© Institute of Parasitology, Biology Centre CAS >;+'-F;+-& doi: '+'99''G;+'-+-& http://folia.paru.cas.cz

Research Article Morphological and molecular data for three of the (: ) in Australia, brevisacciferum Shimazu et Pearson, 1991 and minutus Johnston, 1948

Olena Kudlai';&, Scott C. Cutmore4 and Thomas H. Cribb4

1!5$ D5#67= ; JK#M$= 3 Q7R6UQ= 4 The University of Queensland, School of Biological Sciences, Brisbane, Queensland, Australia

Abstract: Cercariae and metacercariae of three species of the Microphallidae Y#'Z;+were found in snails and crustaceans 776#[!Maritrema brevisacciferum Shimazu et Pearson, 1991 and Microphallus minutus ]'Z9<, which have previously been reported in Queensland, were found as cercariae in the tateid gastropod Posticobia brazieri (Smith) and as metacercariae of M. brevisacciferum in the atyid shrimp Caridina indistincta Calman and of M. minutus[Cherax dispar 6$. Combined analysis of morphological and molecular data, based on newly Y!;;

Y _ Y# 'Z;+ However, despite the numerous records of microphal@ globally distributed family of digeneans that parasitise [#` #7 xJ7$ ;++<{ larval stages from intermediate hosts from this region. but have their greatest richness in birds and mammals. _ '; # 7 @ The life cycles of microphallids involve gastropods, most@ =77$@ 7$ ` [ ish water snail, Coxiella badgerensis Johnston (see John@ and crustaceans as second intermediate hosts. Previous 'Z9<J7$'ZF<'ZFZ!'Z*9 studies from Australia have reported 35 microphallid spe@ 'Z<& J7$ 'ZZ+ ! 'ZZ' ';xAtriophallophorusJ7$6 J7$'ZZF{Y@ 'ZF9=Basantisia'Z&<=Endocotyle Belopol'skaya, `7x'Z*<{`@ 'Z-;= Gynaecotyla ~ 'Z&Z= Levinseniella Stiles [  'Z+'= Maritrema R 'Z+*= Microphallus cerithiid gastropod, Clypeomorus batillariaeformis Habe € 'Z+'= Mochliscotrema J7$ 'Z

!#$&'&*++-5$ D5#6789;+&<***-9&*=>?89;+@&<@-&'+&<<=B@$D

This is an Open Access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/4.0), which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited. '+'99''G;+'-+-& >`icrophallid larvae in Australia

Table 1. Host data, geographical origin and GenBank accession numbers for digenean species used in phylogenetic analyses.

Species Host Country GenBank 6 N;<;F;9* @x;+''{ (Sowerby) M. heardi (Kinsella et Deblock, 1994) Oryzomys palustris (Harlan) >M! ~;;+F&; Y$x;++&{ M. neomiY$'ZZ< Neomys anomalus Cabrera K$$6M$ >'-'Z;* Y$x;+++{ M. novaezealandense Martorelli, Zeacumantus subcarinatus Portobello Bay, Dunedin, New ]'99'*< Presswellx;+'9{ >7_et;++9 (Sowerby) Zealand M. oocysta (Lebour, 1907) Hydrobia ulvae (Pennant) Belfast Lough, Northen Ireland ~;;+F&+ Olsonx;+03) M. poulini`@ Paracorophium excavatum Lake Waihola, Waihola, New KJ144177 Presswellx;+'9{ et#;+'9 (Thomson) Zealand M. prosthometra Deblock et Heard, 1970 Oryzomys palustris >M! ~;;+F&' Y$x;++&{ M. subdolum Jägerskiöld, 1909 Hydrobia ulvae Kandalaksha Bay, White Sea, _-<9'&- $#x;+';{ 6 M. deblocki`@ Anas platyrhynchos Linnaeus B New KJ144173 Presswellx;+'9{ et #;+'9 Zealand Microphallus Ward, 1901 M. abortivus Deblock, 1974 Hydrobia ulvae Belfast Lough, Northen Ireland ~;;+F;F Y$x;++&) M. basodactylophallusx'ZFZ{ Oryzomys palustris >M! ~;;+F;< Y$x;++&{ M. fusiformis6'ZF& Hydrobia ulvae Belfast Lough, Northen Ireland ~;;+F&& Y$x;++&{ M. minutus ]'Z9< Posticobia brazieri Brisbane, Australia Y&--<;& Present study M. minutus Cherax dispar6$ Brisbane, Australia Y&--<;; Present study M. primasx]$'Z+<{ Hydrobia ulvae Belfast Lough, Northen Ireland ~;;+F;* Y$x;++&{ M. similis (Jägerskiöld, 1900) Carcinus maenas (Linnaeus) Belfast Lough, Northen Ireland ~;;+F;- Y$x;++&{ M. triangulatus $#'Z<9 Somateria mollissima Bona@ Yamskaya Bay, Sea of Okhotsk, _-<9'&Z $#x;+';{ parte et Gray 6 Microphallus sp. Somateria mollissima Cape Taygonos, Sea of _-<9'9; $#x;+';{ $$6 Microphallus sp. Austrolittorina cincta (Quoy et €R6$, New Zealand ]

