Trichomycetes from Newfoundland, including Gros Morne

Trichomycetes from Newfoundland, includingGros Morne National Park

D.B. Strongman

Abstract: Twenty-one taxa of trichomycetes are recorded from aquatic insects collected from sites on the east and westcoasts of Newfoundland. Three new species, Smittium colboi Strongman sp. nov., Smittium georgense Strongman sp. nov.,and Stachylina forantipes Strongman sp. nov., are described. Spartiella barbata Tuzet & Manier ex Manier is reported forthe first time in North America, and six species are new geographic records for Newfoundland. Of these six, three are newrecords for Atlantic Canada. Pennella hovassi Manier ex Manier was recovered from sites in eastern Newfoundland, andtaxonomically important features of this and other species in the genus are discussed.

Key words: aquatic insects, gut fungi, Harpellales, Kickxellomycotina, new species, Zygomycota.

Resume : L’auteur rapporte vingt et un taxons de trichomycetes provenant d’insectes aquatiques recoltes sur des sites lo-calises sur les cotes est et ouest de Terre-Neuve. Il decrit trois nouvelles especes, Smittium colboi Strongman sp. nov.,Smittium georgense Strongman sp. nov. et Stachylina forantipes Strongman sp. nov.. Il rapporte le Spartiella barbatapour la premiere fois en Amerique du Nord, et six especes representent de nouvelles mentions geographiques pour Terre-Neuve. Parmi ces six entites, trois constituent de nouvelles mentions pour le Canada Atlantique. L’auteur a recolte le Pen-nella hovassi Manier ex Manier sur des sites de l’est de Terre-Neuve et il discute de caracteristiques taxonomiques impor-tantes chez cette espece, ainsi que d’autres especes de ce genre.

Mots-cles : insectes aquatiques, champignons intestinaux, Harpellales, Kickxellomycotina, nouvelles especes, Zygomycota.

[Traduit par la Redaction]

Introduction

The geology of Newfoundland (NL) is diverse. Over 500million years ago, the eastern and western regions of the is-land were separated by an ancient ocean (Iapetus) and asthis ocean closed, the seabed was pushed up over the Cana-dian shield, amalgamating the terranes once separated by Ia-petus into the island of Newfoundland (Owen 1991). Thisgeological origin suggests that the western coast has NorthAmerican affinities, the central part of the island originatesfrom oceanic crustal rock associated with Iapetus, and theeastern region shares its geological origin with what is nowEurope. This unique geological foundation of the westerncoast of NL was the basis for the designation of Gros MorneNational Park as a UNESCO World Heritage Site. Theaquatic systems in NL are influenced by this varied geolog-ical environment. For example, stream pH in the Tablelandsarea with high limestone content is 8 or higher, whereas thewaters flowing through the granitic mountains in the north-ern part of Gros Morne National Park and the watershedsaround St. John’s, in the east, have a much lower pH (e.g.,5–6).

A detailed overview of aquatic insect diversity in NL wasgiven by Larson and Colbo (1983). They describe theaquatic insect diversity for some groups, such as mayflies,

as impoverished relative to the adjacent mainland areas ofLabrador, Quebec, and Nova Scotia, but note that there aremany gaps in the information about insect diversity on theisland. Larson and Colbo (1983) suggested that the mainbarriers to insect colonization were habitat characteristicson the island, rather than physical barriers between NL andthe nearby mainland (e.g., Labrador) sources of aquatic in-sects. Adler et al. (2005) also addressed colonization of is-lands by dipteran insects and concluded that open waters ofmore than 100 km would likely reduce colonization by fly-ing insects originating on the mainland. At its northern tip,NL is much closer to the mainland than this, with the Straitof Belle Isle separating the island from Labrador to thenorth by as little as 15 km. Larson and Colbo (1983) notethat there are few insect species found only in NL, and thespecies with low vagility (e.g., mayflies and stoneflies) thatexist there show, generally, an east–west axis of distributionacross the continent, while more vagile species, such as dip-terans, have a more north–south distribution pattern. Apartfrom some local differences for a few species, the distribu-tion pattern of aquatic insects of NL is fairly uniform, withmost species widely dispersed on the island (Larson andColbo 1983).

