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Wallaceaphytis: an unusual new genus of parasitoid wasp (Hymenoptera: Aphelinidae) from Borneo


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Wallaceaphytis Polaszek and Fusu gen. nov. (type species Wallaceaphytis kikiae Ayshford and Polaszek sp. nov.) is described from Danum Valley, Sabah, in Malaysian Borneo. Although known from just a single female individual, the genus is extremely unusual morphologically, being the only member of the large subfamily Aphelininae with four-segmented tarsi. The form of the fore wings and head are also unique in the subfamily, and its status as a new genus is confirmed by analysis of nuclear ribosomal DNA. DNA sequence analysis was undertaken by comparison with more than 60 aphelinid sequences from GenBank. The sequence for the standard DNA barcode region (cytochrome oxidase c subunit I; COI) is provided. The new genus is named in honour of Alfred Russel Wallace, co-discoverer of the theory of evolution by natural selection. The new genus and species are published on the exact date of the centenary of his death.
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Wallaceaphytis: an unusual new genus
of parasitoid wasp (Hymenoptera:
Aphelinidae) from Borneo
Andrew Polaszeka, Thomas Ayshforda, Bakhtiar Effendi Yahyab &
Lucian Fusuc
a Department of Life Sciences, Natural History Museum, London,
b Institute for Tropical Biology and Conservation, Universiti
Malaysia Sabah, Kota Kinabalu, Malaysia
c Faculty of Biology, Al. I. Cuza University, Iasi, Romania
Published online: 07 Nov 2013.
To cite this article: Andrew Polaszek, Thomas Ayshford, Bakhtiar Effendi Yahya & Lucian Fusu
(2014) Wallaceaphytis: an unusual new genus of parasitoid wasp (Hymenoptera: Aphelinidae) from
Borneo, Journal of Natural History, 48:19-20, 1111-1123, DOI: 10.1080/00222933.2013.852264
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Wallaceaphytis: an unusual new genus of parasitoid wasp (Hymenoptera:
Aphelinidae) from Borneo
Andrew Polaszek
*, Thomas Ayshford
, Bakhtiar Effendi Yahya
and Lucian Fusu
Department of Life Sciences, Natural History Museum, London, UK;
Institute for Tropical
Biology and Conservation, Universiti Malaysia Sabah, Kota Kinabalu, Malaysia;
Faculty of
Biology, Al. I. Cuza University, Iasi, Romania
(Received 2 October 2013; accepted 2 October 2013; first published online 7 November 2013)
Wallaceaphytis Polaszek and Fusu gen. nov. (type species Wallaceaphytis kikiae
Ayshford and Polaszek sp. nov.) is described from Danum Valley, Sabah, in
Malaysian Borneo. Although known from just a single female individual, the
genus is extremely unusual morphologically, being the only member of the large
subfamily Aphelininae with four-segmented tarsi. The form of the fore wings and
head are also unique in the subfamily, and its status as a new genus is confirmed
by analysis of nuclear ribosomal DNA. DNA sequence analysis was undertaken
by comparison with more than 60 aphelinid sequences from GenBank. The
sequence for the standard DNA barcode region (cytochrome oxidase c subunit
I; COI) is provided. The new genus is named in honour of Alfred Russel Wallace,
co-discoverer of the theory of evolution by natural selection. The new genus and
species are published on the exact date of the centenary of his death.
Keywords: Aphelininae; Chalcidoidea; chalcids; phylogeny; Sabah; Malaysia;
Alfred Russel Wallace; DNA barcode; non-destructive DNA extraction
Within the large and megadiverse parasitoid superfamily Chalcidoidea (chalcid
wasps), Aphelinidae is one of the smaller families, containing 1300 species belonging
to 36 genera (Noyes 2013). Species of Aphelinidae are mostly primary parasitoids or
hyperparasitoids of Hemiptera, mainly Sternorrhyncha (Aleyrodidae, Aphididae,
Coccidae, Diaspididae & Pseudococcidae, among others), although several genera
are known to include species that are parasitoids of insect eggs (Polaszek 1991).
The large subfamily Aphelininae was recently the subject of a major phylogenetic
analysis based on morphological characters, which resulted in the description of four
new genera (Kim & Heraty 2012). This analysis includes a key to all of the currently
recognized 16 genera within the subfamily. The new genus described below differs
radically in several characters from any of the currently valid genera of Aphelininae,
and these differences are discussed in detail below.
*Corresponding author. Email:
Journal of Natural History, 2014
Vol. 48, Nos. 1920, 11111123,
© 2013 Crown Copyright published by Taylor and Francis
This is an Open Access article. Non-commercial re-use, distribution, and reproduction in any medium, provided the
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Material and methods
In September and October 2012 a major multidisciplinary joint expedition from the
Natural History Museum, London, UK, and Universiti Malaysia Sabah, Kota
Kinabalu, Malaysia, undertook extensive sampling and surveying of arthropods
and other invertebrates in the Danum Valley and Maliau Basin Conservation
Areas. A wide range of collecting techniques was employed, including the use of
Winkler bags for soil samples, Malaise traps and yellow pan traps for day-flying
insects, and a specially modified sweep net for very small insects found in under-
growth and foliage. The Noyes-net(Noyes 1982) is a heavy-duty, long-handled
sweep net with 4-mm wire mesh screening the collecting bag, ensuring that only
specimens with a maximum length of 4 mm or thereabouts are collected. Sweeping
the undergrowth and foliage for several minutesresultsinanaccumulation of usually
several hundred microarthropods in the collecting bag, as well as associated debris,
seeds etc. These can either be collected directly into a container of 80100% ethanol
for sorting under a microscope later, or the emergent insects can be aspirated indivi-
dually. In the latter case, less sorting is needed subsequently, but a large proportion of
the catch can be lost or overlooked. The latter technique was used in the present case,
with the sample sorted back in London several weeks later. Immediate recognition by
the second author (TA) that a particular specimen was clearly something extremely
unusual, prompted a special study of this individual, which was then subjected to the
non-destructive DNA extraction protocol described below.
