Triploidy in Hochstetter's frog,Leiopelma hochstetteri,from New Zealand

Abstract

Autotriploidy is described in a female of the endemic New Zealand frog Leiopelma hochstetteri. This frog was found to have 3n=33 chromosomes plus 2 supernumerary chromosomes. All the chromosomes in the karyotype of this species contained C-band heterochromatin at the centromeres. A prominent C-band was found to be associated with a secondary constriction on chromosome no. 7. The supernumerary chromosomes in this species appear to be mitotically stable and contain C-band heterochromatin at the centromeres. From the limited data presently available, the triploid individual may have resulted from the fertilisation of a diploid egg produced when the second meiotic division had been suppressed.     

Rapid recovery of a population of the cryptic and evolutionarily distinct Hochstetter's Frog,Leiopelma hochstetteri,in a pest‐free environment

New Zealand's native frogs (genus Leiopelma) display a very high degree of endemism and belong to a distinctive and ancient evolutionary lineage. All four extant species are considered to be threatened or endangered, but the mechanisms behind their decline are poorly understood. Many of the potential factors causing population declines are confounded for the two species living on the mainland; in particular, habitat degradation and the presence of introduced mammalian pests have been difficult to disentangle in the past. Here we present data on a population of Hochstetter's Frog (Leiopelma hochstetteri) living in a pest‐free mainland sanctuary, the 3363‐ha Maungatautari Ecological Island. Survey data from this population shows a fourfold increase in numbers between 2009 and 2012 from a very low initial density. Our results suggest that for Hochstetter's Frog at least, and possibly the other mainland species of Leiopelma, introduced mammals are a major driving factor in population declines. Given that the frog population at Maungatautari was discovered by serendipity when planning for the pest‐free sanctuary was already well underway, this also highlights the conservation value of large pest‐free areas and ecosystem‐level management.     

Integrated control of apple pests in New Zealand 6. Incidence of European red mite,Panonychus ulmi (Koch), and its predators

Mite relationships were investigated in an orchard near Nelson from 1962 to 1974 in conjunction with a long‐term experiment into integrated control of apple pests. From 42% to 57% of the total winter eggs of Panonychus ulmi hatched over a period of 48–53 days from the last week of September until mid November. The main hatch—from 10% to 90%—took 19–20 days. Five generations occurred in each season, from October to March, although some females laid winter eggs from mid January if leaf condition became poor. Insect predators found were Stethorus bifidus, Sejanus albisignata, Arthrocnodax sp., and Xylapothrips sp. The mite Typhlodromus pyri (Phytoseiidae), the most important predator, had up to 5 generations from October to March; some females ceased to feed or oviposit from mid February onwards, and took on the characteristics of the overwintering diapause phase. With the integrated programme based on ryania, a favourable balance of P. ulmi and T. pyri was established after an initial outbreak of P. ulmi in the 1st or 2nd season, and was maintained until 1970–71, when P. ulmi outbreaks occurred on several cultivars in a season marked by hot, dry conditions in November and December. Natural control was restored the next year except on cy. Delicious, where P. ulmi remained in greater numbers for several years. With a complete commercial spray programme from 1962 to 1967 P. ulmi became resistant to all acaricides used; with the regular azinphos‐methyl programme and no acaricides in 1967–68 a population of T. pyri resistant to this insecticide emerged. The approximately × 10 resistance level did not increase appreciably, so the azinphos‐methyl dosage was reduced from 0.05% to 0.025% to allow greater survival. Several hot, dry summers followed, and commercial control of P. ulmi by T. pyri did not occur until 1973–74. A programme of low‐dosage carbaryl allowed T. pyri and S. albisignata to establish in fair numbers, but control of P. ulmi was not achieved. Experiments with mineral oils and cyhexatin have shown them to have some potential for inclusion against P. ulmi in future integrated programmes.     

The evolution of hypervariable sex and supernumerary (B) chromosomes in the relict New Zealand frog,Leiopelma hochstetteri

Chromosomes exhibiting elevated levels of differentiation are termed hypervariable but no proposed mechanisms are sufficient to account for such enhanced evolutionary divergence. Both hypervariable sex and supernumerary (B) chromosomes were investigated in the endemic New Zealand frog, Leiopelma hochstetteri, which is chromosomally polymorphic both within and between populations and has sufficiently elevated variation that different populations can be identified solely by their C-banded karyotypes. This frog is further distinguished by the univalent, female-specific W-chromosome (0W/00 sex determination) uniquely possessed by North Island populations. This sex chromosome exhibited variation in morphology, size, and heterochromatin distribution, sufficient to resolve 11 different types, including isochromosomes. Five of the 12 populations examined also had supernumerary chromosomes that varied in number (up to 15 per individual) and morphology. Specific variations seen among the hypervariable chromosomes could have resulted from heterochromatinisation, chromosome fusions, loss-of-function mutations, deletions, and/or duplications. Frogs of the same species from Great Barrier Island, however, had neither supernumeraries nor the female-specific chromosome. The 0W/00 sex chromosome system must have been derived after the isolation of Great Barrier Island from North Island populations by raised sea levels between 14 000 and 8000 years ago. Furthermore, biochemical divergence between populations is minor and therefore the chromosomal variation seen is comparatively recent in origin. The one characteristic common to all known hypervariable chromosomes is curtailment or lack of recombination. Their accelerated evolution therefore is possible via the mechanism of Muller's ratchet, either alone or in concert with other factors.     