There are two reports of symbiotic microorganisms (tri-chomycetes) from the guts of insects collected in NL (Frost

Received 13 August 2010. Accepted 8 October 2010. Published on the NRC Research Press Web site at botany.nrc.ca on 20 December2010.

D.B. Strongman. Department of Biology, Saint Mary’s University, 923 Robie Street, Halifax, NS B3H 3C3, Canada (e-mail:[email protected]).

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Botany 88: 1011–1022 (2010) doi:10.1139/B10-073 Published by NRC Research Press

and Manier 1971; Lichtwardt et al. 2001b). These collec-tions were made mostly near St. John’s on the eastern sideof the island, with eight species of trichomycetes reported.Little is known about the occurrence of these endosymbiontsin most other parts of NL, although the aquatic habitats andinsect diversity there are rich and varied (South 1983).

This paper adds to the diversity information on trichomy-cetes in NL by describing three new species, Smittium col-boi Strongman sp. nov. and Smittium georgense Strongmansp. nov. from collections near St. John’s, and Stachylina for-antipes Strongman sp. nov. from a site in Gros Morne Na-tional Park on the western coast of NL. Twenty-one taxaare listed, including a new continental record (Spartiellabarbata Tuzet & Manier ex Manier). Easterly geographicdistribution range extensions for six other trichomycetes arereported.

Materials and methodsFour lotic systems (streams) were sampled near St. John’s

on 9 November 2006 (Table 1, NL1–4) and additional sam-ples from two of these sites (NL1, NL2) were collected byMurray Colbo (Department of Biology, Memorial Universityof Newfoundland, St. John’s, NL) on 3 June 2007 (Table 1)and sent to my laboratory at Saint Mary’s University.Aquatic insects were also collected from six streams withinGros Morne National Park on 10, 12, and 13 September2008 (Table 1). The site characteristics, physical data re-corded, and coordinates for all sites are shown in Table 1.

Immature stages of aquatic insects were collected withaquatic dip nets (30 cm gape, 0.5 mm mesh) by disturbingthe substrate by foot (kick-sampling) and capturing the dis-lodged hosts in the net. Specimens were also hand-pickedfrom rocks and plant material removed from aquatic habi-tats. The guts of the insect specimens were dissected in

drops of water in glass Petri plates on the stage of a stereo-microscope equipped with a transmitted light source. Tricho-mycete thalli were teased out of the gut with fine-tippedforceps, and insect pins mounted in pin vises (Grobet USA,Carlstadt, New Jersey). Thalli were transferred to water on amicroscope slide (wet mounts), and examined for spores andother taxonomically significant characteristics with a com-pound microscope equipped with phase and differential in-terference contrast (Nomarski) optics. Semi-permanentvoucher slides were made by infiltrating specimens with adrop of lactophenol cotton blue stain (200 g phenol, 0.5 gcotton blue stain, 400 mL glycerol, 200 mL lactic acid, and200 mL distilled water) placed on the edge of the cover slip,then, after removing excess stain, sealing the edges of thecover slip with clear fingernail polish (Lichtwardt 1986;Lichtwardt et al. 2001a). Digital micrographs were taken ei-ther from living specimens (wet mounts) or stained, pre-served material on the voucher slides (read figure captionsto see which micrographs are from stained or fresh prepara-tions). Where possible, the species were identified using theLUCID keys for trichomycetes available at the University ofKansas Web site (Lichtwardt 2004). Type specimens for allnew species were deposited at the National MycologicalHerbarium, Agriculture and Agrifoods Canada, Ottawa, On-tario (DAOM). Additional voucher specimens of some ofthe new taxa, as well as other species reported, are availablefrom the author.

Results

Three new species of Harpellid trichomycetes, as well asa number of interesting previously described species, wereamong the trichomycetes collected from the guts of aquaticinsects from NL. The details are provided below.

Table 1. Description, collection date, and coordinates for sites in Newfoundland.

