A review and comparison of the nematode assemblages of the Australian golden bandicoot, Isoodon auratus, the quenda, I. fusciventer and southern brown bandicoot, I. obesulus (Peramelidae), from material held in the south Australian museum.

A total of 333 vials of nematodes collected from three species of Isoodon (representing three individuals of I. auratus, 63 of I. fusciventer and 92 of I. obesulus) held in the Australian Helminthological Collection of the South Australian Museum were examined. Nematodes were identified and the nematode assemblages of the three hosts were compared with each other and with the assemblage of Isoodon macrourus. Two fully identified species were recovered from I. auratus, eight from I. fusciventer and 14 from I. obesulus. None of the species occurred in all three hosts; Labiobulura inglisi (Subuluridae), Peramelistrongylus skedastos (Dromaeostrongylidae) and Asymmetracantha tasmaniensis (Mackerrastrongylidae) all occurred in I. fusciventer and I. obesulus. Only Pe. skedastos was also found in I. macrourus. Sorensen's index of similarity, 27.2 %, showed that I. fusciventer and I. obesulus did not have similar nematode communities and neither were their communities similar to that of I. macrourus, 17.1 % and 39.0 % respectively. Labiobulura inglisi and Linstowinema inglisi were the dominant nematodes in the assemblage of I. fusciventer and La. inglisi was dominant in I. obesulus. The two hosts had nematode assemblages with unique species profiles; one species of Linstowinema in I. fusciventer, three in I. obesulus; a species of Physaloptera in I. obesulus, none in I. fusciventer; four species of strongylid; Asymmetracantha tasmaniensis the most prevalent in I. fusciventer, Peramelistrongylus skedastos the most prevalent in I.obesulus. The size of the geographic range is a probable determinant of the species richness of the nematode assemblages.

A review of the nematode assemblage of the Australian bandicoot, Isoodon macrourus (Peramelidae), from material held in the South Australian Museum with the description of Sprattellus cassonei n. sp. (Mackerrastrongylidae).

A total of 235 vials of nematodes held in the Australian Helminthological Collection of the South Australian Museum from 125 individuals of Isoodon macrourus were examined. The nematode assemblage of I. macrourus, comprising 12 families, including 16 genera and 23 identified species, was compared with the sympatric bandicoot species Perameles nasuta, 20 identified species (Sorensen's index of similarity 0.56) and P. pallescens, 12 identified species (Sorensen's index 0.51). Sprattellus cassonei n. sp. is distinguished from its congeners by having a synlophe with 7-8 ridges with the anterior ventral ridges interrupted, the morphology of the dorsal ray and the branching of the spicule tips. A single male specimen identified as Linstowinema sp. 1. is characterised by seven circles of body hooks, the oesophagus terminating at the level of the seventh circle and robust scale-like spines on the posterior ventral body. A complete description of the species will require additional material, including females. Difficulties in identifying individuals of the genus Mackerrastrongylus to species level are discussed. Overall similarities in the nematode assemblages of the three bandicoot hosts are likely due to shared relationships and similar behaviours.

Experimental infections, using a fluorescent marker, of two elasmobranch species by unciliated larvae of Branchotenthes octohamatus (Monogenea: Hexabothriidae): invasion route, host specificity and post-larval development.

The infection biology of Branchotenthes octohamatus (Monogenea: Hexabothriidae) from the gills of the southern fiddler ray, Trygonorrhina fasciata (Rhinobatidae), was studied using the fluorescent dye, 5(6)-carboxyfluorescein diacetate N-succinimidyl ester (CFSE). This is the first use of this technique on a monogenean species with unciliated larvae and the first for any monogenean larva infecting an elasmobranch host. CFSE-labelled post-larvae were recovered from gills of T. fasciata within 30 min of exposure to the host, providing strong evidence that larvae invade host gills directly and do not migrate after initial attachment elsewhere. The rapidity with which larvae settled suggests that the mode of infection may deliver larvae directly to the gills via the host's inhalant respiratory current. The specificity of B. octohamatus was investigated by exposing a sympatric rhinobatid host species, the western shovelnose ray, Aptychotrema vincentiana, to B. octohamatus larvae newly emerged from eggs laid by adult parasites from gills of T. fasciata. Experimental exposure of A. vincentiana to freshly hatched B. octohamatus larvae resulted in a persistent infection, indicating that B. octohamatus may not be strictly host specific. Post-larval development charted on these experimentally infected A. vincentiana specimens was slow. Parasites appeared to be sexually mature at 91 days at 21-25 degrees C. Branchotenthes octohamatus larvae bear only 4 pairs of hooklets on the haptor whereas all other hexabothriid larvae described so far have 5 hooklet pairs. Ontogenetic changes to the haptor revealed that it is probably hooklet pair III that is lost from B. octohamatus prior to larval development.

