Zygomycota: Entomophthorales) Species Infecting Dermaptera and Psocodea

Available online at www.sciencedirect.com Journal of INVERTEBRATE PATHOLOGY Journal of Invertebrate Pathology 97 (2008) 193–196 www.elsevier.com/locate/yjipa Short Communication First and southernmost records of Hirsutella (Ascomycota: Hypocreales) and Pandora (Zygomycota: Entomophthorales) species infecting Dermaptera and Psocodea

A.V. Toledo a,*, R.A. Humber b, C.C. Lo´pez Lastra a

a Centro de Estudios Parasitolo´gicos y de Vectores (CEPAVE) UNLP-CONICET, Calle 2 Nro. 584 (1900), La Plata, Buenos Aires, Argentina b USDA-ARS Plant Soil and Nutrition Laboratory, Tower Road, Ithaca, NY 14853, USA

Received 4 May 2007; accepted 5 September 2007 Available online 14 September 2007

Abstract

We present the first and southernmost records of the fungi Hirsutella strigosa Petch, H. citriformis Speare (Ascomycota: Hypocreales), and Pandora nouryi (Remaudie`re & Hennebert) Humber (Zygomycota: Entomophthorales) infecting Doru lineare (Eschscholtz) (Der- maptera: Forficulidae), Ectopsocus californicus (Banks) (Psocodea: Ectopsocidae), and Heterocaecilius sp. (Psocodea: Pseudocaeciliidae), respectively. This is the first report of naturally occurring pathogenic fungi infecting Psocoptera, and it is also the first report of P. nouryi from any host outside of the Aphididae. The three fungal species were morphologically described from their host insects and from microscopic preparations. Attempts to obtain pure fungal isolates were unsuccessful but slides and photographs of these fungi were preserved and deposited in mycological collections as herbarium material. 2007 Elsevier Inc. All rights reserved.

Keywords: Bark lice; Booklice; Dermaptera; Earwigs; Entomopathogenic fungi; Entomophthorales; Hypocreales; Psocids; Psocodea; South America

1. Introduction A few species of entomopathogenic fungi have been recorded from Dermaptera worldwide. Among these fungi Some earwigs (Insecta: Dermaptera) feed on living are Beauveria bassiana (Balsamo-Crivelli) Vuillemin (Asco- plants and often become pests in greenhouses and field mycota: Hypocreales) found on Forficula auricularia L., F. crops (Fulton, 1924; Crumb et al., 1941; Behura, 1956; africana Dohrn, and Doru lineare (Eschscholtz) (Forficuli- Lamb, 1974, 1976). The family Forficulidae comprises 10 dae) (http://arsef.fpsnl.cornell.edu; Mariani et al., 1996; subfamilies among which the Forficulinae is one of the Toledo et al., 2007), Metarhizium anisopliae (Metschnicoff) most important. This subfamily includes three genera rep- Sorokin (Ascomycota: Hypocreales) infects D. lineare resented in the Neotropical region: Forficula (Linn.), Skal- (Mariani et al., 1996), and Zoophthora forficulae (Giard) istes Burr, and Doru Burr; the latter two genera are A. Batko (Zygomycota: Entomophthorales), which infects endemic to the Neotropics. Some booklice (Insecta: Psoco- F. auricularia (Pell et al., 2001). Nevertheless, no records of dea) have an affinity for grain products (Turner, 1987) but entomopathogenic fungi affecting Psocodea are known at do not cause direct damage to stored products even though the present time. populations commonly build up to unacceptable levels The objectives of our study were to report the presence (Turner, 1994). of pathogenic fungi naturally infecting insects in the orders Dermaptera and Psocodea from Argentina, and to provide * Corresponding author. Fax: +54 221 423 2327. morphological characterizations of the fungal species E-mail address: [email protected] (A.V. Toledo). found.