Site Description Collection date Coordinates and altitudeNL1 Goat cove (NF-26)*, pH 6.8, 6 8C 9 Nov 2006 47836’07@N, 052852’32@W; 79 mNL1 Goat cove (NF-26)*, no physical data 3 Jun 2007 47’836’07@N, 052852’32@W; 79 mNL2 Beachy cove (NF-2)*, no physical data 9 Nov 2006, 3 Jun 2007 Not recordedNL3 Beachy cove at Hugh’s pond outflow (NF-

1)*, pH 6.5, 7 8C9 Nov 2006 47835’56@N, 052850’56@W; 132 m

NL4 Broad cove river at outlet from Healey’spond (NF-19)*, pH 6.3, 7 8C

9 Nov 2006 47834’26@N, 052851’04@W; 145 m

GMH1 On highway (Hwy) 430 southeast ofGMMB towards Wiltondale, pH 6.3,12 8C

12 Sept 2008 49828’48@N, 057844’03@W; 7 m

GMH2 On Hwy 430 southeast of GMH1 towardsWiltondale, pH 7.5, 10 8C

12 Sept 2008 49828’15@N, 057842’59@W; 26 m

GMKL On road to Lomond (off highway 431) nearKilldevil Lodge, pH 8.0, 8 8C

12 Sept 2008 49829’49@N, 058802’17@W; 103 m

GMMB Mill Brook on Hwy 430 near DiscoveryCentre heading towards Wiltondale,pH 7.1, 11 8C

12 Sept 2008 49830’05@N, 057847’26@W; 10 m

GMTB Winterhouse Brook coming out of canyonat top of Tablelands trail, pH 7.8, 12 8C

10 Sept 2008 49828’03@N, 057857’23@W; 216 m

LOMR Lomond River under bridge on highway431, pH 8.0, 15 8C

10 Sept 2008 49824’08@N, 057843’48@W; 48 m

*Numbers in parentheses indicate the site codes used for these sites in Lichtwardt et al. (2001b).

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Figs. 1–6. Smittium colboi. Fig. 1. Sparse thallus with large trichospores (arrow), small trichospores (arrowheads), and a zygospore (arrowoutline). Fig. 2. Released zygospore with short collar (arrow). Figs. 3 and 4. Small trichospores. Fig. 5. Bulbous holdfast (arrow). Fig. 6.Large trichospores. Figs. 1, 5, and 6 are from lactophenol cotton blue stained specimens. Figs. 2, 3, and 4 were taken from unstained mate-rial. Scale bars = 20 mm.

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Taxonomy

Smittium colboi Strongman sp. nov. Figures 1–10Mycobank MB 518697

Thallus haptero bulboso inconspicuo exoriens (Fig. 5),sparsim ramosus; trichosporae dimorphae, trichosporismagnis 17.5–18 mm � 4–5.5 mm (Figs. 1, 6, 9) ellipsoideisin mediano tumidis, trichosporis parvis 10.5–14 mm � 2.5–3.5 mm (Figs. 1, 3, 4, 8) anguste ellipsoideis in mediano lev-iter tumidis; trichosporae ambae magnae parvaeque collum

breve expansum ferentes. Zygosporae biconicae, cylindricae50–59.5 mm � 4–5.5 mm (Figs. 1, 2, 7–10), typice in ex-tremo proximali leviter flexae (Figs. 7, 10), collo brevi postliberationem a zygosporophoro instructae. Zygosporopho-rum vel aggregatione communi basali cellularum ubi con-jungent aliquot rami (Fig. 10) vel directe ramo normaliexoriens. Ad parietem interiorem proctodaei larvarum Chi-ronomidarum affixus.

Thallus arising from an inconspicuous bulbous holdfast(Fig. 5), branching sparse, trichospores dimorphic with largetrichospores 17.5–18 mm � 4–5.5 mm (Figs. 1, 6, and 9), el-lipsoidal with a medial swelling, small spores 10.5–14 mm �

Figs. 7–10. Smittium colboi. Fig. 7. Zygospores borne on zygosporophores (arrow) arising from a hypha. Fig. 8. A zygospore (white arrow)and a small trichospore (arrow) attached to the same thallus. Fig. 9. A zygospore (arrowhead) and large trichospore (arrow) arising from thesame hypha. Fig. 10. Magnified basal section showing the zygosporophore and branch bearing large trichospores (arrows) arising from acommon mass of (empty) thallus cells (arrowhead). Figs. 9 and 10 are from lactophenol cotton blue stained specimens. Figs. 7 and 8 weretaken from unstained material. Scale bars = 20 mm.

