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Assessment of biocontrol potential of different Steinernema species and their bacterial symbionts, Xenorhabdus species against larvae of almond moth, Ephestia cautella (Walker)
(Pergamon-Elsevier Science Ltd, 2023) Yüksel, Ebubekir; Ormanoğlu, Nesrin; İmren, Mustafa; Canhilal, Ramazan
The almond moth, Ephestia cautella is a serious pest of stored food commodities including dried figs, dates, raisins, cocoa beans. The control of E. cautella infestations in storage that mostly relies on toxic insecticides or fumigants has caused emergence of resistant pest populations as well as residues on the commodities. Ento-mopathogenic nematodes (EPNs) and their endosymbionts are one of the emerging approaches in eco-friendly pest management and pose no health risk to humans and the environment. Therefore, here we evaluated the toxicity of different EPN species (Steinernema feltiae, S. carpocapsae, and S. bicornotum) and their endosymbiotic bacteria (Xenorhabdus nematophila, X. bovienii, and X. budapestensis) on the 3rd and 4th instars larvae of E. cautella under controlled conditions. The IJs of EPNs were applied to concrete arenas in Petri dishes at three concen-trations (50, 100, and 200 IJs/cm2). Oral and contact toxicity of the cell-free supernatants and cell suspension of symbiotic bacteria (4x107 cells/mL) were tested against both larval instars of E. cautella. Steinernema carpocapsae and S. feltiae induced mortalities over 80% at their lowest concentrations against both larval instars after 24 h of application. Oral and contact treatments of cell-free supernatants and cell suspensions were lethal for both larval instars; however, higher mortalities were generally obtained from direct exposure of larvae to cell-free super-natants and cell suspensions. The contact treatment of X. nematophila exhibited the highest virulence, causing 70 and 82.5% mortalities in 3rd and 4th instar larvae after 72 h of application, respectively. The anti-feedant ac-tivity of cell-free supernatants and cell suspensions was also observed in both instars in oral treatments. The findings of this study suggest that EPNs and their symbiotic bacteria have enormous potential for the eco-friendly control of E. cautella.
Assessment of the natural presence of entomopathogenic nematodes in Kayseri soils, Turkey
(Springer, 2017) Canhilal, Ramazan; Waeyenberge, Lieven; Yüksel, Ebubekir; Koca, Abdurrahman Sami; Deniz, Yunus; İmren, Mustafa
Entomopathogenic nematodes (EPNs) have lots of ideal attributes as biological control agents, such as a wide host range, rapid host mortality, ease of commercial production in vivo or in vitro, active host-seeking ability, long-term efficacy, easy application, compatible with many chemical pesticides and environmental safety. In the present study, 61 EPNs isolates from 174 soil samples were collected for the districts of Kayseri Province and from 5 different habitats (Forest, pasture, field crops, vegetable, and fruit orchards). The positive soil samples ratio was (35.1%). Forty-seven of isolates were Steinernema with the ratio of (27%). Prevalence of Heterorhabditis was (8%) (14 isolates). Nematodes were identified by morphometric and molecular methods. Forty-one of these isolates, were identified as Steinernema feltiae. Two isolates were S. carpocapsae, one was S. bicornutum, 6 were unknown Steinernema spp., and 14 were Heterorhabditis bacteriophora. Of these nematodes, S. bicornutum was a first record in Turkey. The soil textures of the EPN positive soils were loam, sandy loam, loamy sand and sandy clay loam with weakly basic (pH 7.57-7.91) and 1.15-3.06% organic content. The electrical conductivity of the nematode positive soils varied from 0.19 to 0.79 (none saline) dS/m.