The taxonomic identity of a population of terrestrial Leiopelma (Anura: Leiopelmatidae) recently discovered in the northern king country,New Zealand

Abstract

A terrestrial endemic frog resembling Leiopelma archeyi was discovered in the Whareorino Forest, northern King Country, New Zealand, in 1991, where it is broadly sympatric with L. hochstetteri. To clarify its taxonomic status, allozyme electrophoresis of toe tissue was used to compare it genetically with four other populations of terrestrial Leiopelma (L. archeyi from Tapu and Tokatea, Coromandel; L. hamiltoni from Stephens Island; L. pakeka from Maud Island). Thirteen presumed genetic (allozyme) loci could be consistently scored for the five populations. At 11 loci, no genetic differences were found between the Whareorino frog and the two Coromandel L. archeyi populations. Allelic frequencies differed slightly at two loci. We therefore conclude that the terrestrial Whareorino frog represents a western population of L. archeyi. L. hamiltoni from Stephens Island is genetically closer to L. archeyi than is L. pakeka from Maud Island. The Whareorino L. archeyi population is morphologically similar to Coromandel L. archeyi populations, although multivariate analysis suggests subtle morphological differences, including the relative position of the nostril. Size comparisons between Whareorino and three Coromandel sites (Moehau, Tapu, Tokatea) show there were more larger frogs (35–38 mm snout‐vent length) at Whareorino and Tokatea compared with Moehau and Tapu, where maximum snout‐vent lengths were 34 and 36 mm, respectively.     

Habitat distribution and predation on a western population of terrestrial Leiopelma (Anura: Leiopelmatidae) in the northern King Country,New Zealand

Abstract

A new population of terrestrial Leiopel‐matid frog was discovered in the Whareorino Forest, northern King Country, New Zealand, in 1991. Searches were carried out from June 1991 to December 1993 to determine the species present and to document variation in external morphology, habitat, and local distribution. These confirmed that a terrestrial frog resembling L. archeyi is present in the area, as well as Hochstetter's frog Leiopelma hochstetteri and the introduced Australian hylid frog Litoria aurea.

In Whareorino Forest, the terrestrial Leiopelma was mostly above 500 m altitude and L. hochstetteri above 350 m. The terrestrial Leiopelma occupies sites under rocks and logs in forest. It also occurs in vegetation, such as crown fern Blechnum discolor, tree fern Cyathea smithii, hook grass Uncinia uncinata, and rice grass Microlaena avenacea. Egg clusters of this frog were found in crown fern and tree fern, as well as under stones.

The terrestrial Leiopelma is susceptible to predation by Litoria aurea and rats. This is the first documented evidence of predation on Leiopelma on the New Zealand mainland. The future of this small remnant Leiopelma population is uncertain, and further investigation of the impact of anuran and mammalian predators is needed.     

The adhesive devices in larvae of Lepidoptera (Insecta, Pterygota)

Adhesion to smooth surfaces by means of thin fluid lipid film was studied on living larvae of 71 species of Lepidoptera by a simple ”light reflection method”. The method made it possible to localize exactly the sites of adhesion and to estimate roughly the film thickness, within a certain range. Furthermore, it revealed the general presence of mobile lipid on the entire insect surface. The observations on living larvae were complemented by comparative structural studies of the adhesive parts with light and scanning electron microscopes on preserved specimens of 161 species. Specialized adhesive devices were found in great diversity on larval legs and prolegs, especially in larvae living in the open air on their food plants. Two main surface types of adhesive cuticle were found: (1) cuticle with a flexible smooth surface and (2) cuticle with very numerous small projections (microtrichia) with spatulate and recurved apices. Both the functional implications of the adhesive cuticular structure and the role of the adhesive fluid as well as the evolution of the adhesive devices are discussed. The adhesive effect is due to ”capillary” or meniscus forces. Accepted: 29 July 1999     

A PCR-based method that permits specific detection of Paenibacillus larvae subsp. larvae, the cause of American Foulbrood of honey bees, at the subspecies level