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2.5–3.5 mm (Figs. 1, 3, 4, and 8), narrow-ellipsoidal with aslight medial swelling, both large and small trichosporeswith a short flared collar. Zygospores biconical, cylindrical,50–59.5 mm � 4–5.5 mm (Figs. 1, 2, and 7–10), typicallybent slightly at the end where the zygosporophore attaches(Figs. 7–10), with a short collar after release from the zygo-sporophore. The zygosporophore arises from either a com-munal basal aggregation of cells where a number ofbranches fuse (Fig. 10) or can form directly from an undif-ferentiated branch (Figs. 7 and 8). Attached to hindgut liningof midge (Chironomidae) larvae.ETYMOLOGY: The species is named in honour of Dr. MurrayColbo, an aquatic entomologist who has added extensivelyto the inventory of insect species found in NL, and who hashelped me and other trichomycetologists find insect hosts aswell as identify host species that contain new and interestingtrichomycetes.HOLOTYPE: Lactophenol cotton blue stained thalli, tricho-spores, and zygospores on microscope slide NL2–4 (DAOM241004), mount made from the hindgut of a chironomidlarva collected on 3 June 2007 at NL2 (Beachy Cove), nearSt. John’s.HABITAT: Dissected from the hindgut of chironomid (midge)larvae collected from a flowing stream at Beachy Cove,near St. John’s, NL.COMMENTARY: There are nine dimorphic Smittium spp. pub-lished (Lichtwardt 2004; Strongman et al. 2010). No otherdimorphic species has trichospore and zygospore dimensionsin the same range as S. colboi. The larger trichospores ofS. colboi (17.5 mm � 4–5.5 mm) are smaller than those ofany other dimorphic species. Smittium colboi has small tri-chospores (10.5–14 mm � 2.5–3.5 mm) that overlap inlength with five other species (Smittium biformeM.M.White & Lichtw., S. dimorphum Lichtw. &M.C. Williams, S. esteparum Ferrington, Lichtw. & Lopez

Lastra, S. orthocladii (Manier) Lichtw., and S. shanxienseJuan Wang, Strongman & S.Q. Xu) but the width is nar-rower in S. colboi compared with the width of small tri-chospores in these five species. There is overlap in thelength of zygospores of S. shanxiense (Strongman et al.2010) and S. colboi, but S. colboi zygospores are narrower.Smittium insulare Strongman has zygospores in the samesize range as S. colboi, but S. insulare is not dimorphic(Lichtwardt 2004).

Smittium georgense Strongman sp. nov. Figures 11–13Mycobank MB 518698Thallus densus, ramis numerosis haptero basali exorien-

tibus et abunde prolificantibus (Figs. 11, 12). Cellulae hap-teroideae basales initio nodosae (Fig. 12), aliquot hapterismassam magnam cum ramis radiantibus numerosis ultimoefferentibus (Figs. 11, 12). Trichosporeae subcylindraceae,34–45 mm � 5–7 mm (Fig. 13), collo 3–5 mm instructae.Zygosporae ignotae. Ad parietem interiorem proctodaei lar-varum Chironomidarum affixus.

Thallus dense with many branches arising from the basalholdfast and branching prolifically (Figs. 11 and 12). Hold-fast basal cells initially knobby (Fig. 12) with several hold-fasts ultimately forming a large mass with many radiatingbranches (Figs. 11 and 12). Trichospores sub-cylindrical,34–45 mm � 5–7 mm (Fig. 13), with a collar 3–5 mm, zygo-spores unknown. Attached to the hindgut lining of midge(Chironomidae) larvae.

ETYMOLOGY: Named after the well known and historicalGeorge Street in St. John’s, NL, not far from where this spe-cies was collected.

HOLOTYPE: Lactophenol cotton blue stained thalli and tricho-spores on microscope slide NL3–5 (DAOM 241002), mountmade from the hindgut of a chironomid larva collected on 3June 2007 at NL2 (Beachy Cove).

Figs. 11–13. Smittium georgense. Fig. 11. Mass of thallus branches arising from a central, knobby holdfast (arrow). Fig. 12. Young thalluswith multiple branches on a simple, knob-like holdfast (arrows). Fig. 13. Released trichospore. All figures are from unstained specimens.Scale bars = 20 mm.

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Figs. 14–16. Stachylina forantipes. Fig. 14. Thalli with attached trichospores (arrow). Fig. 15. Trichospores on a thallus with a bulbous footpenetrating the peritrophic membrane of the host. (arrow). Fig. 16. Released trichospore with single appendage (arrow) and no collar. Allfigures are from unstained specimens. Scale bars = 20 mm.

