Cytogenetics of New Zealand blackflies of the genus Austrosimulium (Diptera: Simuliidae) 1. The cytogenetics of Austrosimulium australense

Abstract

This study undertakes a cytogenetic analysis of the New Zealand blackfly species Austrosimulium australense (Schiner). The principles of such an approach are outlined, and previous taxonomic studies of the genus Austrosimulium, in particular the taxonomic position of A. australense, are discussed. Populations from North Island localities covering a wide area were sampled and analysed for polymorphisms in the polytene chromosomes, taken from salivary glands of larvae. In all, 1018 larvae from 49 sites were analysed. A polytene chromosome map of A. australense is presented, with details of chromosomal inversions found in salivary gland cells. Three geographical zones are designated, according to the incidence of certain chromosomal polymorphisms.     

Wing and haltere venation in larvae of the Austrosimulium (Austrosimulium) australense group from New Zealand (Diptera: Simuliidae)

The venation pattern found in the developing wing and haltere buds of larvae is compared with the adult venation in the Austrosimulium (Austro-simulium) australense group of Simuliidae.     

Life history stages and taxonomy of Austrosimulium (Austrosimulium) tillyardianum (Diptera: Simuliidae)

The male, female, pupa, 9th‐instar larva, and egg of Austrosimulium (Austrosimulium) tillyardianum Dumbleton, 1973, are redescribed, and larval instars one to eight are described and morphologically differentiated for the first time. The relationship of A. (A.) tillyardianum to other New Zealand species is briefly reviewed.     

Colonization of perspex strips by larvae of Austrosimulium bancrofti (Taylor) near Ipswich,Queensland

Larvae of the Taxon Austrosimulium bancrofti (Taylor) were collected from perspex strips placed in rapids of the Brisbane River during January (summer) and October (spring). Separation of larvae according to the mixture approach to clustering demonstrated that: a) October cluster I larvae were larger and underwent an additional cluster to those collected in January, b) there was an additional cluster of larvae in each month, which correlated with the percentage of A. bancrofti Ipswich B that were identified cytologically, and c) greater numbers of later cluster larvae were collected from the strips than early clusters.Multivariate analysis demonstrated that distance from the lip of the rapids, strip texture and water velocity had a significant effect on the number of larvae colonizing perspex strips. Numbers increased with: a) decreasing distance from the lip of the rapids, b) increasing surface area of roughened strips and c) water velocity in the range 1.00 < X 1.38ms^–1. The influence of adult female oviposition behaviour, larval behaviour, sampling time and technique are discussed in relation to the parameters that influenced the colonization of artificial substrates.     

The water velocity preferences of Austrosimulium furiosum and Simulium ornatipes (Diptera: Simuliidae) larvae,and the implications for micro-habitat partitioning

The aquatic larvae of two simuliid species, Austrosimulium furiosum (Skuse) and Simulium ornatipes Skuse, which often occur together in Victorian streams, were shown in laboratory experiments to have preferences for different water velocities: larvae of A. furiosum preferred water velocities of 0.2–0.3 m s^–1, and S. ornatipes preferred water velocities of 0.9–1.3 m s ^–1 . Final instar larvae of both species selected slow water speeds of less than 0.25 m s ^–1 prior to pupation. Flow patterns around a cylinder in a laboratory stream were mapped, and the distribution of A. furiosum larvae within the wake, paired vortices and horseshoe vortex was recorded. Larvae selected areas with suitable water velocities and aligned with the flow, providing flow visualization of micro-current speed and direction. The potential for micro-habitat partitioning is discussed in relation to benthic sampling strategies.     

Different colonization patterns of two closely related stream insects (Austrosimulium spp.) following disturbance

SUMMARY.