DNA extraction and slide-mounting
Genomic DNA was extracted using the DNeasy
blood and tissue kit (Qiagen,
Hilden, Germany) from the whole specimen, using a non-destructive method slightly
modified from the manufacturersprotocol(Noyes2010). The specimen was removed
from ethanol, dried briefly on absorbent paper to remove any visible traces of liquid,
and immersed in ATL lysis buffer containing proteinase K in a 1.5 ml microtube
). Specimen lysis was achieved by overnight incubation at 55°C (or for
at least 8 hours) after which all internal tissues have been digested, while the exoske-
leton remains intact in the enzymebuffer mix (throughout this time vortexing should
be avoided to reduce the risk of damaging the specimen). The lysis buffer containing
DNA was transferred by pipetting to a new 1.5 ml microtube, and processed as
described in the kit, except that the final DNA elution was into 100 µl. The extracted
specimen was immediately washed by pipetting 1 ml distilled water into the micro-
tube, changing the water after 30 min, and finally transferring the specimen into 80%
ethanol. If the specimen is not thoroughly washed, crystals can form upon its surface
on contact with ethanol. In the unlikely case of crystal formation, these can be
dissolved by placing the specimen for a few minutes in warm distilled water.
Specimens extracted with this method can be directly mounted without a previous
alkaline treatment (e.g. maceration in 10% KOH), or critical-point dried or chemi-
cally dried using hexamethyldisilazane, but air drying is likely to result in the specimen
collapsing. For slide-mounting, the specimen can be dehydrated through graded
alcohols up to 100% and permanently slide-mounted in Canada balsam after clearing
with clove oil. In the present case, the specimen was dissected and mounted following
the protocol described by Noyes (1982), from the 100% ethanol/clove oil stage
1112 A. Polaszek et al.
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onward, with the modification that dissection takes place in Canada balsam to reduce
specimen movement and the consequent risk of losing the dissected parts.
Polymerase chain reaction (PCR) and sequencing
The standard barcode region (Hebert et al. 2003) and the 28S rDNA D2 and D3
expansion regions were amplified by PCR using the LCO1490/HCO2198 primer pair of
Folmer et al. (1994)andD23F(5-GAG AGT TCA AGA GTA CGT G-3;Park&
Foighil 2000)/28Sb (5-TCGGAAGGAACCAGCTACTA-3, aca D3B; Nunn et al.
1996), respectively. We performed standard 25-μl PCRs containing 2.5 μl of 10× PCR
buffer, 0.75 μlof50m
,0.2μl dNTPs solution (25 mMeach), 1.25 μlofeach
primer (10 μM), 0.3 μlTaq polymerase (5u/μl Biotaq, Bioline), 6 μl DNA extract, and PCR
grade water to final volume. PCR conditions for cytochrome oxidase c subunit I (COI)
were 94°C for 2 min, followed by 40 repeated cycles of 94°C for 30 s, 42°C for 50 s and 72°
C for 35 s, a final extension at 72°C for 10 min and incubation at 10°C. The same
conditions were used for the amplification of 28S rDNA except annealing for 30 s at 55°
C. The PCR products were visualized on a 1% agarose gel.
Both DNA strands were sequenced at the Natural History Museum Life Sciences
DNA Sequencing Facility using the same primers used for the PCR. Sequences were
edited using Pregap4 v1.5 and Gap v4.10 in Staden Package (Bonfield et al. 1995) and
sequence verification was conducted by comparing forward and reverse sequences.
All sequences are deposited on GenBank (Accession numbers BankIt1665361 Seq1
KF718961 [28S ribosomal]; BankIt1665361 Seq1 KF718962 [CO1]).
Additional sequences for the phylogenetic analysis obtained mostly by J.-W. Kim
and J. Heraty (unpublished data) were retrieved from GenBank. Sequences were first
aligned using MEGA 5.05 (Tamura et al. 2011) and the ClustalW algorithm for the
ML analysis, or were manually aligned using the secondary structure models follow-
ing Gillespie et al. (2005) and the alignment of Munro et al. (2011) for the Bayesian
analysis. Phylogenies were estimated using maximum likelihood in MEGA 5.05 and
Bayesian methods in MrBayes version 3.2 (Ronquist & Huelsenbeck 2003). Analyses
were run using a GTR + G + I model of nucleotide substitution as this was
determined as the most appropriate model with MEGA 5.05. In MrBayes the analysis
was run for 10,000,000 Markov Chain Monte Carlo generations, with trees and lnLs
sampled every 100 generations. Likelihood stationarity occurred after 15,000 genera-
tions that were discarded as burn in.
Sequence analysis
The aligned sequences of the 28S rDNA D2 and D3 expansion region produced a
sequence matrix 1121 base pairs long when automatically aligned using the ClustalW
algorithm or 1074 base pairs long in the case of the secondary structure alignment.
Maximum likelihood analysis resulted in one best-scoring tree with Wallaceaphytis
placed as sister group to a clade formed by all Centrodora species used in the analysis
(Figure 1), but this relation is only weakly supported (54% bootstrap support). In the
bootstrap consensus tree Wallaceaphytis,Centrodora,Aphytis, and Aphelinus are all
part of an unresolved polytomy (tree not shown). In the Bayesian analysis, the exact
position of Wallaceaphytis was not resolved either, as it was placed in a polytomy
with Marietta,Aphytis and Aphelinus (Figure 2).
Journal of Natural History 1113
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Figure 1. Maximum likelihood tree based on 28S-D2 and D3 sequences. Bootstrap values
based on 1000 replications are shown for nodes with more than 50% bootstrap support.
