Selcuk Hazir

Diversity and distribution of entomopathogenic nematodes (Steinernematidae, Heterorhabditidae) in South Africa

A total of 1506 soil samples from different habitats in seven geographic regions of South Africa were evaluated for the presence of entomopathogenic nematodes (EPN). Nematodes were isolated from 5% of the samples. Among the steinernematids, four Steinernema sp. were recovered including Steinernema khoisanae and three new undescribed species. Although steinernematids were recovered from both humid subtropical and semiarid regions, this family accounted for 80% of EPN recovered from the semiarid climate zones characterised by sandy, acidic soils. Eight isolates of S. khoisanae were recovered from the Western Cape province. One of the new undescribed steinernematids (Steinernema sp. 1) was recovered only from the Free State and KwaZulu-Natal provinces where humid subtropical conditions prevail and soils are generally less acidic with higher clay content. A high level of adaptation, however, was noted with Steinernema sp. 2, which was recovered from a wide range of soil conditions and habitats ranging from semiarid (Western Cape province) to humid subtropical (KwaZulu-Natal province). A third undescribed steinernematid, Steinernema sp. 3, seemed better adapted to heavier soils with more than 80% of isolates recovered from fruit orchards in the Free State province. Heterorhabditis bacteriophora was the only heterorhabditid recovered during this survey. This species was particularly prevalent in four provinces ranging from humid subtropical to semiarid regions. Isolation of EPN directly from insect cadavers included Steinernema sp. 2 and one H. bacteriophora from an unidentified white grub (Scarabaeidae) cadaver (i.e., dual infection) and H. bacteriophora from the black vine weevil, Otiorhynchus sulcatus.

Diversity and distribution of entomopathogenic nematodes (Rhabditida: Steinernematidae and Heterorhabditidae) in Turkey

The diversity and distribution of entomopathogenic nematodes in thefamilies Steinernematidae and Heterorhabditidae were assessed throughout anextensive soil survey in Turkey during 1999 and 2000. Entomopathogenic nematodeswere recovered from six out of seven regions sampled, with 22 positive sites(2%) out of 1080 sites sampled. A single nematode isolate was recovered at eachof the positive sites, of which 15 were steinernematid isolates and seven wereheterorhabditid isolates representing a total of four species. Based onmorphometric and molecular data, the nematode species were identified asHeterorhabditis bacteriophora, Steinernemafeltiae, S. affine, andSteinernema n. sp. The most common species was S.feltiae, which was isolated from 10 sites in six regions, followed byH. bacteriophora from seven sites in five regions,S. affine from four sites in two regions, andSteinernema n. sp. from one site. Heterorhabditisbacteriophora and S. feltiae have been found inmany parts of the world, whereas S. affine, so far, hasonly been recovered in Europe until our survey. Steinernemaaffine was isolated from the European (Marmara) as well as theAsiatic region (Middle Anatolia) of Turkey. A new undescribedSteinernema sp. was isolated from the most eastern region(East Anatolia) of Turkey. Soils of the positive sites were classified as sandy,sandy loam, or loam (68.2%) and sandy–clay–loam or clay loam (31.8%) and the pHranged from 5.6 to 7.9. The habitats from which the entomopathogenic nematodeswere isolated were broadly classified as disturbed (59.1%), which includedagricultural fields and poplar planted for lumber and wind breaks, andundisturbed (40.9%), which included pine forest, grassland, marsh and reed sites.Steinernema feltiae, S. affine, andH. bacteriophora were recovered from both disturbed andundisturbed habitats. The new Steinernema sp. was recoveredfrom grassland. Our survey showed that these nematodes occur widely throughoutTurkey, but at a frequency below that reported for other parts of the world.

Two new subspecies of Photorhabdus luminescens, isolated from Heterorhabditis bacteriophora (Nematoda: Heterorhabditidae): Photorhabdus luminescens subsp. kayaii subsp. nov. and Photorhabdus luminescens subsp. thracensis subsp. nov.

Bacterial isolates from nematodes from Turkish soil samples were initially characterized by molecular methods and seven members of the genus Photorhabdus identified to the species level, using riboprint analyses and metabolic properties. Strain 07-5 (DSM 15195) was highly related to the type strain of Photorhabdus luminescens subsp. laumondii DSM 15139T, and was regarded a strain of this subspecies. Strains 1121T (DSM 15194T), 68-3 (DSM 15198) and 47-10 (DSM 15197) formed one, strain 39-8T (DSM 15199T), 39-7 (DSM 15196) and 01-12 (DSM 15193) formed a second cluster that branched intermediate the three subspecies of Photorhabdus luminescens. Based upon moderate 16S rRNA gene sequence similarities and differences in metabolic properties among themselves and with type strains of the three subspecies we consider the two clusters to represent two new subspecies of Photorhabdus luminescens for which the names Photorhabdus luminescens subsp. kayaii, type strain 1121T (DSM 15194T, NCIMB 13951T), and Photorhabdus luminescens subsp. thracensis subsp. nov., type strain 39-8T (DSM 15199T, NCIMB 13952T) are proposed.