2. Materials and methods (1.4 ± 1.1 lm; n =7)(Fig. 1b and c). Conidia hyaline, aseptate, smooth, with the shape of a citrus segment, 7.4– Adults of D. lineare were collected in December 2004 on 11.2 lm (8.4 ± 0.2 lm) · 4.9–6.2 lm (5 ± 0.05 lm; Zea mays L. (Fam. Poaceae) plants at Arana, Buenos Aires n = 23) (Fig. 1c), occasionally arising in small groups (2 province (35 00 19.800 S–57 550 3000 W), while Heterocaeci- or 3) embedded in a pigmented (brownish) mucous sheath lius sp. (Psocodea: Pseudocaeciliidae) and Ectopsocus cali- at the apex of each neck. fornicus (Banks) (Psocodea: Ectopsocidae) were found in Specimens examined: Adult of earwig D. lineare col- August 2005 and November 2006, respectively, both on lected on Zea mays, Arana, Buenos Aires, Argentina, the underside of leaves of Quercus ilex L. (Fam. Fagaceae) December 15, 2004. Herbarium material access number at La Plata, Buenos Aires Province (34 550 4900 S–57 560 LPSC 47783. 3200 W). A total of three dead insects (one of each species) with external signs of mycosis was collected from their host 3.2. H. citriformis speare (Ascomycota: Hypocreales) plants, placed into sterilized plastic containers, and then processed in the laboratory. Healthy insects were also col- Mycelia brownish more or less covering the host. Synne- lected and fixed in 70% ethanol for their later taxonomic mata, slender, and simple, composed of a compact bundle identification. Measurements of fungal structures (myce- of longitudinal hyphae, pubescent from the conidiogenous lium, conidiogenous cells, and conidia) from host insects cells (Fig. 1d). Conidiogenous cells forming a moderately were made to enable specific identifications. Microscopic compact layer over the surface of the synnema, mostly aris- characters were described from material mounted in lac- ing as lateral cells produced over the whole length of the tophenol/cotton blue (0.01% w/v) or lactophenol/aceto- synnema and are intercalary along the length of the myce- orcein (1% w/v), and observed with phase contrast optics lial strands (Fig. 1e and f), monophialidic, with ellipsoid on an Olympus CH3 microscope. Fungal preparations base tapering abruptly to long, phialidic sterigmata, 20.8– were photographed using an Olympus CH3 microscope fit- 32.7 lm long (28.3 ± 0.9 lm; n = 21), 3.5–5.9 lm wide at ted with an Olympus SC35 camera; infected insect hosts the base (4.6 ± 0.1 lm; n = 21) (Fig. 1e and f). Sterigmata were photographed using an Olympus SZ-PT stereo micro- about 10.9–24.8 lm long (19.2 ± 1.2 lm; n = 20). Conidia scope fitted with a Sony CyberShot camera. Semi-perma- hyaline, aseptate, smooth-walled, fusiform or elliptical, nent slides were made according to Humber (1997) and 5.9–7.9 lm (6.5 ± 0.1 lm) · 1.9–2.9 lm (2.5 ± 0.1 lm; deposited as herbarium material in the Mycological Collec- n = 26) (Fig. 1f). Conidia are solitary or occasionally tion of the Institute of Botany Carlos Spegazzini (LPSC, paired. La Plata, Buenos Aires, Argentina). Photographs of the Specimens examined: Adult of psocid E. californicus col- slides were also deposited as herbarium material in the lected from the underside of living leaf of a Q. ilex tree, La LPSC. Plata, Buenos Aires, Argentina, November 2, 2006. Her- barium material access number LPSC 47785. 3. Results 3.3. Pandora nouryi (Remaudie`re & Hennebert) Humber Three fungal species were identified from infected (Zygomycota: Entomophthorales) insects. Attempts to obtain pure cultures on Sabouraud dextrose agar + 1% yeast extract (SDA-Y 1%) with Mycelia yellowish, covering the entire insect abdomen, 40,000 units/ml penicillin G (Merck, Germany) and with presence of cylindrical cystidia (Fig. 1g). Cadaver 80,000 units/ml streptomycin (Parafarm, Argentina) were attached to the substrate by a few single rhizoids with dis- not successful. Nevertheless, measurements and fungal coid terminal holdfasts emerging from the host thorax descriptions were made from material mounted in lactoph- (Fig. 1h). Conidiophores branched 4.9–11.2 lm diam. enol/cotton blue (0.01% w/v) or lactophenol/aceto-orcein (7.4 ± 0.3 lm; n = 24). Primary conidia short-ovoid, bituni- (1% w/v) on microscope slides. cate, uninucleate, without over papillar neck and with slightly convex, rather broad papilla, 13.6–19.8 lm 3.1. Hirsutella strigosa petch (Ascomycota: Hypocreales) (16 ± 0.2 lm) · 7.4–13.6 lm (10.3 ± 0.2 lm; n = 40) (Fig. 1i and j). Resting spores not observed. Mycelia tawny brown, septate, branched and anastomo- Specimens examined: Adult of psocid Heterocaecilius sp. sing, emerging from host body through intersegmental collected from the underside of living leaf of a Q. ilex tree, membranes and growing close to the insect body La Plata, Buenos Aires, Argentina, August 28, 2005. Her- (Fig. 1a), with hyphae of 2.5–3.7 lm diam. (2.5 ± 1.1 lm; barium material access number LPSC 47784. n = 7). Conidiophores reduced to sessile conidiogenous cells, or occasionally conidiogenous cells borne on a single 4. Discussion stalk cell, arising singly. Conidiogenous cells monophialidic, 24.8–42.8 lm long (35 ± 5.9 lm; n = 7), subcylindrical, In addition to it having distinctively pigmented (brown) 3.7–4.9 lm diam. at the base (4.7 ± 1.1 lm; n = 7), taper- vegetative hyphae, H. strigosa clearly differs from H. guy- ing gradually to 1.2–2.5 lm wide with a long straight neck ana Minter and Brady (which produces a distinctive erect A.V. Toledo et al. / Journal of Invertebrate Pathology 97 (2008) 193–196 195