AIMS: A reliable procedure for the identification of Paenibacillus larvae subsp. larvae, the causal agent of American Foulbrood disease of honey bees (Apis mellifera L.) based on the polymerase chain reaction (PCR) and subspecies - specific primers is described. METHODS AND RESULTS: By using ERIC-PCR, an amplicon of ca 970 bp was found among P. l. larvae strains but not in other closely related species. Based on the nucleotide sequence data of this amplicon, we designed the pair of oligonucleotides KAT 1 and KAT 2, which were assayed as primers in a PCR reaction. A PCR amplicon of the expected size ca 550 bp was only found in P. l. larvae strains. CONCLUSIONS: This PCR assay provides a specific detection for P. l. larvae. SIGNIFICANCE AND IMPACT OF THE STUDY: The developed PCR assay is highly specific because can differentiate Paenibacillus larvae subsp. larvae from the closely related Paenibacillus larvae subsp. pulvifaciens. The technique can be directly used to detect presence or absence of P. l. larvae spores in honey bee brood samples and contaminated honeys.     

Influence of egg size on embryos and larvae of Fundulus heteroclitus (L.)

Egg size for Fundulus heteroclitus (L.) populations is concordant with the distribution of the two F . heteroclitus subspecies, i.e. F. h. heteroclitus eggs are considerably larger than F. h. macrolepidotus eggs. The influence of egg size on survival of embryos during incubation and survival and growth of newly-hatched larvae was estimated for four populations representing both subspecies along the Atlantic coast of the United States and in Delaware Bay. Survival of embryos was determined for incubation periods of 14, 21 and 28 days. Greatest differences in survival were detected following the longest incubation period where less than 50 per cent of the smaller F. h. macrolepidotus eggs survived while little or no mortality was detected among the larger F. h. heteroclitus eggs . Influence of egg size on larval survival was also greatest among those larvae hatched after 28 days where F. h. macrolepidotus larvae survived without food, for an average of 6 days, while F. h. heteroclitus larvae survived 11–12 days. F. h. heteroclitus larvae were significantly larger at hatching than F. h. macrolepidotus larvae. Larval growth rates were the same (0.4 mm day⁻¹) in both subspecies. As a result, size differences at hatching were still maintained after 42 days of growth. The differences in egg size along with other morphological and reproductive characteristics of F. heteroclitus populations probably represent genetically based adaptations to environmental conditions, of which the length of the spawning season is one of the major components stimulating the coevolution of these traits.     

Dytiscus latissimus and D. circumcinctus (Coleoptera,Dytiscidae) larvae as predators on three case-making caddis larvae

The predacious behaviour of Dytiscus circumcinctus and D. latissimus larvae was studied experimentally. When offered different prey simultaneously, D. latissimus larvae preferred cased caddis larvae relative to mayfly nymphs and isopods, whereas in D. circumcinctus the preference order was reversed. Notonectid nymphs and tadpoles were consumed in higher numbers by D. circumcinctus than by D. latissimus larvae. D. circumcinctus larvae and instar III larvae of D. latissimus most frequently captured caddis larvae through the case wall, whereas the instar I and II larvae of D. latissimus normally attacked the thorax of the exposed larva from above the front opening of the case. Limnephilus borealis, L. nigriceps and L. rhombicus caddis larvae differed in case structure, and they were all successfully captured by D. latissimus and D. circumcinctus instar II and III larvae. Neither capture success nor ingestion efficiency varied significantly between the two Dytiscus species or between different prey species. Instar II and III D. circumcinctus larvae had shorter reaction times than those of D. latissimus. The larger L. borealis and L. rhombicus larvae were preferred by the two last Dytiscus larval instars, and the handling time of these two prey was longer than that of L. nigriceps larvae.     

Morphological and behavioural development of halibut, Hippoglossus hippoglossus (L.) larvae

Live yolk-sac halibut, Hippoglossus hippoglossus (L.) larvae from rearing experiments at Austevoll Aquaculture Station, Norway, were examined from hatching to past first feeding for developmental morphology and behaviour. The findings include development of the respiratory and circulatory organs, eye pigmentation, mouth formation, organs of the digestive system and the process of yolk absorption, as well as swimming speed and activity levels.
A stomodeum is not present at hatching although drinking is possible through a pair of branchial pits which gradually develop into the operculum and gill basket. The mouth normally opens slowly, the gape being restricted by a transverse septum until bones are formed. The amount of time spent swimming varies from less than 15% of the observation period during the first 2 weeks after hatching to between 70 and 100% around the seventh week after hatching, when individual differences become more apparent. Larvae generally react with a burst of swimming when two come into contact. Speed and duration of swimming seems to be correlated with development of eye pigment, heart size and fin formation. The yolk-sac period is divided into four stages.     