MATERIALS AND METHODS x {R`Kx;+';` ;+'9{ Sample collection During a study of parasites infecting freshwater inverte@ ! ` >7@ 776#7 ;+'- 7 7 6# of the tateid gastropod Posticobia brazieri (Smith) and the _ $ x;* &+‰'F‰‰!= '-; --‰-+‰‰B{ 7$ € crustaceans Caridina indistincta Calman and Cherax dis- x;* 9F‰'F‰‰!= '-; 9'‰+;‰‰B{ > $ x;* ;<‰-<‰‰!= par 6$`7`# '-; 9+‰+9‰‰B{ 99<+ Posticobia brazieri ` ?@ stages of species belonging to the family Microphallidae. &< Caridina indistincta, 34 Macrobrachium Detailed morphological study and comparative analyses tolmerum6$Cherax dispar`?_@ 7 `@7 j 7@ gill Creek and Churchbank Weir. Y!;;

>;+'-F;+-& ;'& '+'99''G;+'-+-& >`icrophallid larvae in Australia with SPOT™ imaging software. All measurements are in micro@ was then trimmed to the length of the shortest sequence. Phy@ metres and are given as a range followed by the mean and the logenetic analysis was carried out using Bayesian inference (BI) number of measurements taken (n) in parentheses. Drawings are _?Q$x_Q{Y7@ based on digital images of live cercariae and metacercariae. tution model, the general time reversible model, with estimates of #7@# Molecular data x Y688 {`5_Y;`xJ@ Total genomic DNA was isolated using phenol/chloroform 7;+';{`_@ ?jx!7$6;++'{Y `x#&;&{x6j;+';{6B! J'MJ&;Y#'9`x67;+';{ Z- 9`7&+ Z- '-F '*; RESULTS ?;`7Z- @ B?99<+#Posticobia brazie- '-- 9-[*; ? ri 76#7 9Y`[`Y!; Queensland revealed infections with digeneans from eight Z- & families. Nine infections consistent with species of the 9- ;*; ?Z+`7 _`[7 9Z- 9--+ 9- emerged cercariae. Metacercariae of Maritrema brevisac- *; ?Z+`7&+Z- @ ciferum and Microphallidae gen. sp. were found in Carid- ;+-; ;+*; ?Z+ ina indistincta= metacercariae of Microphallidae gen. sp. `7[*; ?- were also found in huge numbers in the tissues of a single [ JR ` [ !QYB P. brazieri. Metacercariae of Microphallus minutus were 6 R@ recovered from Cherax dispar. No specimen of Macrobra- j[JR` chium tolmerum was infected by digenean metacercariae. Dye™ v.3.1 chemistry following the manufacturer’s recommen@ Descriptions of the mature cercariae and metacercariae of 6[ these digeneans are provided below. `;x-‰@ Y@ Y YY@&‰= Q` ;+++{ BJ; > Microphallidae Y#'Z;+ x-‰@YY Y Y YYY @&‰= Q` ;+++{Y!; 'x-‰@ Y @ Genus Maritrema Nicoll, 1907 YYY @&‰=$'ZZ<{j@ ` 6> Maritrema brevisacciferum Shimazu et Pearson, 1991 &*&+? j !j( # >' 4.5 (GeneCodes Corp.) was used to assemble and edit contiguous j $ 7 ? j Description of cercaria x7 ;+ # this study are shown in Table 1. emerged specimens from P. brazieri from Moggill Creek): R`;

>;+'-F;+-& Page 3 of 13 '+'99''G;+'-+-& >`icrophallid larvae in Australia

Fig. 1. Maritrema brevisacciferum Shimazu et Pearson, 1991. A – cercaria from Posticobia brazierix!{##`=B – stylet, ##`=C – metacercaria from Cardina indistincta Calman.