• 1 In many streams, blackfly larvae rapidly colonize scoured surfaces, then decline in numbers as other species are still increasing. Such a colonization pattern may be generated by species that seek disturbed substrata because they prefer scoured surfaces having low epilithon cover (i.e. they are opportunists) or because of lowered faunal densities (i.e. they are fugitives), but this has rarely been tested.
• 2 In the Acheron River, south-eastern Australia, the larvae of the blackfly Austrosimulium torrentium show the typical pattern of increase and decline whereas A. victoriae shows a different pattern of more gradual increase.
• 3 An experiment showed that A. torrentium, consistent with its colonization pattern, responded strongly to treatments that mimicked the effects of disturbance; it colonized bricks with few other animals present regardless of epilithon cover and therefore is potentially a fugitive species. A. victoriae did not respond convincingly to treatments, which is also consistent with its colonization pattern.
• 4 Blackfly larvae of both species preferentially colonized bricks that were in fast-moving water. Hence, A. torrentium only responded to the densities of other fauna when bricks occurred in currents greater than approximately 30 cm s^?1.
• 5 Whilst epilithon cover did not affect abundance of blackfly larvae, it did affect attachment positions. In one experimental treatment, epilithon was reduced on one half of the top surfaces of bricks. More than 50% of larvae on these bricks were attached to the epilithon-reduced halves.
• 6 Our results show that functional feeding or taxonomic groups do not necessarily contain ecologically equivalent taxa; colonization patterns may provide a more useful way of categorizing species for the purposes of modelling these assemblages.

     

Cytogenetics of New Zealand blackflies of the genus Austrosimulium (Diptera: Simuliidae) II. Heterozygote deficiency and non-random association of inversion heterozygotes

We present population cytogenetic data from an analysis of polytene chromosomes of individuals of the common New Zealand blackfly Austrosimulium australense (Schiner). These data show that, in an area of the central North Island of New Zealand (Tarawera), there are, firstly, a number of abrupt changes in inversion polymorphisms between this area and adjacent collection sites. Secondly, there is non-random association of heterozygous inversion pairs in samples from the Tarawera area. Thirdly, we present evidence of a deficiency of heterozygotes for a number of inversions found in this area. We conclude that this may provide evidence for the existence of cryptic species within our samples. We discuss this possibility in terms of the morphological and genetical changes which are likely to occur at speciation, and briefly discuss the mounting evidence for the ubiquity of cryptic species in Diptera.Publication No. 10 from the Evolutionary Genetics Laboratory, University of Auckland.     

An adaptive bivariate kernel smoothing method for determining instars of Austrosimulium tillyardianum (Diptera: Simuliidae) larvae

1. In insects, instar determination is generally based on the frequency distribution of sclerotised body part measurements. Commonly used univariate methods, such as histograms and univariate kernel smoothing, are not sufficient to reflect the distribution of the measurements, because development of sclerotised body parts is multidimensional. 2. This study used an adaptive bivariate kernel smoothing method, based on 10 pairs of separating variables, to differentiate instars of Austrosimulium tillyardianum (Diptera: Simuliidae) larvae in two‐dimensional space. A variable bandwidth matrix was used and separation lines between instars were defined. Using the Crosby growth ratio, Brooks' rule and the new standard recently proposed, larvae were separated into nine instars. It was found that, using the bivariate kernel smoothing method, the clustering accuracy and determination of separation lines as instar class limits were higher than those associated with the univariate kernel smoothing method. With the exceptions of the paired separating variables, head capsule length and antennal segment 3 length (AS3L), the mean probabilities of correct classifications was > 85%. The pair of separating variables that yielded the greatest classification accuracy comprised mandible length (ML) and AS3L, which had mean probabilities of 0.8984. The clustering accuracy was higher for early‐ and late‐instar larvae, but lower for instars 6 and 7. The adaptive bivariate kernel smoothing method was better than univariate methods for instar determination, especially in the detection of divisions between instars and identification of a larval instar.     

Further observations on the life cycle and vectors of the haemosporidian Leucocytozoon tawaki and its transmission to the Fiordland crested penguin

Abstract

Hepatic and renal schizonts are described from Fiordland crested penguin (Eudyptes pachyrhynchus) chicks, and renal and splenic schizonts from mature birds. Ookinetes, oocysts, and sporozoites are described from the primary vector, Austrosimulium ungulatum. Penguin chicks are infected when 4–6 weeks old. Some observations are recorded on the biology of the vector in association with the birds.     