The oncomiracidium of Nasicola klawei (Monogenea: Capasalidae: Capsalinae).

The oncomiracidium of the capsaline Nasicola klawei (Monogenea: Capsalidae), adults of which were collected from the nares of the yellowfin tuna Thunnus albacares (Scombridae) caught off Nelson Bay on the east coast of Australia, is described. This is a new locality record for N. klawei. This is the first description of an oncomiracidium of a Nasicola species and only the third time the larva of a capsaline species has been documented. The oncomiracidium of N. klawei is distinguished from others in the subfamily by the distribution of the body pigment and large distinct refringent (lipid?) droplets in the body and the prominent, posteriorly directed pointed process on the guard of each of the 14 hooklets. Oncomiracidia of the other two described Nasicola species, N. hogansi and N. brasiliensis, should be examined to determine whether these characters have generic or species-specific significance.

Three unrelated species, 3 sites, same host--monogenean parasites of the southern fiddler ray, Trygonorrhina fasciata, in South Australia: egg hatching strategies and larval behaviour.

The southern fiddler ray, Trygonorrhina fasciata (Rhinobatidae), is parasitized by 3 monogenean (platyhelminth) species from 3 families on 3 different sites of the host: Calicotyle australis (Monocotylidae) from the cloaca, Pseudoleptobothrium aptychotremae (Microbothriidae) from the skin and Branchotenthes octohamatus (Hexabothriidae) from the gills. Cues that promote egg hatching were investigated for each species and the behaviour of their larvae was also documented. Eggs were laid by parasites in vivo and maintained at 22 degrees C. Three different egg hatching and host finding strategies were discovered. Calicotyle australis eggs hatched spontaneously with a strong diurnal rhythm that is likely to be under circadian control. The larva is ciliated, photo-responsive and can survive for up to 24 h at 22 degrees C after hatching. Pseudoleptobothrium aptychotremae may have a 'bet-hedging' strategy. Some eggs hatched spontaneously and rhythmically. However, since the hatching success was low, it is possible that other eggs require a different cue provided by the host. The larva is also ciliated but shows no photo-response and was observed to remain active for <4 h at 22 degrees C after hatching. Branchotenthes octohamatus has a 'sit-and-wait' strategy that depends on mechanical disturbance to stimulate hatching. The larva is unciliated, shows no photo-response but may survive for more than 2 days at 22 degrees C after hatching. The implications of hatching strategy, larval behaviour and morphology in the goal to find a host are discussed for each species.

A preliminary phylogenetic analysis of the Capsalidae (Platyhelminthes: Monogenea: Monopisthocotylea) inferred from large subunit rDNA sequences.

Phylogenetic relationships within the Capsalidae (Monogenea) were examined using large subunit ribosomal DNA sequences from 17 capsalid species (representing 7 genera, 5 subfamilies), 2 outgroup taxa (Monocotylidae) plus Udonella caligorum (Udonellidae). Trees were constructed using maximum likelihood, minimum evolution and maximum parsimony algorithms. An initial tree, generated from sequences 315 bases long, suggests that Capsalinae, Encotyllabinae, Entobdellinae and Trochopodinae are monophyletic, but that Benedeniinae is paraphyletic. Analyses indicate that Neobenedenia, currently in the Benedeniinae, should perhaps be placed in a separate subfamily. An additional analysis was made which omitted 3 capsalid taxa (for which only short sequences were available) and all outgroup taxa because of alignment difficulties. Sequence length increased to 693 bases and good branch support was achieved. The Benedeniinae was again paraphyletic. Higher-level classification of the Capsalidae, evolution of the Entobdellinae and issues of species identity in Neobenedenia are discussed.

A comparison of the anterior adhesive system in the oncomiracidium and adult of the monogenean parasite Menizocotyle icopae (Monocotylidae).