1114 A. Polaszek et al.
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Figure 2. Majority rule consensus Bayesian tree based on 28S-D2 and D3 sequences. Posterior
probability values are indicated in bold above nodes.
Journal of Natural History 1115
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Wallaceaphytis Polaszek and Fusu gen. nov.
Type species Wallaceaphytis kikiae Ayshford and Polaszek sp. nov.
(Figures 311)
Description/generic diagnosis
Morphology. Antenna with three segments (Figure 5), scape, pedicel and a single
flagellar segment. Anellus present, narrower on its internal side. Scape narrow,
length 5× maximum width. Maximum width of pedicel 1.6× maximum width of
scape. Flagellum length 2.9× maximum width, and 1.4× scape. Head strongly trans-
verse (Figures 4 and 8) 2.9× as wide as long in dorsal view (unmounted specimen
Figure 8); 1.3× as wide as maximum width of mesosoma in dorsal view (Figure 8).
Mandible very small, with two teeth and a small truncation; mandibular glands
Figure 3. Wallaceaphytis kikiae holotype female; mesosoma and metasoma.
1116 A. Polaszek et al.
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elongate, parallel-sided. Maxillary palp two-segmented, labial palp one-segmented.
Lateral ocellus separated from eye margin by slightly more than the maximum width
of ocellus. Pronotum centrally membranous, each side with a robust seta at the
lateral edge, a fine seta adjacent to it, and two fine setae further towards the
centre. Mid-lobe of mesoscutum with two setae laterally. Each side lobe of mesoscu-
tum with two setae; tegula with a robust seta; axilla without setae; scutellum trans-
verse, with two pairs of setae. Propodeum elongate centrally, projecting posteriorly,
with a central process, and without crenulae (Figure 3). Propodeal spiracle
without anterior groove. Mesofurca of typical Aphytini form (Figure 7; see Heraty
et al. 1997). All tarsi four-segmented. Fore wing 3.8× as long as maximum width of
disc (excluding marginal fringe); submarginal vein with a single seta; stigmal vein
well-developed (Figure 6). Fore wing without setae below marginal vein, remainder
of wing very sparsely setose. Anterior gastral sterna without projections (see Woolley
1988, p.469). T1T6 of gaster each with a pair of setae, T7 (syntergum) with two pairs
of setae, and in the form of a single sclerite, undivided and without epiproct.
Figure 4. Wallaceaphytis kikiae holotype female; head: dorsal (above), ventral (below).
Journal of Natural History 1117
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Figure 5. Wallaceaphytis kikiae holotype female; antennae.
Figure 6. Wallaceaphytis kikiae holotype female; fore wing.
1118 A. Polaszek et al.
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Wallaceaphytis presents a combination of characters that is so far unique among the
family Aphelinidae. Within the family, Wallaceaphytis is clearly a member of the sub-
family Aphelininae, as shown above based on DNA sequence data, and as follows based
on morphological data. The reduced number of antennal segments (three in the present
case), elongate and parallel-sided mandibular glands, medially membranous pronotum,
propodeal spiracles without anterior grooves, sterna without anterior apodemes,
Aphytis-like mesofurca and presence of a syntergum, exclude all other subfamilies.
Wallaceaphytis superficially resembles Ablerus, especially the unusual fore wing, but
can be easily excluded from that genus and from the subfamily Azotinae (family
Azotidae) by the above combination of characters. Eretmocerus has been included in
Aphelininae by some authors, and also has four-segmented tarsi, while the antennae are
five-segmented in females and three-segmented in males. However, in virtually all other
respects the two genera are very distinct, and Eretmocerus appears to be only distantly
related to Aphelininae. Marlatiella is the only known genus that also has three-segmen-
ted female antennae, but has five-segmented tarsi and very different wing characters.
Molecular DNA analysis supports the status of Wallaceaphytis as a distinct genus
with unresolved affiliations or allied more closely with Centrodora, but this relation-
ship is weakly supported. Despite the Bayesian analysis placing Wallaceaphytis in a
polytomy with Marietta,Aphytis and Aphelinus, morphology, and the maximum
likelihood analysis, suggest a closer relationship with Centrodora. In the key to genera
of Aphelininae by Kim and Heraty (2012)Wallaceaphytis keys immediately with
Eretmocerus because of the four-segmented tarsi, but is easily separated from this
genus by the three-segmented female antenna, and long marginal vein. Although
Wallaceaphytis is superficially similar to Ablerus, the molecular analysis (as with the
comparison of morphology discussed above) clearly shows that this genus belongs to
Aphelininae and not to Azotinae.
Figure 7. Wallaceaphytis kikiae holotype female; mesofurca.
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Figure 9. Wallaceaphytis kikiae holotype female; habitus.
Figure 8. Wallaceaphytis kikiae holotype female; habitus.
1120 A. Polaszek et al.
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Wallaceaphytis kikiae Ayshford and Polaszek sp. nov.
In addition to the genus-level characters above, the following characters are likely to
be of species-level significance if additional species of Wallaceaphytis are discovered:
Colour. Head dorsally orange-yellow, brown on lower half of occiput (Figure 8).
Scape dark brown, pedicel and flagellum orange-yellow, flagellum darker basally.
Mesosoma and metasoma brown-black (Figures 9 and 10), a pale intersegmental area
behind propodeum laterally. Fore wing infuscate from base to slightly beyond stigma
vein (Figures 6 and 9). Legs brown, femora and tibiae paler at their bases; distal tarsal
segments darker than proximal segments (Figure 3).
Sculpture. Frons and face with transverse/reticulate sculpture. Mesoscutum
with reticulate sculpture in the form of large irregular cells, Fore wing
marginal vein with four robust setae and a smaller one at the junction with the
submarginal vein.
Additional characters
Ovipositor projecting beyond metasoma; 2.3× mid tibia. Second valvifers 3.3 × third
Figure 10. Wallaceaphytis kikiae holotype female; habitus.