Fig. 1. (a) Adult of Doru lineare infected with Hirsutella strigosa. (b) Conidiogenous cells of H. strigosa. (c) Conidiogenous cells and conidia of H. strigosa. (d) Adult of Ectopsocus californicus infected with Hirsutella citriformis. Note the slender synnemata. (e–f) Conidiogenous cells and conidia of H. citriformis arising from synnemata. (g–h) Adult of Heterocaecilius sp. infected with Pandora nouryi. Note the presence of cystidia and rhizoids (arrows). (i) Bitunicate primary conidium of P. nouryi. (j) Primary conidia of P. nouryi. Scale bar: (a): 0.2 cm, (b): 17.5 lm, (c): 17 lm, (d): 0.7 mm, (e): 13 lm, (f): 13 lm, (g): 0.3 mm, (h): 0.3 mm, (i): 8 lm, (j): 9 lm.

conidiophore bearing whorls or clusters of conidiogenous the Argentinean fungus had hyphae with smaller diameters cells) and H. verticillioides Charles (whose conidiogenous than those described by these authors (up to 5-lm wide). cells are produced frequently in verticils), from H. besseyi The structure of the synnemata, conidiogenous cells, Fisher (which is frequently polyphialidic), and from H. and conidia of H. citriformis formed on the mycotized pso- illustris Minter and Brady (whose conidia are longer, 50– cid was similar to the characteristics of this fungal species 100 lm long) (Minter and Brady, 1980). Our observations as originally described from both Leptopharsa constricta of morphological features of H. strigosa agree with those (Champion) and Corythuca ulmi Osborn & Drake (Hemip- previously reported by Minter and Brady (1980) although tera: Tingidae) (Mains, 1951). The necks of the conidioge- 196 A.V. Toledo et al. / Journal of Invertebrate Pathology 97 (2008) 193–196 nous cells of the Argentinean fungus were shorter than the sion, Natural Science Museum of La Plata, Buenos Aires, 30–40 lm length described by Mains (1951). Argentina) for earwigs identification, and Dr. V. Rosato The shape and measurements of Pandora nouryi primary (Institute of Botany Carlos Spegazzini, La Plata, Buenos conidia forcibly discharged from infected psocid were coin- Aires, Argentina) for the herbarium material preservation. cident with those described by Bałazy (1993), (12) 15– This study was partially supported by the National Re- 16.5 (20) · (7) 8–10.5 (12) lm. P. nouryi shares some search Council of Argentina (CONICET). similar characteristics with P. neoaphidis (Remaudie`re & Hennebert) Humber and P. kondoiensis (Milner) Humber: References For example, conidiophores are digitately branched, cystidia conical or cylindrical, and rhizoids monohyphal with Bałazy, S., 1993. Flora of Poland (Flora Polska), Fungi (Mycota). Polish. distinct disc-like expansion at end, although the three fun- Acad. Sci., W. Szafer Inst. Botany, Krako´w. 24, 1–356. Behura, B.K., 1956. The biology of the European earwig Forficula gal species can be differentiated by the shapes and sizes of auricularia. L. Ann. Zoo. 1, 119–142. its primary conidia. P. nouryi primary conidia are short- Cameron, S.L., Beckenbach, A.T., Dowton, M., Whiting, M.F., 2006. ovoid, (12) 15–16.6 (20) · (7) 8–10.5 (12) lm, with Evidence from mitochondrial genomics on interordinal relationships in no obvious neck-like extension of the spore above the insects. Arthropod Systematics and Phylogeny 64, 27–34. slightly convex rather broad basal papilla. P. neoaphidis Crumb, S.E., Eide, P., Bonn, A., 1941. The European earwig. Tech. Bull. U.S. Dep. Agric., 776. presents ovoid, ellipsoidal to subcylindrical primary coni- Fulton, B.B., 1924. Some habits of earwig. Ann. Ent. Soc. 