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HABITAT: Dissected from the hindgut of chironomid (midge)larvae collected from a flowing stream at Beachy Cove atthe outflow from Hugh’s Pond, near St. John’s, NL.

COMMENTARY: The trichospore size range found in S. geor-gense overlaps with numerous other species in the genus(Lichtwardt 2004; Siri and Lopez Lastra 2010), but onlythree species (Smittium cylindrosporum Lichtw. & Arenas,S. elongatum Lichtw., and S. rupestre Lichtw.) have cylin-drical or sub-cylindrical trichospores in this range and ashort collar like S. georgense (Lichtwardt 2004). Smittiumcylindrosporum has smaller trichospores on average, andboth this species and S. elongatum have verticilliate branch-ing of the thallus, which differs from S. georgense. The tri-chospores of S. rupestre are also slightly smaller thanS. georgense. Based on the current data available (Licht-wardt 2004; Wang et al. 2010), the distribution of bothS. rupestre (Australasian) and S. cylindrosporum (SouthAmerica) is very different from that for S. georgense.

Stachylina forantipes Strongman sp. nov. Figures 14–16Mycobank MB 518699

Thalli 35–50 mm � 6–9 mm, haptero parvo bulbosomembranam peritrophicam hospitis penetranti (Figs. 14,15). Trichosporae 2–4 in quoque thallo, ovoideae, 17.5–21 mm � 7–8 mm, in medio valde tumidae, sine collo(Fig. 16). Zygosporae ignotae. In membrana peritrophicalarvarum Chironomidarum.

Thalli 35–50 mm � 6–9 mm, with a small bulbous hold-fast penetrating the peritrophic membrane of the host(Figs. 14 and 15). Trichospores, 2–4 spores per thallus,

ovoid, 17.5–21 mm � 7–8 mm with a strong medial swelling,no collar (Fig. 16). Zygospores unknown. On peritrophicmembrane of midge (Chironomidae) larvae.ETYMOLOGY: From the Latin forare (to bore), and pes (foot),in reference to the penetrating holdfast.HOLOTYPE: Lactophenol cotton blue stained thalli and tricho-spores on microscope slide GMKL-8 (DAOM 241003),mount made from the peritrophic membrane of a chironomidlarva collected on 12 September 2008 from a small rockystream flowing through forest near Killdevil Lodge off high-way 431 in Gros Morne National Park, NL.HABITAT: Recovered from the peritrophic membrane of chiro-nomid (midge) larvae collected from a small, flowing streamnear Killdevil Lodge in the Gros Morne National Park area.COMMENTARY: There are two species that resemble S. foran-tipes and have thalli that penetrate the peritrophic membraneof the dipteran hosts. Stachylina minuta Gauthier ex Lichtw.has smaller trichospores 15–16 mm � 5–6 mm (Lichtwardt2004) than S. forantipes (17.5–21 mm � 7–8 mm). The thal-lus of S. minuta is narrower, and trichospores are producedon conspicuous outgrowths from the thallus (Lichtwardt2004), while those of S. forantipes are borne directly on thethallus. Stachylina minima M.C. Williams & Lichtw. tricho-spores (11–)14(–18) mm � 6–8 mm are also shorter, on aver-age, than those of S. forantipes. This species has a moresoutherly distribution than other species: New Zealand (Wil-liams and Lichtwardt 1990) and Argentina (Lopez Lastra etal. 2005).

Five of the eight species previously reported from NL,Harpella melusinae Leger & Duboscq and Pennella hovassi

Table 2. Trichomycetes recorded at each collection site with the host and collection date.