Journal of Natural History 1121
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Holotype female. MALAYSIA: Borneo, Sabah, Danum Valley Field Study Centre,
Beach. 5°01' N, 117°48.75' E. 14 September 2012 screen-sweep (A. Polaszek).
Holotype dissected and slide-mounted in Canada balsam, deposited permanently in
Institute for Tropical Biology and Conservation, Universiti Malaysia Sabah, Kota
Kinabalu, MALAYSIA.
The genus name Wallaceaphytis is derived from the family name of Alfred Russel
Wallace, and the generic name Aphytis, to which it is related. Other genera within
the subfamily Aphelininae, to which Wallaceaphytis belongs, include Neophytis,
Paraphytis and Punkaphytis. The genus is described in honour of Wallace,
co-discoverer with Charles Darwin of the theory of evolution by natural selection.
Wallaces collections and observations on the fauna and flora during his extensive
travels in South East Asia, including the islandofBorneo,ledtohisformulation
of the theory. The genus and species are described on the exact date of the
centenary of his death.
The specific epithet kikiae is based on, and in honour of, Kiki, the second
authors mother, Mrs Christian Duke.
We thank Mohammad Hayat, John Noyes and Jim Woolley for improving earlier versions of
this paper with their comments and suggestions. AP and BEY thank Dr Abdul Fatah Amir,
Figure 11. Wallaceaphytis kikiae holotype female; habitus.
1122 A. Polaszek et al.
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Sabah Biodiversity Council, for the granting of Access Licence JKM/MBS.1000-2/2(77) under
which the field work in Sabah was carried out, and Glen Reynolds (Manager, Danum Valley
Field Studies Centre) for logistical support, and help with many other aspects of our work in
Sabah. LFs research is currently supported by a grant from the Romanian National Authority
for Scientific Research, CNCS UEFISCDI, project number PN-II-RU-TE-2012-3-0057. We
thank John Heraty, Jung-Wook Kim and Jason Mottern for making their aphelinid sequence
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... was incorporated, along with putative aphelinine genera described since their seminal publication (i.e. Indaphytis Hayat, Umairia Hayat, Wallaceaphytis Polaszek & Fusu, and Zubairia Hayat) (Polaszek et al. 2013;Hayat 2014Hayat , 2015. The new genus and species are described below, and the various characters and states are discussed in the context of establishing membership in the correct family-group. ...
... DNA extraction was undertaken using the 'non-destructive' protocol described in detail elsewhere (Polaszek et al. 2013). Unfortunately, no sequenceable DNA resulted from the extractions. ...
... n., and Wallaceaphytis kikiae Ayshford & Polaszek (all putative aphytines); and Umairia laiba Hayat, U. zeera Hayat, and Zubairia mirifica Hayat (Eutrichosomellini;Hayat 2014). Morphological character coding for I. veenakumariae and W. kikiae was performed by examining the images and descriptive taxonomy from their respective publications (Polaszek et al. 2013;Hayat 2015) (Table 1). The three eutrichosomelline genera were coded by Hayat (2014). ...
Noyesaphytis Polaszek & Woolley gen. nov. (type species Noyesaphytis lasallei Polaszek & Woolley sp. n. ) is described from Berenty, Tuléar, Madagascar. The genus differs from its closest relatives primarily in the structure of the female antenna, which has a single, elongate flagellum preceded by four anelli, the largest of which could be interpreted as a single anelliform funicle. This type of antenna is unknown in other Aphytini, but approaches the condition found in many Signiphoridae. Noyesaphytis possesses a character state that was until now thought to be an autapomorphy of Azotidae (sole genus Ablerus), being the groove in front of the propodeal spiracle. A second putative autapomorphy shared by Azotidae and Signiphoridae, and also Noyesaphytis, is the presence of anterior projections on the metasomal sterna. However, in Azotidae and Signiphoridae these are narrow, whereas as they are broader in Noyesaphytis. The form of the wing is consistent with Aphytini, although lacking a linea calva. The presumed male of Noyesaphytis lasallei has an antennal structure completely unknown in Aphelinidae, with a 1-segmented clava preceded by an extremely elongate single funicle, and four anelli. Differences between the female and male are discussed, some of which could indicate that the male might eventually be shown to belong to a different species, although the species are undoubtedly congeneric, despite the striking difference in antennal structure which is common in Aphelinidae. The male genitalia also suggest Aphytini. Based on a phylogenetic analysis of 50 morphological characters, we provisionally place Noyesaphytis in Aphytini pending the results of a forthcoming phylogenomic analysis. The new genus is named for its collector, John Noyes (NHM, London), and the new species is named after the late John La Salle.
... Genomic DNA extraction was undertaken from 10 specimens using the protocol in Polaszek et al. [10] and Cruaud et al. [11], which leaves the sclerotized parts of the specimen intact. Specimens were then critical point dried and card-mounted, with selected individuals then dissected and mounted in Canada balsam on microscope slides. ...
Full-text available
A new species of encyrtid wasp, Ooencyrtus pitosina Polaszek, Noyes & Fusu sp. n., (Hymenoptera: Encyrtidae: Encyrtinae) is described as a gregarious parasitoid in the eggs of the endemic Samoan swallowtail butterfly Papilio godeffroyi (Lepidoptera: Papilionidae) in the Samoan archipelago. It is described here because it is an important natural enemy of this butterfly, and to facilitate identification for future work with this parasitoid and its host.
... Genomic DNA was extracted from single, whole specimens using a non-destructive genomic DNA extraction protocol developed by Chao-Dong Zhu, John Noyes, and others at the Natural History Museum, London [17]. Occasionally 2-3 specimens were extracted together when known to be conspecific. ...