Azadirachtin ve Spinosadın Bazı Yerel Entomoapatojen Nematod İzolatlarının Canlılığıve Virülensliği Üzerine Etkileri
(2019) Yüksel, Ebubekir; Imren, Mustafa; Canhilal, Ramazan
Entomopatojen nematodlar (EPN), biyoloijk mücadele uygulamaları içerisinde çoğu zaman biyopestisitlerle birlikte kullanılmaktadır. Biyopestisitlerin EPN’lar üzerindeki toksik etkilerinin belirlenmesi, bu uygulamalardan istenilen başarının elde edilebilmesi için oldukça önemlidir. Bu çalışmada yaygın birşekilde kullanılan Nimiks (40 g L-1 Azadirachtin) ve Laser (Spinosad 480 gL-1) biyopestisitlerinin bazı yerel EPN türlerinin (Steinernema feltiaeE-76, Heterorhabditis indica216-H, S. littoraleMGZ-4-S) hayatta kalma ve virülensliği üzerindeki etkileri 24 ve 48 saatlik periyotlarda son dönem Galleria mellonellalarvaları üzerinde laboratuvar koşullarında (25oC±2, R.H. %65±5) araştırılmıştır. Spinosad ve Azadirachtinin tavsiye edilen en yüksek uygulama dozlarına doğrudan maruz kalan EPN türleri içerisinde ilk 24 saatlik periyotta en tolerant türün %94 canlılık oranıyla H.indica216-H olduğu belirlenmiştir. Steinernema littoraleMGZ-4-S izolatının Azadirachtine 24 ve 48 saat periyotlarda sırasıyla %11 ve %12 ölüm oranlarıyla en çok duyarlılık gösteren tür olmuştur. Test edilen EPN türlerinin son dönem G. mellonellalarvaları üzerinde meydana getirdikleri ölüm oranları %80 ile %100 arasında değişmiştir. Elde edilen veriler, test edilen EPN türlerinin Spinosad ve Azadirachtin ile laboratuvar şartlarında uyumlu olduğunu göstermektedir.
Corrigendum to “Potential of four Turkish isolates of entomopathogenic nematodes against three major stored products insect pests” [J. Stored Prod. Res. 83 (September 2019) 317–321] (Journal of Stored Products Research (2019) 83 (317–321), (S0022474X19302826), (10.1016/j.jspr.2019.08.003))
(Elsevier, 2021) Yüksel, Ebubekir; Canhilal, Ramazan; İmren, Mustafa
The authors regret that the following statement was not added inadvertently to the footnote of the original manuscript when published online. < This article was produced from a master thesis carried out in 2017>. The authors would like to apologise for any inconvenience caused.
Determination of Resistance Levels of National Potato Cultivars and Clones Against Golden Cyst Nematode Pathotype Ro2/3 via Phenotypic and DNA Marker-Assisted Characterization
(Springer, 2024) Avci, Gulten Kacar; Canhilal, Ramazan; Toktay, Halil; Imren, Mustafa; Unlenen, Levent; Pirlak, Ugur
Potato (Solanum tuberosum L.) is one of our important agricultural products, which is the main food source for people in T & uuml;rkiye, as well as all over the world. There are many diseases and pests that reduce productivity in potato plant production. Potato cyst nematodes (Tylenchida: Heteroderidae) are pests that are on the quarantine list of the European and Mediterranean Plant Protection Organization and cause serious yield losses. Since they are soil-borne pathogens and there is no effective chemical control, the most successful control method is to use resistant cultivars. The aim of the study was to determine the resistance levels of local and national potato cultivars and clones developed by the Nigde Potato Research Institute against the Globodera rostochiensis Ro2/3 pathotype using molecular marker analysis and biotesting methods. The biotest study was carried out by inoculating 7500 eggs and larvae of the Globedera rostochiensis pathotype Ro2/3 into pots. In the molecular marker analysis, resistance was investigated with TG689, 57R, Gro1-4 markers. While all cultivars and clones except Bettina were grouped as sensitive in the biotesting study, the H1 resistance gene was detected in Onaran, & Uuml;nlenen, Leventbey, Muratbey, Nahita, Agria, Madeleine, Desiree and Bettina cultivars by molecular marker analysis. H1 and Gro1-4 resistance genes were detected in the PAE 13-08-07, PAE 13-08-08 and PAE 13-08-14 clones used in the experiment. The results showed that clones developed by the Potato Research Institute exhibited highly resistant marker alleles for the Ro2/3 pathotype of G. rostochiensis. The results of phenotyping study and the molecular marker study were not similar.
Distribution of Steinernematids and Heterorhabditids (Rhabditida: Steinernematidae and Heterorhabditidae) in the Southern Anatolia Region of Turkey
(Springer, 2016) Canhilal, Ramazan; Waeyenberge, Lieven; Toktay, Halil; Bozbuğa, Refik; Çetintaş, Ramazan; İmren, Mustafa
This study was conducted to isolate and identify entomopathogenic nematode (EPN) species in different agricultural areas in the Southern Anatolia region of Turkey. Out of 193 soil samples, 20 (10.33%) turned out to be positive for the presence of EPNs, belonging to the genera Steinernema and Heterorhabditis. Morphological and molecular (ITS-RDNA) characterizations were performed for the 20 EPN isolates. Six isolates were identified as H. bacteriophora, 2 as H indica and 6 as S. feltiae. Two isolates were undescribed Steinernemas pecies. H. indica was recorded for the first time in the southern Anatolia region of Turkey. Molecular analysis confirmed the morphological identification of Heterorhabditidae and Steinernematidae species. Comparative analysis of the ITS sequences revealed interspecific polymorphism among the tested isolates. Additionally, intraspecific polymorphism was observed within S. feltiaeand H bacteriophorapopulations, but no intraspecific polymorphism was detected in the H indicapopulation.