[with secretory material in form of small granules, lo@ 77 = 77 # j7-;MZ+xFF=š'<{ B?#~@= ?7 @#=. @7#J#@ > Posticobia brazieri (Smith) let. Genital primordium an oval mass of cells in posterior (: ). 7B?#~@>@@ Locality: _ $ > $ [ ;x;8;{8x;8;{žš'F Australia. Description of metacercaria x7;<#@ Prevalence of emergence: ';'F+xMoggill Creek), ed specimens from _$M'<Churchbank *;+<+x>${ €M'+{_@#'F'M;'-& Second intermediate host: Caridina indistincta Cal@ ''&M'9'x'

>;+'-F;+-& Page 4 of 13 '+'99''G;+'-+-& >`icrophallid larvae in Australia Deblock and Canaris (1997) Deblock and Pearson ('ZFZ) Deblock and Canaris ('ZZF) Smith ('Z<&) Smith ('Z<&)=J7$and Canaris ('ZZF) Deblock and Canaris (1997) ('ZFZ)=!and Pearson (1991) QueenslandQueensland Deblock and Pearson ('ZFZ) Deblock and Pearson ('ZFZ) = Tasmania != != Queensland Tasmania Smith (1974) = Tasmania Queensland Deblock and Canaris ('ZZF) = Tasmania Queensland Deblock and Pearson ('ZF<) Queensland= Tasmania Queensland Deblock and Canaris ('ZZF) Tasmania Hickman (1955) = Tasmania Tasmania Smith (1974) Queensland Mawson et al. ('Z

bution in Australia. bution in Limosa (Ste@ Arenaria Arenaria Egretta Egretta xQ{=

= Elseyornis xQ{= = Fulica atra xQ{= Temminck xU{= Charadrius mongolus (Stephens) Tachybaptus novaehollandiae Tachybaptus = Tringa hypoleucos Tringa x?p.)

Thalasseus bergii x]!7{= Anhinga melanogaster Blumenbach Q= Geoffroy Thinornis cucullatus xU{= Calidris acuminata Calidris acuminata Thalasseus bergii = = Pallas= x{= (Linnaeus) x[{ x!_{= (Latham) xB{= xQ{= xU{ xQ{= - Chroicocephalus novaehollandiae Chroicocephalus melanops Chickensx?p.)Anas castanea Queensland Shimazu and Pearson (1991) Charadrius mongolus interpres Charadrius rubricollis {= novaehollandiae lapponica Hydromys chrysogaster Hydromys , (Sayce) 6$J

Johnston x6?{ 6$ Metapenaeus bennettae -- Pluvialis dominica Charadrius mongolus -- Pluvialis dominica - chrysogaster Hydromys Caridina nilotica -Coxiella badgerensis australis Austrochiltonia - Charadrius mongolus - anatinus Ornithorhynchus Paragrapsus gaimardii Anas platyrhynchos - Charadrius mongolus Paragrapsus gaimardii x_B`{ - Charadrius leschenaultii - Calidris acuminata - interpres Arenaria Coxiella badgerensis Poliocephalus poliocephalus Cherax dispar Shimazu et Pearson, 1991 Hickman, 1955 Deblock et6'ZF9 Deblock et Canaris, 1997 Deblock et'ZFZ Deblock et 'ZZF Smith, 1974 !'Z<& Deblock et Canaris, 1997 sp. Deblock et'ZFZ Ward, 1901 Ward, Smith, 1974 Deblock et Canaris, 1997 ]'Z9< Deblock et'ZZF (Lebour, 1907) (Lebour, sp. sp. ‰$'Z-; Yamaguti, 1939 Yamaguti, Belopol'$'Z-; Nicoll, 1907

'Z;< 6#7Maritrematinae Nicoll, 1907 and Microphallinae Ward, 1901 and data on their hosts distri Ward, 6#7Maritrematinae Nicoll, 1907 and Microphallinae Microphallus M. minus M. papillornatus M. vaginosus R. insularegii R. insularegii M. rubeum M. ornithorhynchi M. oocysta M. eroliae M. brevisacciferum M. brevisacciferum M. spinosulum M. calvertensis M. paragrapsi A. coxiellae E. incana Maritrema Maritrema M. pearsoni M. minutus Rhyncostophallus Microphallus Atriophallophorus Maritrema Endocotyle Subfamily Microphallinae Ward, 1901 Ward, Subfamily Microphallinae Table 2. Table SpeciesSubfamily Maritrematinae Nicoll, 1907 Second intermediate host J[# 6 Source

>;+'-F;+-& Page 5 of 13 '+'99''G;+'-+-& >`icrophallid larvae in Australia

Table 3. Comparative metrical data for metacercariae of Maritrema brevisacciferum Shimazu et Pearson, 1991 and Microphallus minu- tus ]'Z9<_

Species Maritrema brevisacciferum Maritrema brevisacciferum Microphallus minutus Microphallus minutus Host Caridina indistincta Calman Caridina niloticax6?{ Cherax dispar 6$ Cherax dispar Source Present study Shimazu and Pearson (1991) Present study Shimazu and Pearson (1991) 6 Mean 6 6 Mean 6 Cyst length 'F'M;'- '9{Y` ;&; ly oval, & Posticobia brazieri (Smith) <*M';;x'+'{`?`# (Gastropoda: Tateidae). $9+MFFx-&{Y#` Locality: Moggill Creek, Queensland, Australia. Y F+MZF x<9{ '+M'& x''{ ` 7 Prevalence of emergence: ';'F+ shorter than body. Oral sucker subterminal, muscular, sub@ Second intermediate host: Cherax dispar 6$(De@ ;&M&+&;+M&'x;F&;*=š'-{!