Notes on New Zealand mammals 9: Long‐tailed bat (Chalinolobus tuberculatus) chased by morepork (Ninox novaeseelandiae)

Blood smears from 43 species of bird, mainly from the South Island of New Zealand, were examined for haematozoon parasites. An undescribed species of Leucocytozoon was discovered in the Fiordland crested penguin, Eudyptes pachyrhynchus. It is assigned the name Leucocytozoon tawaki, and the gametocytes are described. Preliminary observations of its development in Austrosimulium austra‐lense, A. dumbletoni, and A. ungulatum are discussed, and the ookinete, oocyst, and sporozoite stages within these hosts are briefly described.     

Polytene chromosomes in pupal and adult blackflies (Diptera: Simuliidae)

A number of pupal and adult tissues of eight Australian blackfly species representing three genera, Austrosimulium, Cnephia and Simulium, were examined for the presence of polytene chromosomes. Banded polytene chromosomes were found in malpighian tubules, hind gut, fat body, and ovary, but only those from the malpighian tubules of female adults and pupae were of good quality. A detailed comparison of polytene chromosomes from larval salivary glands and adult malpighian tubules was made in S. ornatipes and, to a limited extent, in S. melatum. The banding patterns of chromosomes from both tissues were found to be identical with minor differences in puffing patterns in S. ornatipes and chromocenter characteristics in S. melatum. A survey of the remaining six species shows five of them to have malpighian chromosomes suitable for detailed cytological analysis. Simultaneous studies of larval, pupal and adult polytene chromosome systems offer a novel approach to the analysis of population problems in blackflies. The ability to recognise sibling species in adults also has potential practical significance in efforts to control vectors of onchocerciasis.     

Factors influencing silhouette trap captures of the blackfly Austrosimulium bancrofti in Queensland, Australia

Three silhouette traps of novel design were used to investigate the trap finding ability of female Austrosimulium bancrofti (Taylor) (Diptera: Simuliidae) in southeast Queensland. The traps were 'upright', 'elongate' or 'square' in shape. Five highly significant factors: hour after sunrise, temperature, relative humidity, trap shape and distance between traps, were included in a model which generated fitted estimates of flies captured under defined conditions. The rate of increase and the level of maximum capture rate indicated that the elongate trap captured the highest numbers of blackflies. An interaction between hour after sunrise and temperature suggests a strong endogenous modification of trap finding activity. Numbers captured increased with increasing distance between traps and decreasing relative humidity. The importance of these results in understanding behaviour of A. bancrofti is discussed.     

Tyrosinase inhibitory activities of the compounds isolated from Neolitsea aciculata (Blume) Koidz.

This study was designed to investigate the mushroom tyrosinase inhibitory activity for the constituents isolated from Neolitsea aciculata. The stems of N. aciculata was extracted with aqueous ethanol and subjected to chromatographic separation, which led to the isolation of 11 compounds: methyl linoleate (1), catechin (2), epicatechin (3), afzelin-7-O-glucopyranoside (4), 2′,3′-di-(p-coumaroyl)afzelin (5), 2′-p-coumaroylafzelin (6), feruloyl tyramine (7), β-sitosterol (8), daucosterol (9), oleic acid (10), and trilaurin (11). Their structures were elucidated on the basis of spectroscopic studies as well as by comparison with the data available in the literature. Among these isolates, compounds 5 and 6 were identified as potent mushroom tyrosinase inhibitors with IC₅₀ values of 0.067 and 0.080 mM, respectively. The inhibition kinetics, analysed by Lineweaver–Burk plots, indicated that compounds 5 and 6 are competitive tyrosinase inhibitors when using l-tyrosine as a substrate. Notably, compounds 1–11 were isolated for the first time from this plant. These results provide evidence that this plant might be a potential source of anti-melanogenesis agents.     