Identification and biocontrol potential of entomopathogenic nematodes and their endosymbiotic bacteria in apple orchards against the codling moth, cydia pomonella (l.) (lepidoptera: tortricidae)
(MDPI, 2022) Gümüşsoy, Asım; Yüksel, Ebubekir; Özer, Göksel; İmren, Mustafa; Canhilal, Ramazan; Amer, Mohammed; Dababat, Abdelfattah A.
The codling moth, Cydia pomonella (L.) (Lepidoptera: Tortricidae), is one of the major pests in pome fruit production worldwide. Heavy treatment of the larvae of C. pomonella with insecticides triggered the development of resistance to many groups of insecticides. In addition, the increasing concern about the adverse effects of synthetic insecticides on human health and the environment has led to the development of sustainable and eco-friendly control practices for C. pomonella. The entomopathogenic nematodes (EPNs) (Steinernema and Heterorhabditis spp.) and their endosymbionts (Xenorhabdus and Photorhabdus spp.) represent a newly emerging approach to controlling a wide range of insect pests. In the present study, field surveys were conducted in apple orchards to isolate and identify EPNs and their endosymbionts and evaluate their insecticidal efficacy on the larvae of C. pomonella. EPNs were isolated from 12 of 100 soil samples (12%). Seven samples were identified as Steinernema feltiae (Filipjev, 1934) (Rhabditida: Steinernematidae), whereas five samples were assigned to Heterorhabditis bacteriophora (Poinar, 1976) (Rhabditida: Heterorhabditidae). The pathogenicity of the EPN species/isolates was screened on the last instar larvae of G. mellonella. The two most pathogenic isolates from each EPN species were tested against fifth instar larvae of C. pomonella under controlled conditions. The maximum mortality (100%) was achieved by all EPN species/isolates at a concentration of 100 IJs/larva 96 h after treatment. The endosymbionts of selected H. bacteriophora and S. feltiae species were identified as Photorhabdus luminescens subsp. kayaii and Xenorhabdus bovienii, respectively. The mortality rates ranged between 25 and 62% when the fifth larval instar larvae of C. pomonella were exposed to the treatment of cell-free supernatants of symbiotic bacteria. In essence, the present survey indicated that EPNs and their symbiotic bacteria have good potential for biological control of C. pomonella
Insecticidal effect of entomopathogenic nematodes and the cell-free supernatants from their symbiotic bacteria against different larval instars of Agrotis segetum (Denis & Schiffermuller) (Lepidoptera: Noctuidae)
(Springer, 2022) Yüksel, Ebubekir; İmren, Mustafa; Özdemir, Esengül; Bozbuğa, Refik; Canhilal, Ramazan
Background Entomopathogenic nematodes (EPNs) (Steinernematidae and Heterorhabditidae) and their symbiotic bacteria are pathogenic for a wide range of insect pests and have been used successfully as a biological control agent. Although EPNs are well studied against many agricultural insect pests, the efficacy of their symbiotic bacteria still remains unclear for many insect pests of agricultural importance. In the present study, the virulence of native EPN isolates and their cell-free supernatants of symbiotic bacteria were tested against the 3rd and 4th larval instars of Agrotis segetum (Denis & Schiffermuller) (Lepidoptera: Noctuidae) under laboratory conditions (25 +/- 1 degrees C and R.H. 60%). Results The 4th instar larvae were more susceptible to infective juveniles (IJs) and mortalities over (95%) were achieved by all tested EPN isolates at the concentration of 100 IJs/cm(2) after 72 hrs of exposure. The cell-free supernatants were more effective against the 3rd instar larvae and the highest mortalities were recorded as 42 and 60% in the contact and leaf disc bioassays, respectively. Conclusion The results indicated that the cell-free supernatants can be an ideal application for young larval stages of A. segetum. However, further studies are required to test the effectiveness of both EPNs and the cell-free supernatants of their symbiotic bacteria in field conditions.