>;+'-F;+-& F'& doi: 10.14411/fp.2015.053 Kudlai et al.: Freshwater microphallid larvae in Australia

Fig. 2. Microphallus minutus Johnston, 1948. A – cercaria from Posticobia brazieri (Smith), ventral view; B – stylet, ventral and lateral view; C – metacercaria from Cherax dispar Reik.

capoda: Parastacidae). Locality: Moggill Creek, Queensland, Australia. Prevalence: 1 of 1. Representative sequences: Cercaria KT355823 (28S rDNA), KT355829 (ITS2), metacercaria KT355822 (28S rDNA), KT355828 (ITS2). Remarks. The metacercapariae from our collection ex- hibit features typical for the Microphallinae Ward, 1901, in particular the absence of a cirrus-sac and the seminal vesicle and pars prostatica lying free in the parenchyma (Deblock 2008). Seven species of Microphallus have been reported in Australia and three of them are known from second in- termediate hosts (Table 2). The present metacercaria is strongly consistent with the morphological description of M. minutus by Shimazu and Pearson (1991) from C. dispar in the shape and structure of the cyst, whereby the outer layer in young metacercariae is very thick. Although no Fig. 3. Microphallidae gen. sp., metacercaria from Posticobia metrical data for the metacercariae are available, we com- brazieri (Smith). pared the encysted metacercariae with a well-developed reproductive system with the metrical data given for adults The species was described based on material from of M. minutus by Shimazu and Pearson (1991) (see Table Hydromys chrysogaster Geoffroy in the Murray Riv- 3). Comparisons revealed that the metrical data of our de- er (South Australia) by Johnston (1948) who suggested scription fall within the range for M. minutus, but our mate- Cherax destructor Clark as potential second intermediate rial exhibits slightly lower limits for the size of the pharynx host of M. minutus. Deblock and Pearson (1969) reported (18 × 15 μm vs 22–24 × 16–17 μm) and the width of the M. minutus from H. chrysogaster in Moggill and Palm- ovary (38–45 μm vs 48–80 μm). Since the thick outer layer wood, Queensland, Australia. Shimazu and Pearson (1991) of the cyst was included in measurements by Shimazu and found metacercariae of this species in C. dispar from the Pearson (1991), the size of the cyst differs from our data in Brisbane River (Queensland), [ distinctly higher upper limits (238–320 μm × 235–297 μm of Johnston (1948). Notably, P. brazieri has been reported vs 310–540 μm × 290–540 μm). widely in eastern Australia (Clark 2009), including from

Folia Parasitologica 2015, 62: 053 Page 7 of 13 doi: 10.14411/fp.2015.053 Kudlai et al.: Freshwater microphallid larvae in Australia

Fig. 4. Phylogenetic relationships among the taxa of the Microphallidae resulting from Bayesian inference (BI) and Maximum likeli- hood (ML) analyses based on partial 28S rDNA data. Nodal support associated with the branches are listed as BI posterior probability/ ML bootstrap support; support values lower than 0.90 (BI) and 70 (ML) are not shown. The scale-bar indicates the expected number of substitutions per site. Sequences obtained in the present study indicated by bold font. Abbreviations: C – cercaria; M – metacercaria. the type locality of M. minutus, Tailem Bend on the Murray (Decapoda, Atyidae). River in South Australia; presumably it is also involved in Locality: Fairnie Brook and Churchbank Weir, Queensland, transmission there. Australia. The sequences of cercarial isolates from the snail Prevalence: C. indistincta 8% (Churchbank Weir). P. brazieri and metacercarial isolates from C. dispar were Representative sequences: KT355820, KT355821 identical for each of the 28S and ITS2 rDNA regions. We, (28S rDNA), KT355826, KT355827 (ITS2). therefore, consider that they belong to the same species. Y [ Remarks. Further study on the morphology of this M. minutus. species is required. This species may well be close- # [ ` Microphallus Microphallidae gen. sp. Fig. 3 xΣ#Τ{  x;+';{ Potamopyrgus antipodarum from New Zealand, which also develops Description of metacercaria (preliminary description directly to metacercariae in the tissues of the snail. based on 24 live encysted specimens from P. brazieri from Abbreviation of microphallid life cycles by formation of Fairnie Brook – 19 and C. indistincta from Churchbank [x Weir – 5): Metacercaria in subspherical cyst, 115–129 × $#!$;++*{`# 105–123 (123 × 114). Cyst wall consisting of two hyaline to such development in the present case, our recording of layers. Entire tegument covered with dense spines. Oral genetically and morphologically identical metacercariae sucker subterminal; ventral sucker large, muscular. Repro- from atyid shrimps shows that the present form has two ductive system well-developed. Testes lateral, oval, sym- potential pathways of transmission. We are unaware of metrical. Structure resembling phallus large, almost equal previous reports of such a dichotomy. to ventral sucker in size, sinistral to ventral sucker. Vitel- larium in two symmetrical clusters of few large follicles, Phylogenetic analysis post-testicular in hindbody. Excretory vesicle Y-shaped. The specimens of M. brevisacciferum, M. minutus and Second intermediate host: Posticobia brazieri Microphallidae gen. sp.`[j (Smith) (Gastropoda, Tateidae), Caridina indistincta Calman variability for either the 28S or ITS2 rDNA regions. The alignment used in the phylogenetic analysis included the