Species Site Host Collection dateGraminella microspora* GMKL Mayflies (Baetidae) 12 Sept 2008Harpella melusinae NL1, NL4 Black flies (Simuliidae) 9 Nov 2006; 3 Jun 2007Legeriomyces sp. NL3 Caddisflies (Limnophilidae) 9 Nov 2006Legeriomyces ramosus* GMMB, GMH1,

GMH2Mayflies (Ephemerellidae) 12 Sept 2008

Orphella avalonensis NL1 Stoneflies (Leuctridae) 9 Nov 2006Paramoebidium curvum NL1 Black flies 3 Jun 2007Paramoebidium spp. all Mayflies, black flies, caddisflies,

stoneflies9 Nov 2006; 3 Jun 2007;

10, 12, 13 Sept 2008Pennella hovassi NL1, NL2, NL4 Black flies 9 Nov 2006; 3 Jun 2007Pennella simulii NL2 Black flies 3 Jun 2007Smittium culicis* NL2 Midges (Chironomidae) 3 Jun 2007Smittium spp. GMKL, LOMR, NL1 Midges 9 Nov 2006; 10, 12 Sept

2008Smittium colboi sp. nov. NL2 Midges 3 Jun 2007Smittium georgense sp. nov. NL2 Midges 3 Jun 2007Spartiella barbata{ GMTB Mayflies 13 Sept 2008Spartiella sp. NL1 Mayflies 9 Nov 2006Stachylina grandispora* GMH1, LOMR Midges 10 Sept 2008Stachylina nana* NL2 Midges 3 Jun 2007Stachylina forantipes sp. nov. GMKL Midges 12 Sept 2008Stachylina sp. NL1, NL2 Midges 9 Nov 2006Zygopolaris ephemeridarum* GMKL Mayflies 12 Sept 2008Unidentified species GMTB Mayflies 10 Sept 2008

Note: Names in bold font are new taxa. For a full explanation of the site abbreviations see Table 1.*New records for Newfoundland.{New continental record.

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Manier ex Manier, in Frost and Manier (1971), as well asHarpella melusinae, Orphella avalonensis M.M. White,Lichtw. & Colbo, Paramoebidium curvum Lichtw., Pennellasimulii M.C. Williams & Lichtw., and Simuliomyces micro-sporus Lichtw., in Lichtwardt et al. (2001b), were also col-lected from the sites listed in Table 1. Harpellomycesabruptus Lichtw. M.M. White & Colbo, from a midge(Thaumeliidae) found in freshwater seeps, and Stachylina li-toralis Lichtw. M.M. White & Colbo, from a midge (Chiro-nomidae) that lives in the marine littoral habitat (Lichtwardtet al. 2001b) were not seen in my collections; however, nosamples were taken from these habitats.

Spartiella barbata was collected from mayflies in astream running out of the canyon on top of the Tablelands,Gros Morne National Park (GMTB, Table 2) and the charac-teristics of the NL collections match the species descriptionwell (Figs. 17–19). This species was previously known onlyfrom France (Manier 1962, 1968) and Spain (Valle 2007),therefore, this record from NL is a significant western rangeextension for the species. Three species, Graminella micro-spora S.T. Moss & Lichtw. (Figs. 23 and 24), Legeriomycesramosus Pouzar (Figs. 20 and 21), and Smittium culicisManier are new records for eastern Canada. Stachylina gran-dispora Lichtw., Stachylina nana Lichtw., and Zygopolarisephemeridarum S.T. Moss, Lichtw. & Manier (Fig. 22),were collected at the NL sites and have also been reportedfrom Prince Edward Island (PEI) (Strongman 2007). All sixof these species were reported from Ontario as well(Strongman and White 2008). The trichospores of Z. ephem-eridarum collected in NL were smaller on average (18.5–25.5 mm � 4.5–6 mm) than the holotype (25–38 mm � 5–8 mm), but all other characteristics were the same (Licht-wardt 2004).

Pennella species were common in the hindguts of blackfly larvae, where they were collected. Frost and Manier(1971) reported P. hovassi from collections of black fly lar-vae in NL, and the dimensions given coincide with the de-tails for specimens I collected at sites NL1, NL3, and NL4near St. John’s (Figs. 25–29). At one site (NL2), the thalliof P. hovassi were intermingled with thalli of P. simulii,making careful measurements of trichospores and zygo-spores necessary to determine which and how many specieswere present. Several species of Paramoebidium, as well asone or more species of Smittium and Stachylina were col-lected, but owing to the unavailability of mature thalli, iden-tification to species was not possible. Caddisfly larvae(Limophilidae) contained small obpyriform trichosporeswith two appendages on thalli in the hindgut (Table 2).There are no previously published species of Legeriomycesfrom these hosts, although I have retrieved several speciesfrom caddisfly larvae in collections from other provinces(D. Strongman, unpublished data). This collection did notyield sufficient specimens to give reliable size ranges for tri-