Folsomia (Collembola: Isotomidae) of China. II. heterocellata group

An annotated list of species of the Folsomia heterocellata group recorded in China is given. Folsomia cephalota n. sp., F. dorsomediaseta n. sp., F. longiseta n. sp., F. manubriopilosa n. sp., F. longdeica n. sp., and F. plenosensilla n. sp. are characterized by having a simple maxillary palp, four prelabral setae and no eyes. They differentiate mainly by characteristics of the furca. We report other species of the group which are recorded for the first time in China: Folsomia tianshanica Martynova, 1969, F. pseudovtorovi Potapov & Gulgenova, 2013, and F. pseudodecemoculata Stebaeva, 1971. This excludes F. wanxianensis Ji, Ding & Chen, 2007, for which we provide morphological remarks based on the type material. The group is mainly distributed in the central part of China (Chongqing, Gansu, southern part of Inner Mongolia, Ningxia and Shaanxi provinces). A key to all blind species with a simple maxillary palp is given.     

New antibacterial pentacyclic triterpenes from Myricaria elegans Royle. (tamariscineae)

Two new pentacyclic triterpenes eleganene-A (1) and eleganene-B (2), along with four known pentacyclic triterpenes betulin (3), ursolic acid (4), erythrodiol (5) and corosolic acid (6) were isolated from the aerial parts of Myricaria elegans. These compounds exhibited significant antibacterial activity. The structure of compounds 1 and 2 were deduced on the basis of their spectral analysis.     

A comparison of Dichodontium flavescens (Dicks.) Lindb. and D. pellucidum (Hedw.) Lindb. (Bryopsida)

Abstract

Fifty-three herbarium samples of the genus Dichodontium, originating from 15 countries in Europe and North America, were examined to critically assess new distinguishing characters between D. flavescens and D. pellucidum. The examination, including SEM, revealed some new characteristics in the gametophyte, notably leaf shape, leaf length: width ratio, and width of the nerve, permitting accurate determinations even on sterile material. D. flavescens is recognized here as a distinct species.     

potamolejeunea Holtii (Spruce) nov.comb.

Abstract

Bryophytes usually have anti-feeding properties to defend against microbial and herbivore attack; however, the consumption of Haplocladium microphyllum (Hedw.) Broth. capsules by Agrotis sp. larvae is rather common in Shanghai in the spring. To test whether H. microphyllum is the only moss eaten, and why the gametophytes of H. microphyllum are not eaten, a series of quantitative experiments were carried out to understand the feeding habits of Agrotis larvae on the given moss materials at three growth stages of larval life. The results show that the larvae can feed on the capsules of six moss species to different degrees: Funaria hygrometrica Hedw., H. microphyllum, Physcomitrium sphaericum (C.F.Ludw. ex Schkuhr) Brid., Trematodon longicollis Michx., Ditrichum pallidum (Hedw.) Hampe, and Pogonatum inflexum (Lindb.) Sande Lac. The capsules of the first four species were grazed heavily by the larvae, compared with limited consumption of the latter two (D. pallidum and P. inflexum), which even induced a high mortality rate among the larvae. With the growth of the larval instar, the daily demand for moss capsules by the larvae increases gradually. The lipid content of the capsules may play an important role when the larva selects its feeding target.     

A new species of Lathyrus L. (Fabaceae) from Central Apennine (Italy)

Abstract

Lathyrus apenninus, from beech woods margins and scrub in the Central Apennine (Italy), is described as a species new to science. The relationship of L. apenninus with L. linifolius, which it had been confused with, is briefly discussed.     

Effective Cytochrome P450 (CYP) Inhibitors Isolated from Tarragon (Artemisia dracunculus)

Two effective cytochrome P450 (CYP) inhibitors were isolated from tarragon, Artemisia dracunculus. Their structures were spectroscopically identified as 2E,4E-undeca-2,4-diene-8,10-diynoic acid isobutylamide (1) and 2E,4E-undeca-2,4-diene-8,10-diynoic acid piperidide (2). Both compounds had dose-dependent inhibitory effects on CYP3A4 activity with IC₅₀ values of 10.0 ± 1.3 µM for compound 1 and 3.3 ± 0.2 µM for compound 2, and exhibited mechanism-based inhibition. This is the first reported isolation of effective CYP inhibitors from tarragon (Artemisia dracunculus) purchased from a Japanese market.