Folia Parasitologica 2015, 62: 053 Page 8 of 13 '+'99''G;+'-+-& >`icrophallid larvae in Australia

`@;9{Y[ @# Microphallus. Sequences for M. minutus generated during 7#x;++&Y$ this study clustered with strong support together with oth@ ;++& ` ;+'9{Y ` er species of Microphallus. The second strongly support@ `@[ ';Maritrema with which the j7Y$x;++&{ new sequences for M. brevisacciferum formed a monophy@ x;++&{ letic lineage associated with strong support with sequences Life cycles of microphallids take place in brackish, for Maritrema spp. from a P. antipodarum marine or fresh waters, in decreasing order of frequency and a bird Anas platyrhynchos (Linnaeus), described from xJ7$;++<{Y#5$`[@ R`Kx`;+'9{ diate hosts are littoral gastropods of the genera Hydrobia Our phylogenetic analyses clearly indicated that Mi@ Hartmann and Littorina> x‰$'ZF& crophallidae gen. sp. forms a third distinct clade, together J7$'Z<+ $#'ZZZ # `[Microphallus fusiformis6@ ;+++ $#;+';{&'@ 'ZF& # 7 Maritrema (see lid species are currently known to develop in fresh water. below). Obtaining rDNA sequences from additional rep@ They are mainly reported from second intermediate hosts resentatives of the Microphallidae would be essential for 7?7xY7 [ 4). In our analyses, three species of Maritrema that develop in freshwater invertebrates (Maritrema neomiY$'ZZ< DISCUSSION M. brevisacciferum and Maritrema poulini), clustered in Using combined morphological and molecular charac@ a strongly supported subclade nested among species devel@ teristics, we were able to establish links between cercar@ #7= iae and metacercariae of Maritrema brevisacciferum and species in the subclade, Maritrema deblocki Presswell, Microphallus minutus for which complete life cycles were @#;+'9remains unknown. #$`=`#[ Posticobia brazieri has traditionally been included in their cercariae. M. minutus@` 7!x$;++Z{`#` cercariae developing in the tateid snail Posticobia brazieri have here recognised it as belonging to the Tateidae in line [Cherax `QR@ dispar. Adults develop in mammals, water rats (Hydromys 7€$x;+'&{Y@ chysogaster= { x? { [P. brazieri, but it is clear that the scope of Cercariae of M. brevisacciferum also develop in P. bra- the family is thought to incorporate it. The report of three zieri and metacercariae develop in atyid shrimps, Carid- species of the Microphallidae from P. brazieri here adds ina indistincta= [# $` to the remarkable list of trematodes known to infect this _j$[`[ [ > hosts being dependent on trophic habits of predators for `'Z';x crustacean or, occasionally, molluscan second intermediate MR77;++9{@ xJ7$;++<{= togonimidae Ward, 1917, one species of the Haploporidae [>7 Nicoll, 1914, two species of the Lepocreadiidae Odhner, [ 'Z+-$'Z;- congeners. species of the Transversotrematidae Wittenberg, 1944 (see The new sequences of the metacercariae of Microphal@ 77'Z<-'Z formis7[j the only rissooid gastropod that we have seen in fresh wa@ M. fusiformis is unclear. The annotation in GenBank (Tk@ @[

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(Savigny) Glossiphonia complanata Hobbs,

Say x6?{ Chicken (Linnaeus) Schmitt, (Say) 6$ Moore, (De Haan), Dana (Leidy) Helobdella stagnalis Paracorophium excavatum Paracorophium Palaemonetes argentinus angulatus Austridotea annectens (Thomson), Gammarus aequicauda haemobaphes Dikerogammarus Gammarus pulex Galemys pyrenaicus Neocaridina denticulata Rostratula benhalensis Procambarus clarkii Procambarus antipodarum Potamopyrgus nipponense 6$J x @_ #{ Gammarus balcanicus Cambarus propinquus punctata naeus), Gammarus balcanicus Asellus communis Cambarellus puer Cambarellus