Molecular identification and biodiversity of wireworm species, Agriotes spp. Eschscholtz, 1829 (Coleoptera: Elateridae) in major potato cultivated areas of Turkiye
(Entomological Soc Turkey, Ege Univ, 2023) Askar, Arife Gümüş; Yüksel, Ebubekir; Dinçer, Dilek; Özer, Göksel; İmren, Mustafa; Canhilal, Ramazan; Kütük, Halil
Wireworms, Agriotes spp. Eschscholtz, 1829 (Coleoptera: Elateridae) are among the most harmful soil-borne insect pests and significantly reduce potato yields under heavy infestations. The presence of wireworm species on potatoes in potato growing areas is not fully known in the provinces of Turkiye. Therefore, this research aimed to identify wireworms molecularly and evaluate their biodiversity in potato growing ecosystems. Here, the first extensive field survey was carried out in Turkiye's principal potato-growing regions in 2019 and 2020 (Afyon, Bolu, Izmir, Kayseri, Konya, Nigde, and Sivas). Species identification of wireworms was performed using DNA barcoding approach based on the fragment of mitochondrial cytochrome c oxidase subunit I (COI). Samples were collected from 400 potato fields, and 510 larval specimens were obtained. The presence of wireworms was confirmed for Afyon, Bolu, Kayseri, Konya, and Sivas provinces, with an average prevalence of 13.5%, while no positive samples were recovered from Nigde and Izmir. Agriotes sputator (L.,1758) (Coleoptera: Elateridae) was the most prevalent species in surveyed areas. The Shannon index of wireworm species was found to be as 0.59, which implies a low degree of biodiversity of wireworms in potato fields.
Occurrence, identification, and diversity of parasitic nematodes in apple (Malus domestica Borkh.) orchards in the Central Anatolia Region of Turkiye
(Springer Heidelberg, 2023) Yüksel, Ebubekir; İmren, Mustafa; Özer, Göksel; Bozbuğa, Refik; Dababat, Abdelfattah A. A; Canhilal, Ramazan
Root lesion nematodes (RLNs) (Pratylenchus spp.) are one of the most economically important plant parasitic nematodes (PPNs) and are considered one of the leading causative agents of apple diseases that severely decrease yield and quality in apple (Malus domestica Borkh.) orchards. Occurrence, identification, and diversity of PPNs in apple orchards in central Anatolia region of Turkiye have not been well understood. Therefore, in this study, a field survey was conducted in this region to reveal the occurrence, distribution, and diversity of mainly RLNs. The primary analysis of soil and root samples based on morphology revealed that 8 PPN species were present in sampled areas, the most prevalent of which was RLNs. As a result of morphological, morphometric, and molecular studies, a total of 5 species of RLNs [Pratylenchus crenatus Loof, 1960; Pratylenchus neglectus (Rensch, 1924) Filipjev & Schuurmans-Stekhoven, 1941; Pratylenchus penetrans (Cobb, 1917) Filipjev & Schuurmans-Stekhoven, 1941; Pratylenchus thornei Sher & Allen, 1953 and Pratylenchus vulnus Allen & Jensen, 1951] were identified. Phylogenetic analysis with the D2-D3 domain of LSU-rDNA sequences further confirmed the identification. The results indicated that P. thornei was the predominant species in surveyed areas, followed by P. neglectus. The results confirmed the host status of apples for RLNs and demonstrated that RLNs were the most threatening PPNs in the surveyed apple orchards. The necessity of determining the damage caused by plant parasitic nematodes to apple production emerges as an important factor to be done as a future study.
Potential of four Turkish isolates of entomopathogenic nematodes against three major stored products insect pests
(Pergamon-Elsevier Science Ltd, 2019) Yüksel, Ebubekir; Canhilal, Ramazan; İmren, Mustafa
The insecticidal effect of native entomopathogenic nematode (EPN) isolates (Steinernema feltiae UTP-5 isolate, Heterorhabditis bacteriophora UMK-7 isolate, S. feltiae DDKY-11 isolate and H. bacteriophora AVB-15 isolate) recovered from the Cappadocia Region of Turkey were investigated on the adults of S. granarius and R. dominica and the larvae of E. kuehniella at different temperatures (15 degrees C, 20 degrees C and 25 degrees C) and concentrations (250, 500 and 1000 IJs/adult for S. granarius and R. dominica; 10, 50 and 100 IJs/larvae for E. kuehniella) under controlled conditions. Insect mortality was recorded after 4th, 6th and 8th day of exposure time for S. granarius and R. dominica, and 2nd and 4th day for E. kuehniella. The results showed that the efficacy of the isolates at 25 degrees C was generally higher than the ones at 15 degrees C and 20 degrees C. Steinernema feltiae DDKB-17 isolate caused the highest mortality (86%) against S. granarius adults at all temperatures tested. The virulence of the tested isolates on R. dominica adults did not exceed 44% at all temperatures and concentrations tested. UMK-7 and AVB-15 isolates were the most virulent isolates against E. kuehniella larvae with a mortality rate of 100%. Based on the results obtained from the laboratory tests, it appeared that the isolates tested have a good potential for the management of S. granarius, R. dominica and E. kuehniella. (C) 2019 Elsevier Ltd. All rights reserved.
