extracts of these liverwort species also exhibited moderate to high antioxidant potential. The enzyme inhibitory effects of the species were determined using urease and XO for the methanol extracts. Generally, the methanol extracts of D. albicans and D. taxifolium exhibited powerful urease inhibition with IC50 values of 9.711 ± 0.058 and 6.304 ± 0.099 µg/mL, respectively, compared to the IC50 value of 26.124 ± 0.077 µg/mL for the standard (acetohydroxamic acid). Moreover, the HPLC-UV detection method showed that the analysed methanol extracts of these liverwort species contained only catechin and benzoic acid. These findings suggest that the analysed liverwort species possess antioxidant and urease inhibition, thus indicating the potential to explore new bioactive molecules. Cilj ove studije bio je da se ispita hemijski sastav eteričnih ulja (EO) dve vrste roda Diplophyllum i proceni potencijal bioaktivnosti [antimikrobna, antioksidantna, antiureazna, antiksantin oksidaza (KSO)] i fenolna jedinjenja. Kod Diplophyllum albicans i Diplophyllum taxifolium identifikovane su 62 komponente, koje čine = 99,6% ukupnog sastava EO. Glavne komponente nađene u ovim vrstama jetrenjača su ß-pačulen, ß-santalen, ß-himahalen, i kubebol. Antimikrobni testovi su pokazali da ekstrakti rastvarača (n-heksan i metanol) iz ovih vrsta jetrenjača pokazuju slabu do umerenu antimikrobnu aktivnost. Pored toga, metanolni ekstrakti ovih vrsta pokazali su umeren do visok antioksidativni potencijal. Enzimski inhibitorni efekti vrsta su određeni korišc'enjem ureaze i KSO za metanolne ekstrakte. Generalno, metanolni ekstrakti D. albicans i D. taxifolium su pokazali snažnu inhibiciju ureaze sa IC50 vrednostima 9,711 ± 0,058 i 6,304 ± 0,099 µg/mL, respektivno, gde je IC50 standarda (acetohidroksamične kiseline) 26,124 ± 0,077 µg/mL. Pored toga, HPLC-UV detekcija je pokazala da analizirani metanolni ekstrakti ovih vrsta jetrenjača sadrže samo katehin i benzoevu kiselinu. Ovi rezultati pokazuju da analizirane vrste jetrenjača poseduju antioksidantnu aktivnost i inhibiciju ureaze; te stoga poseduju potencijal za istraživanje novih bioaktivnih molekula. [ABSTRACT FROM AUTHOR]

Simple Summary: The health of bees is suspected to be low in Turkey due to the lower amounts of bee products that they produce per colony. Regular monitoring of bee diseases in Turkey has been lacking. We sampled bees from 115 colonies across five different locations to determine which pathogens are present and how much of each pathogen was in a bee colony. We found that the Varroa mite is widespread and consistently found in 90% of the bee colonies. We also found that pathogens normally transmitted by this parasite was also present in nearly 100% of the colonies sampled. We therefore concluded that the presence of Varroa mites is central to the decline in bee health and that management practices targeted for this parasite will most likely improve bee health in Turkey. In addition, three bee viruses are interacting with one another that influence the susceptibility to a more deadly variant responsible for colony death. Therefore, these viral interactions should be considered in the future to devise effective ways to improve honey bee health. Beekeeping has yet to reach its full potential in terms of productivity in Turkey where it has a relatively large role in the economy. Poor colony health is suspected to be the reason for this, but comprehensive disease monitoring programs are lacking to support this notion. We sampled a total of 115 colonies across five different apiaries throughout the Marmara region of Turkey and screened for all of the major bee pathogens using PCR and RNA-seq methods. We found that Varroa mites are more prevalent in comparison to Nosema infections. The pathogens ABPV, DWV, KV, and VDV1 are near 100% prevalent and are the most abundant across all locations, which are known to be vectored by the Varroa mite. We therefore suspect that controlling Varroa mites will be key for improving bee health in Turkey moving forward. We also documented significant interactions between DWV, KV, and VDV1, which may explain how the more virulent strain of the virus becomes abundant. ABPV had a positive interaction with VDV1, thereby possibly facilitating this more virulent viral strain, but a negative interaction with Nosema ceranae. Therefore, these complex pathogen interactions should be taken into consideration in the future to improve bee health. [ABSTRACT FROM AUTHOR]

Nitrate and nitrite ions taken from food are the sources of bioavailable nitric oxide (NO) in the nitrogen cycle. Some beneficial effects of honey on health are attributed to the ability of honey to increase NO production. The variation of nitrate and nitrite levels of honey samples collected from different Anatolia regions were clarified using capillary electrophoresis technique. The sensitivities of both anions were improved with the application of the sample stacking method. Separation buffer consisted of 30 mmol L−1 formic acid and 30 mmol L−1 sodium sulfate at a pH of 4.0. The CE technique revealed that 18 honey samples contained nitrate anion ranged between 2.53 and 31.8 mg kg−1. Nitrite amounts were found in lower amounts in the honey samples as between non-detected and 0.533 mg kg−1. The observed differences in nitrate levels between honey varieties may be a way to determine honey's origin. [ABSTRACT FROM AUTHOR]