Antibacterial Activity and Chemical Composition of Turkish Propolis

Abstract The antibacterial activities of propolis samples have been examined in vitro, according to the principles accepted for the determination of a similar activity of antibiotics with the use of solid and liquid media. It has been found that propolis extracts showed antibacterial activity through a range of commonly encountered gram positive cocci (5. aureus, beta hem. Streptococus), but had weak activity against gram negative bacteria (E . coli, P. aeruginosa). GC/MS analysis showed that propolis samples contain a variety of chemical compounds in cluding aromatic compounds, fatty acid esters and sesquiterpenes.

Natural enemies of natural enemies: the potential top‐down impact of predators on entomopathogenic nematode populations

Top‐down population regulation can influence the success of biological control agents when they are released into the field. Entomopathogenic nematodes (EPNs) are used commonly in biological control programmes, but their efficacy suffers from poor persistence. Although abiotic soil conditions have been shown to reduce EPN persistence, consumption of infected insects by scavengers and of infective juvenile (IJ) nematodes by predators may also regulate these populations. In the present study, the effects of different soil arthropods on EPNs in laboratory conditions were measured. It was hypothesised that arthropods commonly found in soil communities where EPNs are applied would consume cadavers of insects parasitised by the nematodes and the IJs themselves. Some species of scavengers consume EPN‐infected insects. Crickets (Gryllus bimaculatus De Geer), American cockroaches [Periplaneta Americana (Linnaeus)], ants [Tetramorium chefketi Forel and Pheidole pallidula (Nylander)], earwigs (Labidura riparia Pallas), mites (Sancassania polyphyllae Zachvatkin), and springtails (Sinella curviseta Brook and Folsomia candida Willem) have different responses to nematode‐killed insects. Results suggested that ants (T. chefketi), cockroaches, mites, and earwigs fed on Steinernema‐killed insects whereas neither crickets nor springtails consumed them. In the second part of the study, experiments were conducted to determine whether mites and springtails consumed IJ EPNs. Results showed that S. polyphyllae mites do not consume infective juveniles in soil, whereas both springtail species consumed significant numbers of the IJs. Top‐down regulatory processes can be a limiting factor for EPN populations under laboratory conditions. Both host cadavers and IJs are consumed (albeit by different arthropods), so these results may help explain the difficulties associated with the persistence of EPN application to soil.

Food preference of Sancassania polyphyllae (Acari: Acaridae): living entomopathogenic nematodes or insect tissues?

Abstract Sancassania polyphyllae (Acari: Acaridae) adult female mites will feed on insect cadavers and infective juveniles (IJs) of entomopathogenic nematodes. Our objective was to determine whether S. polyphyllae has a food preference when offered a choice between tissues of Polyphylla fullo (Coleoptera: Scarabaeidae) or Galleria mellonella (Lepidoptera: Pyralidae), or IJs of Steinernema feltiae (Rhabditida: Steinernematidae) or IJs of Heterorhabditis bacteriophora (Rhabditida: Heterorhabditidae). When offered a choice between no food and one of the different food sources, P. fullo, G. mellonella or S. feltiae IJs, S. polyphyllae had a significant preference for food sources compared to no food. When it was offered either no food or H. bacteriophora, there was no significant difference in the mite distribution. When offered two different food choices, P. fullo or G. mellonella, P. fullo or S. feltiae, and P. fullo or H. bacteriophora, the mite showed significant preferences for P. fullo larvae. In S. feltiae vs. G. mellonella and S. feltiae vs. H. bacteriophora experiments, S. polyphyllae showed significant a preference for S. feltiae. In three-choice experiments, S. polyphyllae had a preference for P. fullo, followed by S. feltiae, G. mellonella and H. bacteriophora, respectively. Our data confirm, in part, our hypothesis that when offered different food choices, this mite species prefers tissues of its phoretic host, P. fullo over lepidopteran host tissues or living IJs. Based on these laboratory data, H. bacteriophora should be used as a biological control agent against P. fullo over a Steinernema species.

Trophic Relationships of Entomopathogenic Nematodes in Agricultural Habitats

Entomopathogenic nematodes (EPNs) play several roles in the soil ecosystem. While EPNs are generally thought of in the context of reducing the density of pest populations when they are applied, they are also natural components of soil food webs and exert considerable influence on the population dynamics of many players in the system in addition to the intended targets of biological control efforts (Hodson, Siegel, & Lewis, 2012). They are lethal parasites of insects, but not all of the species they infect are the targets for which they are applied. They are also prey and hosts to a variety of other soil organisms. Here, we attempt explain the fate of EPNs after they are applied to soil in the context of the complicated interactions among members of soil food webs.

Diversity and distribution of nematodes associated with wild bees in Turkey.