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The genus Dirphys Howard 1914 syn. n. is synonymized with Encarsia Förster, and treated as a species-group of Encarsia, referred to henceforth as the Encarsia mexicana species-group. The monophyly of Encarsia is discussed in relation to Dirphys. The new synonymy is based on phylogenetic analyses of the nuclear ribosomal 28S-D2 gene region (43 taxa, 510 bp). The Encarsia mexicana species-group is recovered as strongly monophyletic within Encarsia. All species of the Encarsia mexicana species-group are revised. The group includes six previously described species, and fourteen newly described species. All species are described (or redescribed) and illustrated. Detailed distributional data, and, where available, plant associate and host records are provided for all species. Encarsia myartsevae Kresslein and Polaszek nom. nov. is here proposed as a replacement name for Encarsia mexicana Myartseva, now preoccupied by Encarsia mexicana (Howard). A dichotomous identification key, supplemented by an online multiple-entry key, is provided for all species.
... Parasitoids emerging in the glass tubes of the parasitoid emergence boxes were collected and placed in 80% ethanol and stored at −20 °C for morphological identification studies. Slide mounting of parasitoids was carried out according to the method specified in Polaszek et al. (2014). Identification of the parasitoids was conducted using the morphological characters and identification keys specified in Huang and Polaszek (1998). ...
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In this study, the distribution of the Ficus whitefly, Singhiella simplex (Singh) (Hemiptera: Aleyrodidae), and its natural enemies in the Western Mediterranean Region of Turkey were investigated. For this purpose, the sampling was made from trees of Ficus spp. during Aug, Sep, and Oct when the pest population was at its peak in the various districts within the Antalya province in 2018 and 2019. In addition, the rate of natural parasitism in the sampling periods also was determined. To determine the dispersal and parasitoids of S. simplex, at least 100 branches were collected from Ficus trees in each district, the Ficus trees were checked visually for the determination of the predators. The results showed that Ficus whitefly is dispersed in all the districts within the Antalya province. Encarsia protransvena Viggiani (Hymenoptera: Aphelinidae) has been identified as the parasitoid of the Ficus whitefly in Antalya and its districts, wereas the highest natural parasitism rate was found to be 32.88% and 21.66% in Oct 2018 and 2019, respectively, across the sampling mo. Chrysoperla mutata (McLachlan) (Neuroptera: Chrysopidae), Semidalis aleyrodiformis (Stephens) (Neuroptera: Coniopterygidae), Conwentzia psociformis (Curtis) (Neuroptera: Coniopterygidae), Conwentzia sp. (Neuroptera: Coniopterygidae), Oenopia conglobata (L.) (Coleoptera: Coccinellidae), and Serangium parcesetosum Sicard (Coleoptera: Coccinellidae) species were determined as the predators. The results obtained in the study may contribute to the control of S. simplex by using its natural enemies. En este estudio, se investigó la distribución de la mosca blanca del ficus, Singhiella simplex (Singh) (Hemiptera: Aleyrodidae), y sus enemigos naturales en la región mediterránea occidental de Turquía. Para tal efecto, el muestreo se realizó a partir de árboles de Ficus spp. durante agosto, septiembre y octubre, cuando la población de plagas alcanzó su punto máximo en los diversos distritos de la provincia de Antalya en el 2018 y 2019. Además, también se determinó la tasa de parasitismo natural en los períodos de muestreo. Para determinar la dispersión y parasitoides de S. simplex se recolectaron al menos 100 ramas de árboles de Ficus en cada distrito, los árboles de Ficus fueron revisados visualmente para la determinación de los depredadores. Los resultados mostraron que la mosca blanca del ficus está dispersa en todos los distritos dentro de la provincia de Antalya. Encarsia protransvena Viggiani (Hymenoptera: Aphelinidae) ha sido identificada como el parasitoide de la mosca blanca del ficus en Antalya y sus distritos, donde se encontró que la tasa de parasitismo natural más alta del 32,88% y el 21,66% en octubre de 2018 y 2019, respectivamente. Se determinó Chrysoperla mutata (McLachlan) (Neuroptera: Chrysopidae), Semidalis aleyrodiformis (Stephens) (Neuroptera: Coniopterygidae), Conwentzia psociformis (Curtis) (Neuroptera: Coniopterygidae), Conwentzia sp., Oenopia conglobata (L.) (Coleoptera: Coccinellidae), y Serangium parcesetosum Sicard (Coleoptera: Coccinellidae) como depredadores. Los resultados obtenidos en el estudio pueden contribuir al control de S. simplex mediante el uso de sus enemigos naturales.
... Samples were removed from the tubes and gently blotted dry on clean lint-free tissue paper. The samples were then subjected to DNA extraction using the DNeasy Blood and Tissue kit (Qiagen, UK) following the manufacturer's protocol, amended to incorporate an overnight incubation at 55 °C once the buffer ATL and proteinase K had been added to the tissue (Polaszek et al. 2014). Subsequent steps were as prescribed by the manufacturer. ...
Pomacea canaliculata, commonly known as Golden Apple Snail (GAS), an invasive snail, has successfully invaded ecosystems outside its native ranges with negative impacts being reported. GAS was reported in Kenya in 2020 invading one of the largest rice-producing schemes, the Mwea irrigation scheme. Delimiting surveys were conducted in five major rice production schemes in Kenya to establish the boundary of spread since its first report and to help in the management and development of quarantine strategies to limit the spread of this pest within the scheme and other risk areas. In addition, an Ensemble model approach was used to model the potential distribution of GAS in Eastern Africa. Over 80% of the Mwea scheme’s sections were infested with GAS along the river and irrigation channel gradient from the initial infestation point (Ndekia). The mean adult/m2 and egg clutches count was 8.4 and 7.7, respectively. Significant difference was observed in number of adults/m2 and egg masses among the sections; X2(7) = 138.69, p< 0.001and X 2(7) = 114.17, p < 0.001, respectively. The survey did not find any adults or eggs of GAS in the Ahero, Bura, Hola and West Kano rice schemes. Most of the areas in Kenya were suitable for GAS invasion, though with significant variations across the country. At a regional level, the countries with the highest suitability for GAS were Malawi, Madagascar and Uganda. Mozambique, Tanzania and Ethiopia also had areas of high suitability, but these were more concentrated in specific areas in each of the countries. In comparison, suitability across Sudan and Somalia was very low. Strict quarantine measures should be instituted and implemented to curb not just the spread of GAS in Kenya but entry into uninvaded regions.