Littoridina (Gray) Gredler (Dupuy) Pilsbry et Walker Pilsbry et Walker Littoridina parchappei Palaemonetes argentinus x>{ (Gaillard), (Say) , sp. (d’Orbigny) Heleobia conexa parchappei -Amnicola pilsbryi Asellus communis Gammarus balcanicus Oncomelania nosophora - Cherax dispar Potamopyrgus antipodarum Potamopyrgus - nipponense Macrobrachium - Caridina nilotica ---- Erpobdella testacea Gammarus pulex - Obesogammarus crassus Lontra provocax Mercuria confusa Mercuria - Neocaridina denticulata - Aegla neuquensis Amnicola limosa reyniesii - Cambarus propinquus Heleobia conexa - Pseudothelphusa chilensis - - asperulus Macrobrachium Terrestribythinella baidashnikovi Terrestribythinella Anistratenko Sitnikova, Starobogatov et Pseudamnicola Amnicola pilsbryi Amnicola peracuta - Dina parva - Amnicola limosa - -australiense Macrobrachium 7'Z*F x!@'ZF;{ Y@J#'ZF+ xU#_'Z&;{ Shimazu et Pearson, 1991 (Yamaguti et Nisimura, 1944) (Yamaguti Tsai, 1955 Tsai, Y@J#'ZF; $5$#'Z<& B'Z-& 6j>;+'& ]'Z9< RJ€'Z<- J7$7'ZF- x€';+'; #Q'ZZ; `@# Y$'ZZ< Gracenea, Montoliu et Deblock, 1993 - Cambaroides japonicus - Cambaroides xΣ#Τ{ J7$'ZF< Stenko, 1973 Species of the Microphallidae with freshwater life cycles. sp. poulini ‰$x'ZF&{=;MJ7$x'ZF<{=&M!x'ZZ'{=9MY@J#x'ZF;{=-M Microphallus szidati Microphallus M. tauricus Levinseniella amnicolae Quasimaritrema caridinae Quasimaritrema Microphallus minutus Microphallus ;+'9 Table 4. Table SpeciesPseudolevinseniella cheni > Second intermediate host J[# 6 Source 1 – Maritrema brevisacciferum Maritrema erpobdellicola Maritrema galloprovinciale Maritrema huillini Maritrema inusitata Maritrema Maritrema feliui Maritrema Maritrema mapensis Maritrema Maritrema patagonica Maritrema Maritrema opacus Microphallus Maritrema pyrenaica pyrenaica Maritrema Maritrema medius Quasimaritremopsis Levinseniella cruzi Megalophalloides apanhorayi fonti Microphallus Levinseniella minuta Microphallus minus Microphallus 10 – $5$#x'Z<&{=''MBx'Z-&{=';M6jx;+'&{='&M`x;+'9{='9MCasanova et 10 – $5$#x'Z<&{=''MBx'Z-&{=';M6jx;+'&{='&M`x;+'9{='9MCasanova 7$x'ZFZ{=;+M]x'Z9<{=;'M6x'Z9*{=;;M#Bx'Z*'{=;&Mx;+';{=;9MM !@x'ZF;{=&+MQ$x'Z<&{=x?{M?7 P. anenteron anenteron P. Maritrema neomi Maritrema parainusitata Maritrema obstipum Maritrema Sogandatrema progeneticus progeneticus Sogandatrema Levinseniella ophidea Microphallus Microphallus

>;+'-F;+-& Page 10 of 13 '+'99''G;+'-+-& >`icrophallid larvae in Australia which often occurs in huge densities, plays a pivotal role three species of Hydrobia from salt marshes and estuaries in a wide variety of the trematode cycles in the region. >=$79+ !`[7 from 14 families. Microphallids occupy a central position `x;+';{$`;+ `';!Hydrobia are current@ species of trematodes that infect the New Zealand tateid, ly known as an intermediate host for about 19 species of P. antipodarum. The parallels with the fauna of P. bra- _xJ7$'Z*<'Z<+>` zieri are striking. Both harbour Aporocotylidae (both two 'ZZZ 7;++*;+''{ species), Cryptogonimidae, Lepocreadiidae, Microphalli@ dae, Opecoelidae, Pronocephaloidea, and (possibly) Hap@ Acknowledgements. Y$6[@~~ 7BJ`$#`@ loporidae and Psilostomidae Looss, 1900. The only fami@ Y`7B ly level distinctions between the two are the strigeid and M!>5Q>`@ Q@$RP. antipodarum and the QR xU'@&'@__@+'@U@+;@++9{ and the presence of hemiuroid, renicolid and transversotrematids !>x5B-+-G';G '';{ in P. brazieri>#@ 76!Y !J7$x'Z*<{ and SCC.