The diversity and distribution of nematodes associated with soil-dwelling bees in Turkey were examined. A total of 9456 wild bees were collected and dissected to detect nematodes and 173 female and 22 male bees were positive for an overall association level of 2.1%. Halictus resurgens was the most commonly collected bee found with nematodes with 82 and 19 nematode-infested female and male bees, respectively. Sixty-four of the nematode isolates were recovered from the Black Sea region (32.8%), 59 from Central Anatolia (30.3%), 35 from the Mediterranean (17.9%), 24 from the Aegean (12.3%), six from Eastern Anatolia (3.1%) and seven from Southeast Anatolia (3.6%). In 3.9% of all bees of the family Halictidae (173/4417), nematodes were observed in the Dufours gland, poison sac, oviduct, bursa copulatrix and abdominal cavity of dissected female bees and from the genital capsule of male bees. Only abdominal glands of 0.6% (21/3279) of female bees from the family Andrenidae were observed with nematodes. Nematodes were isolated from the genital capsule of one male bee from the family Melittidae. The poison sac was the organ where nematodes were most often observed but the highest number of nematodes occurred in the abdominal cavity. Most nematodes were in the dauer juvenile stage making identification difficult. Sequence analysis using LSU D2/D3 domains and a fragment of mitochondrial cytochrome oxidase I (mtCOI) allowed for putative identification of nematode isolates. Two or three species of Bursaphelenchus ( B. anatolius , B. debrae , and potentially one or more cryptic species closest in morphology to B. anatolius ) were isolated from six different halictid species (two genera; Halictus and Lasioglossum ) from different geographical regions in Turkey. An additional four species of diplogastrid were recovered from the abdominal glands of andrenids; a new species of Koerneria from Andrena limata and a separate sister clade to Mononchoides composed of three other possible species from four Andrena species. An additional unknown species in the order Tylenchida (near Allantonematinae) was recovered from three species of Lasioglossum . No molecular data were obtained from three mermithid juveniles from two Andrena and one Halictus species because samples were lost in transit. The associations of most nematode isolates with bees of the families Halictidae and Andrenidae were hypothesised to be phoretic, whereas the tylenchid and mermithids were parasites.

Laboratory evaluation of Turkish entomopathogenic nematodes for suppression of the chestnut pests, Curculio elephas (Coleoptera: Curculionidae) and Cydia splendana (Lepidoptera: Tortricidae).

Abstract The lepidopteran, Cydia splendana, and the coleopteran, Curculio elephas, are the most serious pests of chestnut fruit in Turkey. We evaluated the biological control potential of three Turkish entomopathogenic nematode species, Steinernema feltiae, S. weiseri and Heterorhabditis bacteriophora, against the last instar larvae of C. splendana and C. elephas, both of which occur in the soil from fall (October–November) until mid-summer (August). The optimal temperature for infection, time to death of the hosts, and reproductive potential of the nematodes were determined at 10, 15, 20 and 25°C for both pest species. Cydia splendana was more susceptible to nematode infection than C. elephas. Temperature had a significant effect on the infectivity and development of entomopathogenic nematodes. The cold-adapted S. weiseri and S. feltiae were the most virulent species at 10 and 15°C, whereas the warm-adapted H. bacteriophora was the most effective at 20 and 25°C. In soil pot experiments conducted at 15°C, S. weiseri was the most virulent species against C. elephas and C. splendana. However, our data show that C. elephas larvae had a lower and C. splendana larvae had a higher susceptibility to the nematode species tested. Accordingly, we recommend that future efforts of using entomopathogenic nematodes, especially S. weiseri, be directed against C. splendana and that there be a continued effort to find more virulent nematode isolates against larvae of C. elephas.

Isolation and identification of entomopathogenic nematodes (Nematoda: Rhabditida) from the Eastern Black Sea region and their biocontrol potential against Melolontha melolontha (Coleoptera: Scarabaeidae) larvae

A survey was conducted for entomopathogenic nematodes in various agricultural fields in the Eastern Black Sea region of Turkey. A total of 77 soil samples were collected from 15 distinct geographic areas during 2010-2011. Seven entomopathogenic nematode isolates (ZET02, ZET04, ZET09, ZET28, ZET31, ZET35, and ZET76) were detected from the soil samples (9.1% positive) using the Galleria baiting technique. Morphological and molecular characterizations of the isolates were performed for species identification. Five isolates were identified as Heterorhabditis bacteriophora (ZET02, ZET04, ZET09, ZET28, and ZET35) and 2 isolates were identified as Steinernema feltiae (ZET31 and ZET76). The efficacy of all isolates was tested on Melolontha melolontha larvae in plastic boxes and pot experiments. Different concentrations of nematodes at 0, 500, 1000, or 2000 infective juveniles (IJs)/mL and 2 different temperature regimes (15 and 25 °C) were used. One hundred percent mortality was obtained from the ZET09 and ZET35 isolates at a concentration of 2000 IJs/mL at 25 °C. The same isolates also provided 100% protection with 100 IJs/cm2 in strawberry planted pot experiments.