chospores. Furthermore, no zygospores were seen, so thespecimen was tentatively identified only as Legeriomycessp. Zygospores (Fig. 31) and one trichospore (Fig. 30) withcharacteristics seen in either Legeriomyces spp. or Legeriosi-milis spp. were seen in the hindguts of mayfly nymphs (Bae-tidae) at the Tablelands site (GMTB). No appendages wereseen on the trichospore. Members of these genera can bevery similar, except for the number of appendages on the tri-chospores (Lichtwardt 2004), so it was not possible to placethis specimen in a genus. It is recorded in Table 2 as an un-identified species.

DiscussionAlthough the diversity of aquatic insects on the island of

Newfoundland may be impoverished by comparison with themainland (Larson and Colbo 1983), the diversity of tricho-mycetes generally appears to be rich and their distributionis widespread. Seven species not previously reported fromNL are presented in this paper, and three new species aredescribed out of 17 species recovered at 10 sites. Of the 15trichomycetes that were identified to species, none werefound at both the St. John’s and Gros Morne locations(Table 2). This observation may be due mostly to the differ-ent collection dates, June and November in St. John’s andSeptember in Gros Morne, which is reflected in some differ-ences in the insect species composition. This is intriguingthough, and a more rigorous sampling on the east and westcoasts of the island during the summer and fall seasons mayreveal some interesting ecological information about theseareas that are separated geographically by over 500 km andarose from different geological origins.

The discovery of S. barbata in western NL is unexpected.The biogeographical theory whereby trichomycetes arethought to have migrated with their hosts, particularly non-vagile species like mayflies, and then speciated over timeafter the continents separated (Lichtwardt 1995) would notseem to support the movement of this species to westernNL, given that this region has a different geographical originthan the eastern part of the island (Owen 1991). However, itis possible that the fungus is also in the eastern region buthas not yet been discovered. Also, this species is found inEurope, most often in streams with high pH (Valle, personalcommunication), which was also true of the NL collections.Collections from high pH streams in other parts of NLmight show S. barbata to be more widely dispersed.

Pennella hovassi (the type species) described from blackfly larvae in France was reported in NL by Frost and Manier(1971), but Lichtwardt et al. (2001b) suggest that this recordis questionable because it was based on measurements thatcould have been distorted by the preservation of the materialexamined. They reported P. simulii from NL but not P. hov-assi (Lichtwardt et al. 2001b), and they noted slightlysmaller trichospore and zygospore size ranges in the NL col-

Figs. 17–24. Other trichomycetes collected from Newfoundland. Figs. 17–19. Spartiella barbata. Fig. 17. Trichospores. Note the com-pacted appendage in the sporogenous cell (arrow). Fig. 18. Zygospore. Fig. 19. Bulbous holdfast (arrow). Figs. 20 and 21. Legeriomycesramosus. Fig. 20. Obpyriform trichospore (arrowhead) with two appendages (arrows). Note appendages coiled in sporogenous cell in upperright. Fig. 21. Trichospore with 2 long appendages (arrows). Fig. 22. Zygospolaris ephemeridarum trichospores (arrows). Figs. 23 and 24.Graminella microspora. Fig. 23. New bulbous basal cells (arrows) formed at the ends of basal cells attached to the hindgut lining. Fig. 24.Trichospore with single appendage (arrow). All figures are from unstained specimens. Scale bars = 20 mm.

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Figs. 25–29. Pennella spp. Fig. 25. Trichospores of P. hovassi. Fig. 26. Holdfast of P. simulii. Fig. 27. Holdfast of P. hovassi. Fig. 28. Zygos-pores of P. hovassi. Fig. 29. Zygospore of P. simulii. All figures are from lactophenol cotton blue stained specimens. Scale bars = 20 mm.

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lections than for the holotype. I collected both of these spe-cies from black fly larvae at sites near St. John’s, NL1,NL2, and NL4 (Table 2). Pennella simulii was also collectedfrom NL2 and was intermingled with P. hovassi in my col-lections. These were sites also sampled by Lichtwardt et al.(2001b), but they were collected in April and May, while

my samples were collected in November and June. No col-lections of Pennella spp. were recorded from the GrosMorne sites. These collections were made in September andcontained no black fly larvae, so the larvae may have beentoo small to collect or the sites sampled did not have appro-priate habitat for this host.