The anterior adhesive system of the oncomiracidium and adult of Merizocotyle icopae (Monogenea: Monocotylidae) were compared. The oncomiracidium has one ventrally placed aperture on either side of the head near the anterior extremity. In the adult, there are three ventrally placed apertures on either side of the head region. Both systems have three types of electron-dense secretory bodies opening into each aperture. A rod-shaped secretion (S1) and a small electron dense ovoid secretion (S2) are common to larvae and adults. The third secretion type differs: in adults, it is a large, spherical (S3) type but in larvae, it is an ovoid (S4) body. S4 bodies do occur in adults, but appear to be secreted as a general body secretion. An additional anteromedian secretion (S5) is also present in the oncomiracidium, but is not secreted into the anterior apertures. Homology and function of secretions are discussed.

Invasion of the shovelnose ray (Rhinobatos typus) by Neoheterocotyle rhinobatidis and Merizocotyle icopae (Monogenea: Monocotylidae).

This study examined the route of infection by free-swimming larvae of 2 monocotylid monogeneans that inhabit the gills (Neoheterocotyle rhinobatidis) and the nasal tissue (Merizocotyle icopae) of the shovelnose ray, Rhinobatos typus, from Heron Island on the Great Barrier Reef, Australia. Larvae of N. rhinobatidis and M. icopae attached directly to the gills and the nasal tissue of the ray, respectively, and did not first settle on the skin. Initial development of the post-oncomiracidium of N. rhinobatidis was rapid and hamuli formed between 6 and 24 h p.i. at a mean temperature of 26 degrees C. However, growth then slowed markedly and was variable; only 2 fully mature individuals were found 20 days p.i. at a mean temperature of 24.5 degrees C. Development of M. icopae was slow and variable throughout; hamuli did not appear until 10 days p.i. and no mature individuals were obtained even 22 days p.i. at a mean temperature of 24.5 degrees C. No character could be found as an indicator of parasite age for N. rhinobatidis or M. icopae due to the high variability in development in both species.

First published record of the pathogenic monogenean parasite Neobenedenia melleni (Capsalidae) from Australia.

The monogenean Neobenedenia melleni (MacCallum, 1927) Yamaguti 1963 is a well-known and virulent pathogen in culture conditions recorded from the skin of many teleost fish species worldwide. Until now, N. melleni has not been reported from wild or cultured fish in Australian waters. This study documents a recent outbreak of N. melleni that occurred on Lates calcarifer (barramundi) cultivated in sea cages in Hinchinbrook Channel between Hinchinbrook Island and mainland Queensland, Australia, which resulted in the loss of 200,000 fish (50 tonnes). The origin of this outbreak is unclear because N. melleni has not been recorded from any wild host species in Australia and strict quarantine regulations exclude the possibility of its introduction on imported fish. We propose that N. melleni occurs naturally on wild populations of some teleost species in Australian waters and that the few surveys of wild fish conducted along the east coast have failed to report this species. The possibility that uncharacteristically low water temperatures led to the outbreak is discussed.

Phylogenetic analysis of the Monocotylidae (Monogenea) inferred from 28S rDNA sequences.

The current classification of the Monocotylidae (Monogenea) is based on a phylogeny generated from morphological characters. The present study tests the morphological phylogenetic hypothesis using molecular methods. Sequences from domains C2 and D1 and the partial domains C1 and D2 from the 28S rDNA gene for 26 species of monocotylids from six of the seven subfamilies were used. Trees were generated using maximum parsimony, neighbour joining and maximum likelihood algorithms. The maximum parsimony tree, with branches showing less than 70% bootstrap support collapsed, had a topology identical to that obtained using the maximum likelihood analysis. The neighbour joining tree, with branches showing less than 70% support collapsed, differed only in its placement of Heterocotyle capricornensis as the sister group to the Decacotylinae clade. The molecular tree largely supports the subfamilies established using morphological characters. Differences are primarily how the subfamilies are related to each other. The monophyly of the Calicotylinae and Merizocotylinae and their sister group relationship is supported by high bootstrap values in all three methods, but relationships within the Merizocotylinae are unclear. Merizocotyle is paraphyletic and our data suggest that Mycteronastes and Thaumatocotyle, which were synonymized with Merizocotyle after the morphological cladistic analysis, should perhaps be resurrected as valid genera. The monophyly of the Monocotylinae and Decacotylinae is also supported by high bootstrap values. The Decacotylinae, which was considered previously to be the sister group to the Calicotylinae plus Merizocotylinae, is grouped in an unresolved polychotomy with the Monocotylinae and members of the Heterocotylinae. According to our molecular data, the Heterocotylinae is paraphyletic. Molecular data support a sister group relationship between Troglocephalus rhinobatidis and Neoheterocotyle rhinobatidis to the exclusion of the other species of Neoheterocotyle and recognition of Troglocephalus renders Neoheterocotyle paraphyletic. We propose Troglocephalus incertae sedis. An updated classification and full species list of the Monocotylidae is provided.