... Only puparia from which no adult had emerged were selected for DNA extraction and to ensure that there would be no anomalous image data due to the presence of moulting sutures. Genomic DNA was extracted from each specimen following the protocol described by Polaszek et al. (2013). Immediately after removal from the enzyme/buffer mix, all puparia were processed for slide mounting. ...
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The Bemisia tabaci species-complex is a group of tropical-subtropical hemipterans, some species of which have achieved global distribution over the past 150 years. Several species are regarded currently as among the world’s most pernicious agricultural pests, causing a variety of damage types via direct feeding and plant-disease transmission. Long considered a single variable species, genetic, molecular and reproductive compatibility analyses have revealed that this “species” is actually a complex of between 24 and 48 morphologically cryptic species. However, determinations of which populations represent distinct species have been hampered by a failure to integrate genetic/molecular and morphological species-diagnoses. This, in turn, has limited the success of outbreak-control and eradication programmes. Previous morphological investigations, based on traditional and geometric morphometric procedures, have had limited success in identifying genetic/molecular species from patterns of morphological variation in puparia. As an alternative, our investigation focused on exploring the use of a deep-learning convolution neural network (CNN) trained on puparial images and based on an embedded, group-contrast training protocol as a means of searching for consistent differences in puparial morphology. Fifteen molecular species were selected for analysis, all of which had been identified via DNA barcoding and confirmed using more extensive molecular characterizations and crossing experiments. Results demonstrate that all 15 species can be discriminated successfully based on differences in puparium morphology alone. This level of discrimination was achieved for laboratory populations reared on both hairy-leaved and glabrous-leaved host plants. Moreover, cross-tabulation tests confirmed the generality and stability of the CNN discriminant system trained on both ecophenotypic variants. The ability to identify B. tabaci species quickly and accurately from puparial images has the potential to address many long-standing problems in B. tabaci taxonomy and systematics as well as playing a vital role in ongoing pest-management efforts.
... Eight Telenomus nizwaensis individuals from Oman (4 females, 4 males) were subjected to "non-destructive" DNA extraction. Genomic DNA was extracted using established protocols [12,13], which leave the sclerotized parts of the specimen intact. Specimens were then critical point dried and card-mounted, with selected individuals then dissected and mounted in Canada balsam on microscope slides, and others gold-palladium coated for SEM examination. ...
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The pomegranate butterfly Deudorix (= Virachola) livia is the major pest of pomegranate, a crop of economic importance, in Oman. A species of parasitoid wasp in the hymenopteran family Scelionidae is responsible for high levels of mortality of its eggs. This wasp is described herein as Telenomus nizwaensis Polaszek sp. n., based on morphology and DNA sequence data. T. nizwaensis is currently known only from D. livia, which is also a pest of economic importance on other crops in North Africa, the Arabian Peninsula, and the Mediterranean. We summarise current knowledge of T. nizwaensis life-history and its potential to provide biological pest control.
... Samples were removed from the tubes and gently blotted dry on clean lint-free tissue paper. The samples were then subjected to DNA extraction using the DNeasy Blood and Tissue kit (Qiagen, UK) following the manufacturer's protocol, amended to incorporate an overnight incubation at 55 °C once the buffer ATL and proteinase K had been added to the tissue (Polaszek et al. 2014). Subsequent steps were as prescribed by the manufacturer. ...
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Following reports of an invasive snail causing crop damage in the expansive Mwea irrigation scheme in Kenya, samples of snails and associated egg masses were collected and sent to CABI laboratories in the UK for molecular identification. DNA barcoding analyses using the cytochrome oxidase subunit I gene gave preliminary identification of the snails as Pomacea canaliculata, widely considered to have the potential to be one of the most invasive invertebrates of waterways and irrigation systems worldwide and which is already causing issues throughout much of south-east Asia. To the best of our knowledge, this is the first documented record of P. canaliculata in Kenya, and the first confirmed record of an established population in continental Africa. This timely identification shows the benefit of molecular identification and the need for robust species identifications: even a curated sequence database such as that provided by the Barcoding of Life Data system may require additional checks on the veracity of the underlying identifications. We found that the egg mass tested gave an identical barcode sequence to the adult snails, allowing identifications to be made more rapidly. Part of the nuclear elongation factor 1 alpha gene was sequenced to confirm that the snail was P. canaliculata and not a P. canaliculata/P. maculata interspecies hybrid. Given the impact of this species in Asia, there is need for an assessment of the risk to Africa, and the implementation of an appropriate response in Kenya and elsewhere to manage this new threat to agriculture and the environment.
... Genomic DNA was extracted following the protocol in Polaszek et al. (2013), which leaves the sclerotised parts of the specimen intact. The specimen was subsequently slidemounted in Canada balsam. ...
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The Chinese species of the varius group of Coccophagus Westwood (Hymenoptera: Aphelinidae) are reviewed, including described species Coccophagus albifuniculatus (Huang), C. anchoroides (Huang), C. caudatus (Huang), C. dilatatus (Huang), C. equifuniculatus (Huang), C. fumadus Hayat (new record for China), C. lii (Huang), C. pellucidus (Huang), and one new species, Coccophagus yunnana Wang, Huang & Polaszek sp. nov. A revised key to the females of the Chinese species of the varius group of Coccophagus is provided. Mitochondrial (CO1) and nuclear ribosomal (28S-D2) partial sequences were obtained successfully for C. fumadus.