There are eight species of Pennella and the main discrim-inating features are the dimensions of the trichospores andzygospores (Lichtwardt et al. 2001a). Description of theholdfast, particularly the nature of the branching of thisstructure, is important in differentiating the Pennella spp.from a similar harpellid also found in black fly larvae, Gen-istellospora homothallica Lichtw. Descriptions of the hold-fast in Pennella spp. range from simple, to bifurcated, todeeply and repeatedly bifurcated; however, since the hold-fast grows and divides as the thallus grows, it is not a reli-able feature for discriminating among Pennella spp. Table 3summarizes the features of the current Pennella spp. andshows that each can be accurately identified based on meas-urements of the mature trichospores and zygospores. Thereis substantial overlap in the trichospore dimensions forP. hovassi and Pennella digitata Strongman & M.M. White;however, the zygospores are much smaller in the latter, socorrect identification of each of these species is best donewhen both spore types are available. In Ontario we collectedboth P. digitata and P. simulii from the same black fly gut(Strongman and White 2008), while in Nova Scotia I haveseen P. hovassi, P. digitata, and P. simulii co-occurring inthe same collections (D. Strongman, unpublished data), andoften with more than one species in a single black fly gut.Careful observation is needed to determine which of thesespecies is present in black fly collections.

Zygopolaris ephemeridarum was collected in mayflynymphs from one site (GMKL) but the trichospores wereconsiderably shorter (18.5–25.5 mm � 4.5–6 mm) than themeasurements given for the holotype (25–38 mm � 5–8 mm) (Lichtwardt 2004). Collections of this species in PEI

Figs. 30–31. Unidentified trichomycete. Fig. 30. Released trichos-pore (stained with lactophenol cotton blue). No appendages seen.Fig. 31. Type II zygospores with a broad collar (arrow). Unstained.Scale bars = 20 mm.

Table 3. Morphometric data for Pennella species.*

Species Trichospores (mm) Zygospores (mm) Base

Species collected from NewfoundlandP. hovassi 23–33�4.5–7 68–79�7.5–13 Simply to deeply and repeatedly divided, bifur-

cate, mucilage, Type speciesP. simulii 30–41�6.5–10.5 84–96�19–24 Simple, tapered, occasionally digitate, mucilageP. simulii{ 28–38�7–10 74–89�16–19 Deeply divided, mucilage

Other speciesP. angustospora 50–104�2.5–6 82–92�15–17 Tapered to a rounded point, rarely slightly bifur-

cate, mucilageP. arctica 40–58�8–11 65–72�15–18 Simply to deeply and repeatedly divided, muci-

lageP. asymmetrica 35–52�12–17 Unknown Simple, mucilage, eccentric attachment of tri-

chospores on thallusP. digitata 27.5–33�6.5–8 (37)40–44�10–13.5 Deeply divided, bifurcate, mucilageP. grassei 49–59�4.5–5.5 90�15 Tapered, slightly digitate, mucilage unknown

(not described)P. montana 39–72�8–10 91–100�18–20 Dichotomusly bifurcated one or more times,

mucilage

*Data from Lichtwardt et al. 2001a{Modifications to spore size ranges from Newfoundland collections (Lichtwardt et al. 2001b)

Strongman 1021


(Strongman 2007) had trichospore and zygospore size rangesthat matched those given for the holotype. The trichosporesize range differences observed in the NL collection couldreflect geographic variation, however collections of the sex-ual spores from specimens in NL are needed to determinewhether the NL species is different from Z. ephemeridarum.

Given the unique geological features, abundance and diver-sity of aquatic systems, and the relative lack of information onthe biodiversity of trichomycetes from the island of New-foundland, future collections and studies of this group shouldbe informative and produce many more new species records.

AcknowledgmentsThanks to Carolyn Bird, Chester Grant, Nova Scotia, for

providing the Latin diagnoses and for invaluable feedbackon the species descriptions. Thanks also to Dr. MurrayColbo, Biology Department, Memorial University of New-foundland, who collected some of the samples for thiswork, identified some of the insects, and provided advicewith respect to collection sites in Gros Morne. I receivedpartial funding for this work from the Faculty of GraduateStudies, Saint Mary’s University.

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