Phylogenetic analysis of the Monocotylidae (Monogenea) inferred from 28S rDNA sequences.

The current classification of the Monocotylidae (Monogenea) is based on a phylogeny generated from morphological characters. The present study tests the morphological phylogenetic hypothesis using molecular methods. Sequences from domains C2 and D1 and the partial domains C1 and D2 from the 28S rDNA gene for 26 species of monocotylids from six of the seven subfamilies were used. Trees were generated using maximum parsimony, neighbour joining and maximum likelihood algorithms. The maximum parsimony tree, with branches showing less than 70% bootstrap support collapsed, had a topology identical to that obtained using the maximum likelihood analysis. The neighbour joining tree, with branches showing less than 70% support collapsed, differed only in its placement of Heterocotyle capricornensis as the sister group to the Decacotylinae clade. The molecular tree largely supports the subfamilies established using morphological characters. Differences are primarily how the subfamilies are related to each other. The monophyly of the Calicotylinae and Merizocotylinae and their sister group relationship is supported by high bootstrap values in all three methods, but relationships within the Merizocotylinae are unclear. Merizocotyle is paraphyletic and our data suggest that Mycteronastes and Thaumatocotyle, which were synonymized with Merizocotyle after the morphological cladistic analysis, should perhaps be resurrected as valid genera. The monophyly of the Monocotylinae and Decacotylinae is also supported by high bootstrap values. The Decacotylinae, which was considered previously to be the sister group to the Calicotylinae plus Merizocotylinae, is grouped in an unresolved polychotomy with the Monocotylinae and members of the Heterocotylinae. According to our molecular data, the Heterocotylinae is paraphyletic. Molecular data support a sister group relationship between Troglocephalus rhinobatidis and Neoheterocotyle rhinobatidis to the exclusion of the other species of Neoheterocotyle and recognition of Troglocephalus renders Neoheterocotyle paraphyletic. We propose Troglocephalus incertae sedis. An updated classification and full species list of the Monocotylidae is provided.

The Calicotyle conundrum: do molecules reveal more than morphology?

Partial large subunit 28S rDNA sequences were obtained for specimens of Calicotyle (Monogenea: Monocotylidae) from eight different host species distributed worldwide to test the validity of some species and to address the question of host-specificity in others. Sequences obtained for Calicotyle specimens identified as C. kroyeri based on morphological methods from the type-host Raja radiata (Rajidae) and an additional host R. clavata, both from the North Sea, were identical. However, 'C. kroyeri' from the cloaca of R. naevus from Tunisia, Raja sp. A from Tasmania and R. radula from Tunisia differed from C. kroyeri from R. radiata by five (0.51%), 21 (2.13%) and 39 (3.96%) base pairs, respectively, over 984 sites. Therefore, it is likely that the specimens from Raja sp. A, R. radula and perhaps even from R. naevus are not C. kroyeri. Molecular results determined that the calicotylines from the cloaca of Urolophus cruciatus and U. paucimaculatus (Urolophidae) from southern Tasmania identified previously as C. urolophi are indeed identical. Large subunit 28S rDNA sequences of C. palombi and C. stossichi collected from the cloaca and rectal gland, respectively of Mustelus mustelus (Triakidae) from the coast of Tunisia differ sufficiently for these calicotylines to be considered separate and valid species. Our results indicate that some species of Calicotyle are not strictly host-specific, but that C. kroyeri may not be as widely distributed in rajids as was believed previously. Calicotyle specimens from rajids must be re-examined critically to determine whether there are morphological differences indicative of specific differences that may have been overlooked previously.