Potential of four Turkish isolates of entomopathogenic nematodes against three major stored products insect pests (vol 83, pg 317, 2019)
(Pergamon-Elsevier Science Ltd, 2019) Yüksel, Ebubekir; Canhilal, Ramazan; İmren, Mustafa
Düzeltme “Potential of four Turkish isolates of entomopathogenic nematodes against three major stored products insect pests”
The Role of Storage Duration and Conditions on the Survival and Pathogenicity of Entomopathogenic Nematodes
(2023) Akı, Onur; Yüksel, Ebubekir; Imren, Mustafa; Bozbuğa, Refik; Canhilal, Ramazan
Entomopathogenic nematodes (EPNs) (Rhabditida: Heterorhabditidae and Steinernematidae) are effective biocontrol agents for many insect pests and are generally stored for a period of time prior to their use in the laboratory or field. However, unfavorable storage conditions have a great impact on the survival and infectivity of EPNs. This study was conducted to determine the optimum storage conditions of infective juveniles (IJs) of four native EPN species (Heterorhabditis bacteriophora FLH-4H, H. indica 216-H, Steinernema feltiae KCS-S, and S. bicornotum MGZ-4S) under laboratory conditions. The survival capability of the IJs was tested at different concentrations (500, 1000, 1500, and 2000 IJs), temperatures (9 and 25 °C) and storage media [double-distilled water (ddH2O), tap water, and sterile Ringer solution]. In general, the survival of IJs of tested EPN species was the highest at the 1st month after treatment (MAT) at the concentrations of 1000 and 1500 IJs and gradually decreased with the increasing storage periods. The survival rates of the IJs of Steirnematids were generally higher than Heterorhabditid species. The highest survival of IJs was generally obtained after 1-month storage in Ringer solution at 9°C while tap water led to poor survival for the IJs at both temperatures tested. The IJs that were stored at 9°C induced higher mortalities on the larvae of Galleria mellonella (L.) (Lepidoptera: Pyralidae). The results showed that the IJs of tested EPN species can remain viable for a longer period of time in Ringer solution at 1000 and 1500 IJs concentrations at 9°C without losing much of their infectivity.
Xenorhabdus and photorhabdus bacteria as potential candidates for the control of culex pipiens l. (diptera: culicidae), the principal vector of west nile virus and lymphatic filariasis
(MDPI, 2023) Yüksel, Ebubekir; Yıldırım, Alparslan; İmren, Mustafa; Canhilal, Ramazan; Dababat, Abdelfattah A.
Vector-borne diseases pose a severe threat to human and animal health. Culex pipiens L. (Diptera: Culicidae) is a widespread mosquito species and serves as a vector for the transmission of infectious diseases such as West Nile disease and Lymphatic Filariasis. Synthetic insecticides have been the prime control method for many years to suppress Cx. pipiens populations. However, recently, the use of insecticides has begun to be questioned due to the detrimental impact on human health and the natural environment. Therefore, many authorities urge the development of eco-friendly control methods that are nontoxic to humans. The bacterial associates [Xenorhabdus and Photorhabdus spp. (Enterobacterales: Morganellaceae)] of entomopathogenic nematodes (EPNs) (Sterinernema spp. and Heterorhabditis spp.) (Rhabditida: Heterorhabditidae and Steinernematidae) are one of the green approaches to combat a variety of insect pests. In the present study, the mosquitocidal activity of the cell-free supernatants and cell suspension (4 x 10(7) cells mL(-1)) of four different symbiotic bacteria (Xenorhabdus nematophila, X. bovienii, X. budapestensis, and P. luminescens subsp. kayaii) was assessed against different development stages of Cx. pipiens (The 1st/2nd and 3rd/4th instar larvae and pupa) under laboratory conditions. The bacterial symbionts were able to kill all the development stages with varying levels of mortality. The 1st/2nd instar larvae exhibited the highest susceptibility to the cell-free supernatants and cell suspensions of symbiotic bacteria and the efficacy of the cell-free supernatants and cell suspensions gradually declined with increasing phases of growth. The highest effectiveness was achieved by the X. bovienii KCS-4S strain inducing 95% mortality to the 1st/2nd instar larvae. The results indicate that tested bacterial symbionts have great potential as an eco-friendly alternative to insecticides.

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