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A new, biparental species of the genus Encarsia Förster (Hymenoptera: Aphelinidae), E. hera Lahey & Andreason, sp. nov., is characterized based on morphological and molecular data. The parasitoid was reared from the puparia of its host, an undescribed species of the grass-feeding aleyrodine genus Aleurocybotus Quaintance & Baker (Hemiptera: Aleyrodidae) collected in Gainesville, Florida. The same whitefly is newly recorded from Charleston, South Carolina, where it is a pest of ornamental Muhly grass [Muhlenbergia capillaris (Lam.) Trin. (Poaceae)]. A phylogenetic analysis based on a fragment of 28S ribosomal DNA in 34 Encarsia species placed E. hera, sp. nov., within the E. luteola-group, a result concordant with its morphology. A key to the Encarsia species reared from Aleurocybotus is provided.
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Although much biological research depends upon species diagnoses, taxonomic expertise is collapsing. We are convinced that the sole prospect for a sustainable identification capability lies in the construction of systems that employ DNA sequences as taxon 'barcodes'. We establish that the mitochondrial gene cytochrome c oxidase I (COI) can serve as the core of a global bioidentification system for animals. First, we demonstrate that COI profiles, derived from the low-density sampling of higher taxonomic categories, ordinarily assign newly analysed taxa to the appropriate phylum or order. Second, we demonstrate that species-level assignments can be obtained by creating comprehensive COI profiles. A model COI profile, based upon the analysis of a single individual from each of 200 closely allied species of lepidopterans, was 100% successful in correctly identifying subsequent specimens. When fully developed, a COI identification system will provide a reliable, cost-effective and accessible solution to the current problem of species identification. Its assembly will also generate important new insights into the diversification of life and the rules of molecular evolution.
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Chalcidoidea (Hymenoptera) are extremely diverse with more than 23,000 species described and over 500,000 species estimated to exist. This is the first comprehensive phylogenetic analysis of the superfamily based on a molecular analysis of 18S and 28S ribosomal gene regions for 19 families, 72 subfamilies, 343 genera and 649 species. The 56 outgroups are comprised of Ceraphronoidea and most proctotrupomorph families, including Mymarommatidae. Data alignment and the impact of ambiguous regions are explored using a secondary structure analysis and automated (MAFFT) alignments of the core and pairing regions and regions of ambiguous alignment. Both likelihood and parsimony approaches are used to analyze the data. Overall there is no impact of alignment method, and few but substantial differences between likelihood and parsimony approaches. Monophyly of Chalcidoidea and a sister group relationship between Mymaridae and the remaining Chalcidoidea is strongly supported in all analyses. Either Mymarommatoidea or Diaprioidea are the sister group of Chalcidoidea depending on the analysis. Likelihood analyses place Rotoitidae as the sister group of the remaining Chalcidoidea after Mymaridae, whereas parsimony nests them within Chalcidoidea. Some traditional family groups are supported as monophyletic (Agaonidae, Eucharitidae, Encyrtidae, Eulophidae, Leucospidae, Mymaridae, Ormyridae, Signiphoridae, Tanaostigmatidae and Trichogrammatidae). Several other families are paraphyletic (Perilampidae) or polyphyletic (Aphelinidae, Chalcididae, Eupelmidae, Eurytomidae, Pteromalidae, Tetracampidae and Torymidae). Evolutionary scenarios discussed for Chalcidoidea include the evolution of phytophagy, egg parasitism, sternorrhynchan parasitism, hypermetamorphic development and heteronomy.
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Comparative analysis of molecular sequence data is essential for reconstructing the evolutionary histories of species and inferring the nature and extent of selective forces shaping the evolution of genes and species. Here, we announce the release of Molecular Evolutionary Genetics Analysis version 5 (MEGA5), which is a user-friendly software for mining online databases, building sequence alignments and phylogenetic trees, and using methods of evolutionary bioinformatics in basic biology, biomedicine, and evolution. The newest addition in MEGA5 is a collection of maximum likelihood (ML) analyses for inferring evolutionary trees, selecting best-fit substitution models (nucleotide or amino acid), inferring ancestral states and sequences (along with probabilities), and estimating evolutionary rates site-by-site. In computer simulation analyses, ML tree inference algorithms in MEGA5 compared favorably with other software packages in terms of computational efficiency and the accuracy of the estimates of phylogenetic trees, substitution parameters, and rate variation among sites. The MEGA user interface has now been enhanced to be activity driven to make it easier for the use of both beginners and experienced scientists. This version of MEGA is intended for the Windows platform, and it has been configured for effective use on Mac OS X and Linux desktops. It is available free of charge from
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Aphelinidae are all insect parasitoids, and most species are associated with nymphal stages of Homoptera: Sternorrhyncha, either as primary parasitoids or hyperparasitoids. The occurrence of egg parasitism in aphelinids has been recorded in eight of the 38 valid genera and these records are reviewed; it is particularly common in the genus Centrodora, which is shown to be the most polyphagous in the family. One species, C. darwini (Girault), is given special attention because of its occurrence in three recent surveys for biological control agents of crop pests. It is briefly redescribed, diagnosed, and shown to be the most polyphagous aphelinid known. A checklist of Australian Centrodora spp. is given, including the new combination Centrodora grotiusi (Girault) comb. n. The purported evidence for the classification of certain Encarsia spp. associated with eggs of Lepidoptera as ‘heterotrophic parasitoids’ is re-examined and dismissed.