Euzetia occultum n. g., n. sp. (Euzetiinae n. subf.), a monocotylid monogenean from the gills of Rhinoptera neglecta (Rhinopteridae) from Moreton Bay, Queensland, Australia.

Euzetia occultum n. g., n. sp. (Monogenea: Monocotylidae) is described from the gills of the Australian cownose ray Rhinoptera neglecta Ogilby collected in Moreton Bay, Queensland, Australia. Euzetia has one central and ten peripheral loculi, which is similar to species in Decacotyle Young, 1967. However Euzetia is distinguished from other genera in the family by the presence of an additional loculus on either side of the central loculus. Because Euzetia does not fit into any of the six existing subfamilies in the Monocotylidae Taschenberg, 1879, as currently recognised, we propose the Euzetiinae n. subf. to accommodate the new genus. Euzetia occaltum is described and illustrated fully. This is the first published record of a monocotylid from a species of Rhinoptera Cuvier.

Dendromonocotyle colorni sp. n. (Monogenea: Monocotylidae) from the skin of Himantura uarnak (Dasyatididae) from Israel and a new host record for D. octodiscus from the Bahamas.

Dendromonocotyle colorni sp. n. (Monogenea: Monocotylidae) is described from the dorsal skin surface of two specimens of Himantura uarnak (Forsskål) kept at the Eilat Underwater Observatory in Israel. Dendromonocotyle colorni is distinguished from the other eight species in the genus by the morphology of the terminal papillar sclerite on the haptor, the distal portion of the male copulatory organ and the morphology of the vagina. The development of the male copulatory organ is detailed for D. colorni and the adaptations of species of Dendromonocotyle to life on the dorsal skin surface of rays are discussed. Dendromonocotyle octodiscus Hargis, 1955 was identified from the dorsal skin surface of the southern stingray Dasyatis americana Hildebrand et Schroeder off Bimini, Bahamas and represents a new host record.

Egg hatching in 3 species of monocotylid monogenean parasites from the shovelnose ray Rhinobatos typus at Heron Island, Australia.

Eggs of Neoheterocotyle rhinobatidis, Troglocephalus rhinobatidis and Merizocotyle icopae (Monogenea: Monocotylidae) from the shovelnose ray Rhinobatos typus (Rhinobatidae) have a distinct hatching pattern linked to light periodicity. Larvae of these 3 monogenean species emerge only during daylight when exposed to natural illumination or when incubated in alternating 12 h periods of light and dark (light on 06.00 h, light off 18.00 h). N. rhinobatidis larvae emerge with a distinct peak during the first 2 h of light; this peak is not as pronounced in T. rhinobatidis or M. icopae. Eggs of N. rhinobatidis incubated in a reverse light/dark cycle (light on 18.00 h, light off 06.00 h) hatched only during periods of illumination, again with a peak during the first 2 h of light. Evidence suggests that the hatching patterns observed in all 3 species represent true circadian rhythms because eggs incubated in 24 h light or 24 h dark conditions continued to hatch with a rhythm. Shadows, disturbance and host tissue did not promote hatching in N. rhinobatidis or T. rhinobatidis but there were indications that host tissue may promote hatching in M. icopae. The hatching patterns observed are discussed with respect to their adaptive responses to host behaviour and predation pressure.

Heliocotyle ewingi sp. n. (Monogenea: Monocotylidae) from the gills of Myliobatis australis (Myliobatididae) from Tasmania, Australia.

Heliocotyle ewingi sp. n. (Monogenea: Monocotylidae) is described from the gills of Myliobatis australis Macleay, 1881 (Myliobatididae) collected from Norfolk Bay near Hobart, Tasmania, Australia. Heliocotyle ewingi can be distinguished readily from the only other species in the genus, Heliocotyle kartasi Neifar, Euzet et Ben Hassine, 1999, by the presence of a single pseudoseptum on each of the peripheral loculi except the posteriormost, eyespots and by the morphology of the male copulatory organ which is a short, straight sclerotised tube which lacks a sclerotised accessory piece. The generic diagnosis is revised to accommodate the new species and the anterior glands are discussed.

A new species of Neoheterocotyle Hargis, 1955 (Monogenea: Monocotylidae) from the gills of Pristis clavata Garman (Pristidae) from Darwin, Australia.