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Ǎ@ǂǏDžǂǓNJ€~Ǎ@ǖǔǕǂǏ!ǍǏǂǒdždžǃ_;+''67@ ǓNJǃǃY'Z<<7Y`` somal DNA sequence characterization of Maritrema cf. eroliae `[`#7] Yamaguti, 1939 (Digenea: Microphallidae) and its life cycle. J. R;;;*M9& Z*'+F*M'+*9 ǓNJǃǃYǏDždžǓǔǐǏ 6DžǍǂǓDž6JǓǂǚ6'ZZ< ǏDždžǓǔǐǏ 6ǂǓnjdžǓ!'ZZ< JR@7 @ @ ?`JxJ{ # x J{ ] ;< 7xY!;{7JR!@ 1791–1795. 9'<*MZ9 JǂǓǓNJǃǂJYǂǃǐǂDžǂ QJǐǂǍǍǐ6ǐǔǂDžǂJ;+';5_@ ǂǓǕǐǍNJ NJǃǔǐǏJ;++*!J@ Y;` nea (Platyhelminthes) from lagoons of the northern coast of the R_Z**; western Mediterranean. J. Nat. Hist. 41: 1553–1570. JdžǃǍǐDŽnj ! 'Z*<J7 j@ džǍǐǑǐǍ‰ǔnjǂǚǂ __ 'Z-; > _ž sites des mollusques du genre Hydrobia Hartman des côtes de !$57xB{#YUF‰# >-&-**M-Z& $R$!!!6_$#F'ZM*-Fx6{ JdžǃǍǐDŽnj!'Z<+# # džǍǐǑǐǍ‰ǔnjǂǚǂ__'ZF&>_ž!$@ mollusques Hydrobiax7{>@ 57 xB{ # Y U ;' ‰# ;;'M'+- $R$!!!6_$#;F+M-+;x6{ JdžǃǍǐDŽnj!;++<>_€'Z+'6 ǐǓDžǂǍǐ _J >džǓǓdžNJǓǂ !_ ]džǏǔdžǏ Y ǂǓDžǂǍ _ J 7]xB{Y ;+''YHydrobia ulvae (Gastropoda: Proso@ U&€R_ branchia) in a eutrophic temperate estuary. J. Mar. Biol. Ass. UK Q9-'M9Z; Z'Z'&MZ;' JdžǃǍǐDŽnj!ǂǏǂǓNJǔ'ZZF_YÃQU@ ǐǘǍdžǔ]ǐǑdž_YNJǖ€MQNJǖÃ!_DŽ_ǂǏǖǔJ1993: II. Quatre MaritremaB=R?@ Nuclear and mitochondrial genetic markers highly conserved 9&-*M&F' between Chinese and Philippine Schistosoma japonicum. Acta JdžǃǍǐDŽnj!ǂǏǂǓNJǔ 'ZZ*7R _@ Y--;'*M;;Z xY{ ÃQà B R? Y@ ǂǏǏǐǏQ6 'Z*<_ Ce- Rhyncostophallus n. g. Syst. Parasitol. 37: rithium moniliferum 6 13–19. 6![Mar- Y#'Z;+xY{ÃUJjj@ itrema pyrenaica (Digenea: Microphallidae) in Spain (Western pèces d’Australie dont Pseudospelotrema anenteron n. sp. Ann. {>9-;-'M;-; 9&9-*M9F- ǂǗdžǏǚBǕLjdžǔ>1971: Life history studies of Microphallus JdžǃǍǐDŽnj!džǂǓǔǐǏ]'ZFZ7R _@ opacusxY_{]-*';'-M';;' Y#'Z;+xY{ÃUMJj ljNJǏLj Q ǃǂDzdžǛ R 'Z*FJMegalophalloides MicrophallusR?#? apanhorayi gen. et sp. n. (Trematoda, Microphallidae) from Hum. Comp. 44: 391–414. `7]K-9'9&