The genera of Aphelininae (Aphelinidae) are reviewed on a worldwide basis. Identification keys and a phylogenetic hypothesis are presented for 16 genera, of which four are new (Mashimaro n.g., Neophytis n.g., Punkaphytis n.g., Saengella n.g.) and Paraphytis is resurrected. Newly described species are Mashimaro hawksi n.sp., Mashimaro lasallei n.sp., Neophytis myartsevae n.sp., Neophytis munroi n.sp., Punkaphytis erwini n.sp. and Punkaphytis hayati n.sp. New combinations from Aphytis include Neophytis melanosticus (Compere) and N. dealbatus (Compere), and Paraphytis acutaspidis (Rosen & DeBach), P. angusta (Compere), P. anomala (Compere), P. argenticorpa (Rosen & DeBach), P. australiensis (DeBach & Rosen), P. benassyi (Fabres), P. breviclavata (Huang), P. capillata (Howard), P. ciliata (Dodd), P. cochereaui (DeBach & Rosen), P. costalimai (Gomes), P. densiciliata (Huang), P. fabresi (DeBach & Rosen), P. haywardi (De Santis), P. hyalinipennis (Rosen & DeBach), P. maculatipennis (Dozier), P. maculata (Shafee), P. mandalayensis Rosen & DeBach, P. nigripes (Compere), P. noumeaensis (Howard), P. obscura (DeBach & Rosen), P. peculiaris (Girault), P. perplexa (Rosen & DeBach), P. transversa (Huang), P. vittata (Compere) (revived status) and P. wallumbillae (Girault). A parsimony analysis of 50 morphological characters for 54 species in 21 genera, including four outgroups (Coccophagus, Eunotiscus, Euryishomyia and Eriaphytis), resulted in three equally parsimonious trees supporting monophyly of all genera except Neophytis. Neophytis was paraphyletic to a monophyletic Aphytini but is monophyletic in unpublished molecular analyses. Three tribes are recognised (Aphelinini, Aphytini and Eutrichosomellini). The questionable inclusion of Eretmocerus (Eretmocerini) within Aphelininae is discussed.
The most profitable ways of collecting chalcids are discussed, namely by sweeping, suction sampler, beating, pyrethrum spray, rearing, Malaise traps, yellow pan traps, suction traps, pitfall traps and extraction from leaf litter or grass tussocks. Methods of preserving chalcids are also outlined, with particular emphasis on storing all unmounted material dry rather than in alcohol. The techniques of sweeping, card mounting specimens and slide preparation are described in detail.
The skeletomusculature of the mesofurcal–mesopostnotal complex is surveyed within the Chalcidoidea. Four internal character systems are assessed for their phylogenetic significance: the mesofurcal bridge, the structure and position of the furcal–laterophragmal muscle, the structure of the lateral arms of the mesofurca, and the supporting structures for the interfurcal muscles. Among Hymenoptera, Chalcidoidea are unique in having the furcal–laterophragmal muscle attached along the entire length of the laterophragmal apodeme. Also the furcal–laterophragmal muscle originates medial to the junction of the mesofurcal bridge and lateral mesofurcal arm in most Chalcidoidea. Mymarommatidae do not share either of these apomorphic states with Chalcidoidea. Within Chalcidoidea, apomorphic character states were found in each of Aphelinidae, Encyrtidae, Eulophidae, Mymaridae, Rotoitidae, Signiphoridae, Tanaostigmatidae and Trichogrammatidae. For taxa classified as Aphelinidae, the plesiomorphic complement of structures and muscle attachments is retained in Eriaphytinae and Eriaporinae. The mesofurcal bridge is considered to have been lost at least twice in each of Aphelininae and Coccophaginae. Similar interfurcal processes, resulting from loss of the mesofurcal bridge, support the monophyly of Aphelininae (Aphelinini, Aphytini and Eutrichosomellini). Azotinae are placed as the sister group of Aphelininae because of a similar lateral origin of the laterophragmal muscle and the shape of the mesofurcal arms. Other than loss of the mesofurcal bridge, no character states were shared by Azotinae and Coccophaginae. Coccophaginae (Coccophagini and Pteroptricini) are regarded as monophyletic based on the loss of the mesofurcal bridge, the peculiar shape of the mesofurca, and a unique modification of the laterophragmal muscle. Euxanthellus is removed from synonomy with Coccophagus and may be best treated as a separate tribe of Coccophaginae based on the shape of the lateral mesofurcal arms and the presence of a mesofurcal bridge. The shape of the mesofurca suggests a monophyletic grouping of Cales, Eretmocerus and Trichogrammatidae that could render Aphelinidae paraphyletic.
A data set consisting of twenty-eight anatomical characters scored for twenty-eight terminal taxa representing the world fauna of Signiphoridae was analysed using parsimony and compatibility methods. The Coccophaginae (Aphelinidae) and the Azotinae (Aphelinidae) were used as outgroups to establish polarity of character state changes. Relationships of Signiphoridae to other Chalcidoidea are discussed. Several multistate characters were treated in the parsimony analyses either as unordered or as ordered into transformation series using additive binary coding, which in some cases drastically reduced the number of equally parsimonious solutions. Monophyly of Signiphoridae is supported by seven synapomorphies. Four genera, Chartocerus, Thysanus, Clytina and Signiphora, are recognized within Signiphoridae based on synapomorphies. Rozanoviellasyn.n. and Kerrichiellasyn.n. are synonymized under Signiphora. Species of Signiphora are further assigned to four species groups, three of which are demonstrably monophyletic. Nine species or subspecies are transferred to Chartocerus from Signiphora (australicuscomb.n., australiensiscomb.n., australiensis orbiculatus comb.n., beethovenicomb.n., corvinuscomb.n., funeraliscomb.n., reticulatacomb.n., ruskinicomb.n., thusanoidescomb.n.), one species to Thysanus from Signiphora (melancholicuscomb.n.), and one species to Signiphora from Kerrichiella (coleoptratuscomb.n.). A key to genera of Signiphoridae and species groups of Signiphora is presented. A diagnosis, relevant nomenclatural history, and a list of included species are given for each genus and species group, and the biology and distribution of each is summarized.