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Parental behavior in the Dermaptera with special reference to Forficula auricularia (Dermaptera, Forficulidae). Can. Ent. to mention that Psocoptera and Phthiraptera (biting and 108, 609–619. sucking lice) (currently classified as Psocodea) along with Lord, J.C., Howard, R.W., 2004. A proposed role for the cuticular fatty Thysanoptera (thrips) and Hemiptera are closely related amides of Liposcelis bostrychophila (Psocoptera: Liposcelidae) in to hemipteroid insects. The synapomorphies that unite preventing adhesion of entomopathogenic fungi with dry-conidia. these four orders are the absence of cerci, four Malpighian Mycopathologia 158, 211–217. Mains, E.B., 1951. Entomogenous species of Hirsutella, Tilachlidium and tubules, absence of sternum I, one concentrated abdominal Synnematium. Mycologia 43, 691–718. ganglion, lacinial stylets, and spermatozoa with two fla- Mariani, R., Vera, L., Virla, E., 1996. Aportes al conocimiento de Doru gella (Wheeler et al., 2001; Cameron et al., 2006). With this lineare (Eschs., 1822) (Dermaptera, Forficulidae), un insecto de research we add a new order of insects into the natural host importancia agrono´mica en el Noroeste Argentino. CIRPON, Rev range list of this entomophthoralean fungus species. On the Invest. 4, 13–18. Minter, D.W., Brady, B.L., 1980. Mononematous species of Hirsutella. other hand, Lord and Howard (2004) have observed that Trans. Br. Mycol. Soc. 74, 271–282. Liposcelis bostrychophila (Psocoptera: Liposcelidae) has a Pell, J.K., Eilenberg, J., Hajek, A.E., Steinkraus, D.C., 2001. Biology, high tolerance for entomopathogenic fungi. The authors ecology and pest management potential of Entomophthorales. In: speculate that these insects may have evolved secretions Butt, T.M., Jackson, C., Magan, N. (Eds.), Fungal biological control of amphiphilic cuticular compounds as a heretofore-unre- agents: progress, problems and potential. CABI Publ., Oxon, pp. 306– 389. ported antifungal defense as an adaptation to its fungus- Toledo, A.V., De Remes Lenicov, A.M.M., Lo´pez Lastra, C.C., 2007. rich habitats. Perhaps this could be one of the reasons by Pathogenicity of fungal isolates (Ascomycota: Hypocreales) against which there are no records of fungal infections in this order Peregrinus maidis, Delphacodes kuscheli (Hemiptera: Delphacidae) and of insects. Dalbulus maidis (Hemiptera: Cicadellidae), vectors of corn diseases. Mycopathologia 163, 225–232. Turner, B.D., 1987. Forming a clearer view of L. bostrychophilus. Environ Acknowledgments Health. 95, 9–13. Turner, B.D., 1994. Liposcelis bostrychophila (Psocoptera: Liposcelidae), a We wish to thank Dr. A. Garcıá Aldrete (Biology Insti- stored food pest in the UK. Int. J. Pest Managem. 40, 179–190. tute, UNAM. Zoology Department, D.F., Me´xico) for Wheeler, W.C., Whiting, M., Wheeler, Q.D., Carpenter, J.M., 2001. The psocids identification, Dr. R. Mariani (Entomology Divi- phylogeny of the extant Hexapod orders. Cladistics 17, 113–169.