The aim of this study is to analyze the volatile composition and antimicrobial and antioxidant activities of the essential oils of Polytrichum commune and Antitrichia curtipendula. The essential oils obtained by hydrodistillation (HD) from each species were identified by GC-MS/FID. The main components were biformene (13.06%), α-pinene (6.53%), and bornyl acetate (8.10%) in P. commune. Nonanal and tetradecanal as major compounds were 19.96% and 20.23% in A. curtipendula essential oils, respectively. Antioxidant activity of obtained essential oils was evaluated using in-vitro antioxidant models. There was no significant difference within the groups according to DPPH activity. Also, the essential oil from P. commune showed higher metal-ion chelating activities than that of the essential oil of A. curtipendula. Metal-ion chelating activities varied between 4.1% and 67.4% at the 800 μg/mL concentration, respectively. The antimicrobial activity was tested by a minimal inhibition concentration test. Each moss species showed good antimicrobial activity against microorganisms according to the results of minimal inhibition concentration experiments. [ABSTRACT FROM AUTHOR]

In this study, volatile compounds (VCs) in the essential oil (EO), SPME and SPME of n-hexane extract of Vinca major subsp. hirsuta (Boiss.) Stearn were analyzed by GC-FID/MS instrument. A total of 32, 38, and 26 compounds with in the 98.5%, 98.6%, and 98.6% were identified, respectively. As a result of VCs study, (Z)-3-hexenol (36.8%) in the EO, 1,3,5-trimethylbenzene (31,5%) in the SPME, and phenylethyl alcohol (32.8 %) in the SPME of n-hexane extract of V. major subsp. hirsuta were found to be major compounds. Sesquiterpenes (20.6%, and 11.5%) for the HD and SPME, monoterpenes (18.6%) for the SPME of n-hexane extract of V. major subsp. hirsuta were found as the main groups among the terpenic compounds, respectively. Then, the antimicrobial activity of the EO and the solvent extracts (n-hexane, acetonitrile, methanol, and water) of V. major subs. hirsuta against 3 gram negative, 3 gram positive, 1 tuberculosis and 2 fungus were screened. The EO showed the only activity against the Mycobacterium smegmatis ATCC607 (MIC, 152 µg/mL) and Candida albicans ATCC60193 (MIC, 38 µg/mL). The n-hexane extract did not show any activity against all tested microorganisms. The best antimicrobial activity for the acetonitrile, methanol, and water extracts were observed against M. smegmatis with 69 µg/mL, 609 µg/mL, and 437 µg/mL MIC values, respectively. None of the extracts were found to be active to Enterococcus faecalis ATCC29212, Staphylococcus aureus ATCC25923, and Saccharomyces cerevisiae RSKK 251. [ABSTRACT FROM AUTHOR]

Lamiaceae, a family of flowering plants, have been historically used as culinary herbs, shrubs and medicinal plants. With this study, the volatile organic compounds (VOCs) of the five species of the Lamiaceae family (Thymus cilicicus, Clinopodium nepeta L., Nepeta italica L., Thymbra spicata L. and Ziziphora clinopodioides) that grow in Turkey were identified by the SPME/GC‐FID/MS method. The five species were screened in order to reveal their antibacterial and antioxidant activities. Phenolic compound analyses of five species were performed by using RP‐HPLC with diode array detection. Among the total number of 73 compounds that were identified, α‐pinene (2.2%‐13.8%), β‐pinene (0.3%‐25.9%), p‐cymene (0.7%‐17.9%), γ‐terpinene (0.4%‐22.0%), carvacrol (0.1%‐40.2%), sabinene (1.2%‐34.2%), terpinen‐4‐ol (0.2%‐27.5%), (E)‐β‐caryophyllene (0.1%‐13.8%) and (E)‐β‐farnesene (0.4%‐19.7%) were found to be the most plentiful ones. The minimal inhibitory concentration (MIC) values of the methanol extracts of the five species of the Lamiaceae family were investigated, and the results were in the range of 23‐959 µg/mL; the most pronounced antibacterial activities were determined to be against Mycobacterium smegmatis (23‐291 µg/mL). All of the methanol extracts possessed effective antioxidant capacity (IC50: 1.82‐28.18 μmol/L Trolox/g sample) as estimated from the 2,2‐diphenyl‐1‐picrylhydrazyl (DPPH) radical‐scavenging assays. Additionally, the effect of phenolics, especially benzoic acid (86.27, 27.85, ‐, 13.11, 9.59 mg/g extract, respectively), on the biological properties of the methanol extracts of the five species of the Lamiaceae family, was investigated. [ABSTRACT FROM AUTHOR]

The chemical composition of the essential oils from all parts of Pseudoscleropodium purum, Eurhynchium striatum and Eurhynchium angustirete were analysed by GC-FID-MS. Sixty-five, thirty-four and seven compounds, accounting for 99.7%, 97.3% and 99.9% of the oils, were identified and the main components were α-pinene (16.1%), 3-octanone (48.1%), and eicosane (28.6%), respectively. The essential oils were also tested against nine strains using a broth microdilution method and showed moderate antimicrobial activity with minimum inhibitory concentrations (MIC) ranging from 278.2 to 2225 μg/mL. All the mosses essential oils showed good antituberculosis activity against Mycobacterium smegmatis with MIC of 278.2-312.0 μg/mL. [ABSTRACT FROM AUTHOR]