Neoheterocotyle darwinensis n. sp. is described from between the secondary gill lamellae of the dwarf sawfish Pristis clavata Garman (Pristidae) collected at the mouth of Buffalo Creek near Darwin, Northern Territory, Australia. This is only the second monocotylid species to be described from northern Australia. N. darwinensis is distinguished from the other seven valid species in the genus by the morphology of the hamuli, the dorsal haptoral accessory sclerites and the male copulatory organ. The similarities between N. darwinensis and Nonacotyle pristis Ogawa, 1991 from the gills of the freshwater sawfish Pristis microdon Latham collected in Papua New Guinea are discussed.

Ultrastructure of the eyes of the larva of Neoheterocotyle rhinobatidis (Platyhelminthes, Monopisthocotylea), and phylogenetic implications.

The oncomiracidium of Neoheterocotyle rhinobatidis (Monogenea, Monopisthocotylea, Monocotylidae) has two pairs of eyes, each eye with a lens and pigment cup. The anterior eyes have a single rhadomere; the posterior ones, two rhadomeres. Lenses are part of the pigment cup cells, as indicated by cytoplasmic connections between them and the pigment cups, and they are of mitochondrial origin because mitochondrial cristae are present in the periphery of the lenses. This is the first time that mitochondrial lenses have been shown to exist in a neodermatan. Such lenses may be a synapomorphy of a large taxon comprising the Neodermata and its turbellarian sister groups, or they may have evolved convergently in several not closely related groups as a result of strong selection pressure to find a suitable habitat or host.

Observations on the ultrastructure of the anterior adhesive areas and other anterior gland cells in the monogenean Merizocotyle australensis (Monocotylidae) from the nasal fossae of Himantura fai (Dasyatididae).

The anterior adhesive areas of a monocotylid monogenean, Merizocotyle australensis (Merizocotylinae), were investigated. They comprise 6 ventral apertures in 2 groups of 3 arranged at the anterolateral margins of the head. These regions are also well supplied with groups of cilia. Each aperture is 13.8 to 15.8 microm wide and contains multiple tubular projections that are covered with microvilli through which open 2 types of secretory cell ducts that carry either rod-shaped or spherical secretory bodies. The gland cell bodies that produce these 2 types of secretions co-occur at the anterior end. The 2 types of secretory bodies occur adjacent to one another and both are present in the extruded adhesive. The membranes of rod-shaped bodies are retained in the extruded glue. Rod-shaped bodies are 390 +/- 18 nm wide, at least 10.9 microm long, and show 2 types of internal periodic banding: 10.6 nm and 143 +/- 3 nm. The spherical vesicles are 130 +/- 6 nm in diameter and are electron-dense. A third secretion is present in separate ducts that also open anteriorly but emerge through the tegument between the ventral apertures. This secretion does not appear to be part of the adhesive secretion. The bodies of the third secretion are elongate, electron-dense, and 374 +/- 23 nm long. Inside the "lip" of the aperture, general body tegument abuts tegument specific to the aperture. The general body tegument is thicker, contains electron-dense vesicles, and has a ridged surface devoid of microvilli. Where the 2 kinds of tegument meet, they are connected by septate desmosomes.

Ciliated cells and chaetotaxy of the larvae of seven species of monocotylid monogeneans (Platyhelminthes) from Heron Island, Great Barrier Reef, Australia.

The distribution and the number of ciliated cells and sensilla of the oncomiracidia of seven species representing five of the six subfamilies of monocotylid monogeneans are described. The species examined include Troglocephalus rhinobatidis Young, 1967 from the Dasybatotreminae; Decacotyle floridana (Pratt, 1910) Chisholm and Whittington, 1998 from the Decacotylinae; Heterocotyle capricornesis Chisholm and Whittington, 1996 and Neoheterocotyle rhinobatidis (Young, 1967) Chisholm, 1994 from the Heterocotylinae; Merizocotyle icopae Beverley-Burton and Williams, 1989 from the Merizocotylinae; and Clemacotyle australis Young, 1967 and Monocotyle spiremae Measures, Beverley-Burton and Williams, 1990 from the Monocotylinae. The number of ciliated cells in each zone (anterior, median and haptoral) is similar between the species, but the distribution of these cells varies and generally supports the subfamilial divisions proposed for the Monocotylidae on the basis of a phylogenetic analysis of morphological characters of the adults. A silver-staining technique modified from the method of Lambert is described.