>;+'-F;+-& Page 11 of 13 '+'99''G;+'-+-& >`icrophallid larvae in Australia

telidae) from freshwater environments of northwestern Patago@ _ǂǓǕǐǓdžǍǍNJ!6'ZQdžLJǐǓǕUǏNJǔNJǎǐǗǂ_ǐǓDž@ RǐǍǂǏ_]ǓNJǃǃY;++9YParacardicoloides NJNjnj€ ǂǔDŽǖdžǍ;+'+R`@ yamagutii_'Z*9xJ!{>@ ?@$@ -'&;+M&;F _Q&+!-Z&+*M&;' RJǘǚdžǓ ǍǂǔDŽǐ@ǐǔǕǂ ǐǖǍNJǏ 6 >ǂǍǕǼǏnjǐǗǤ ǂǍǍ Y 'ZZZ B @ 7 j ;+'9 > Austrolittorina spp. #-+ZR!!9'Z-MZ< (Gastropoda: Littorinidae) in New Zealand: morphological and džDŽljNJǏLjdžǓ6>;+';>#[$ !Y#'9_#@ >F;++& B7 M M# B@ ǖǍnjNJǏǂQUdžǍNjǂnjǐǗǂ~ǖU'Z<&QMaritrema 7B#GG7$G parainusitataxY_{ž@ `G[G';G;+'; '*;*;M;**x6{ 6ǂǖǔDŽlj 6 1947: Some observations on the host relationships of ǖǓǂǏDžNJǏǂJ'Z*<[Maritrema inusitata Le@ Microphallus opacus x€ 'YdžǔǍdžǏnjǐ_UǂǏJdžǓ_ǂǓnjǚǓdžǔJQ R$#$#';&M';-x6{ JǂǓǍNJǏLjǷljǏǂ!QǂǓLjdžǕQNJǖQ!ǖDŽljǂǓDž_ QdžǐǏǐǗUYǔNJǎǃǂǍǚǖnj'ZF&`@ ǖdžǍǔdžǏǃdžDŽnj];+';_&;B[@ tode of Maritrema inusitata@$ logenetic inference and model choice across a large model space. $žU$Q$M#''9-M'9Z !F'-&ZM-9; x6{ 6ǂǖǒǖdž>ǍǐǓdžǔU6ǓǖLjǏNJRQ;+'&Maritrema pata- QNJǕǕǍdžǘǐǐDžJY]ǖǓNJǏNJ@ ǂǍǍdžǕǕNJ_džǓǏNJǐǖB;+++ gonica n. sp. (Digenea: Microphallidae) cultured from metacer@ The interrelationships of Proseriata (Platyhelminthes: Seriata) cariae from freshwater anomuran, Aegla spp. (Decapoda: Aegli@ `_B# {<+'ZFM;+; 'F99ZM9FF !ǂǎǃǓǐǐnj]6ǖǔǔdžǍǍJ€;++'_Q7@ QǐǕǛ ]_ ǐǓnjǖǎ 'Z<& ! tory Manual. Cold Spring Harbor Laboratory Press, Cold Spring Sogandaritrema progeneticus (Digenea: Microphallidae). J. 7;'++ FZZ'

>;+'-F;+-& ';'& '+'99''G;+'-+-& >`icrophallid larvae in Australia

!ǏǚDždžǓ!JYnjǂDŽljUU;++'7 YnjǂDŽljUUQNJǕǕǍdžǘǐǐDžJY]ǍǔǐǏJNJǏǔdžǍǍǂ]_ \$xJ !ǘNJDždžǓǔnjNJK;++&__@ {]<*'9&&M'99+ crophalloidea Ward, 1901 (Trematoda: Digenea). Syst. Parasitol. !ǐLjǂǏDžǂǓdžǔ@džǓǏǂǍ>'ZF;Microphallus progeneticus, a new -F'M'- apharyngeate progenetic trematode (Microphallidae) from the YnjǂDŽljUUǂǘǍǐǘǔnjNJ]_ǂǓNJǂǖǙ];+++@ ` [ Cambarellus puer, in Louisiana. Tulane Stud. ysis of the suborder Plagiorchiata (Platyhelminthes, Digenea) KZ&'ZM&;; 7JRj]&+<&MZ& !ǕdžǏnjǐ61973: [Microphallus tauricus sp. n. (Microphallidae €ǂǕǔǐǏ 6 'Z<9 Y Tetracer- Y#'Z;+{`ž asta blepta gen. et sp. nov. and Stegodexamene callista sp. nov. *-'&M-'*x6{ xYQ{@[Anguilla !ǕǖǏnjǂǓDž € 'Z-<Y@Lev- reinhardtii!]K&;'**M;+9 inseniella minuta (Trematoda: Microphallidae). J. Parasitol. 44: €NJǍnjdžYǂǂǔdž_džǓǔljǍdžǓ6QNJǖ@_NJǔǐLJǐǏ@ ;;-M;;Z DždžǓ€;+'&JR@ YNJǎǐǏ@JǂǗNJDž]'ZF;M Maritrema Q7R x@ Nicoll (Trematoda, Digenea, Microphallidae) parasite de 6{_B#FF*'-M*&F 'R Erpobdella testaceax!#{!K> €ǖ1951: Progenesis of Microphallus minus Ouchi in fresh wa@ <*--ZM-F- $R'Z'Z9M;+Z Ynj ǂDŽlj UU 'ZZ<Maritrema neomi n. sp. (Digenea: Microphalli@ dae) from water shrews (Neomys{]<9<9FM<9Z

6# ;' _ ;+'- Z ] ;+'- 7 'F !7 ;+'-

Cite this article as: !77Y;+'-__@ xYJ{[Maritrema brevisacciferum and Microphallus minutus>F;+-&

>;+'-F;+-& Page 13 of 13