Giant pelagic larvae of Phyllodocidae (Polychaeta,Annelida)

The microscopic anatomy of giant pelagic larvae of Phyllodocidae was studied using routine histological, SEM, and TEM techniques. The larvae consist of two distinct regions: a large spherical trochophore measuring up to 2 mm in diameter and a posterior, long (up to 10 mm length), narrow rudiment of the adult body with up to 120 segments. The larvae have an unusual mixture of larval and adult features, including a very complex, well-developed brain and ganglia in the ventral nerve cord, and only a single pair of protonephridia located in the hyposphere of the trochophore. A muscular pharynx is not developed. The intestinal wall, especially in the trochophore region, consists of endodermal cells containing considerable nutritive material in the form of yolk-like globular inclusions. The digestive tract of all larvae was empty. The position of the frontal sensory organ and the prototroch, the structure of the parapodia and setae, and the three pairs of tentacular cirri dictate inclusion of the larvae in the family Phyllodocidae. The relatively enormous size and unusual pattern of development of the adult body may be adaptations for a long pelagic life and rapid settlement of the species, which inhabits slopes of islands and underwater mounts located far apart. J. Morphol. 238:93–107, 1998. © 1998 Wiley-Liss, Inc.     

Drinking rate, uptake of bacteria and microalgae in turbot larvae

The drinking rate of turbot larvae increased from 14 to 120 nl larva^-1 h^-1 from day 2 to 11 after hatching, which gave a slightly increased specific drinking rate (calculated per biomass) from day 2 to 7 (0·8–1·9 nl μg carbon^-1 h^-1. The clearance rate of both algae and bacteria was 10–100 times higher than the drinking rate, which indicated that the larvae had an active uptake of both algae and bacteria. On day 2 and 4 after hatching highest clearance rate was observed for Tetraselmis sp. On day 6 about the same clearance rate was observed for bacteria, Isochrysis galbana and Tetraselmis sp. Until day 4 the turbot larvae had a higher ingestion rate of Tetraselmis sp. than of I. galbana , whereas on day 6 the rates were similar (28–41 ng carbon larvae^-1 h^-1). The assimilation efficiency was somewhat higher for I. galbana than for Tetraselmis sp., and on day 6 the assimilated algae constituted 1·5 and 0·9% of the larval biomass for I. galbana and Tetraselmis sp., respectively.     

Host plant utilization by butterfly larvae in the Andaman and Nicobar Islands (Indian Ocean)

The larval food plants of the butterflies of the Andaman and Nicobar islands have not been studied, although the butterfly fauna per se is fairly well known. For the first time we report the food plants of the larvae of 120 species of butterflies from these islands on the basis of laboratory rearing and field studies. This information is essential for the formulation of management programmes for butterfly conservation on these islands which are known to harbour critical swallowtail and (possibly) danaine faunas.     

The effects of temperature and prey density on the development rates and growth of damselfly larvae (Odonata: Zygoptera)

Abstract. 1. The effects of prey density and temperature on the feeding and development rates of several late instars of the larvae of three common European damselflies ( Lestes sponsa (Hansemann), Coenagrion puella (L.) and Ischnura elegans van der Linden) were investigated in a laboratory experiment.
2. Functional responses were used to estimate maximum feeding rates. Maximum feeding rates were compared between species, instars and temperatures by expressing prey consumption in terms of prey biomass consumed per unit predator biomass. Lestes was capable of feeding at almost twice the rate of either Coenagrion or Ischnura.
3. Higher feeding rates led to faster development rates and there was an interaction between species and temperature. With the exception of those with very low feeding rates, larvae maintained at higher temperatures, but similar feeding rates, developed faster. Under similar conditions of temperature and feeding rate, Lestes larvae developed faster than larvae of either Coenagrion or Ischnura.
4. Faster development rates at similar rates of food intake were achieved at the cost of reduced size-increases between instars.
5. The differences in the responses of the three species are discussed in the light of their respective life histories, and with reference to a recent model of population regulation in damselflies.     

A simple, sterile food source for rearing the larvae of Lucilia sericata (Diptera: Calliphoridae)

arvae of the sheep blowfly Lucilia sericata (Meigen) were reared on various mixtures of puréed liver, agar, brewer's yeast and trypsin, in order to develop a simple, sterile, tissue-based diet. Growth and survival rates of larvae reared on a sterile 1:1 mixture of puréed liver with 3% Bacto agar equalled or exceeded those of larvae reared on raw liver. The addition of yeast and/or trypsin to the medium was of no additional benefit. This sterile, homogenous, tissue-based substrate offers a simple, convenient, inexpensive growth medium for rearing larvae for maggot therapy, and for testing the effects of various chemicals and dietary constituents on necrophagous insect larvae. It may be useful, therefore, in studies of myiasis, forensic entomology, and toxicology. This rearing medium also has the advantage that it can be stored for many months at room temperature without progressive decomposition or offensive odour.     

Effects of development, temperature and salinity on metabolism in eggs and yolk-sac larvae of milkfish, Chanos chanos (Forsskål)

Oxygen uptake rates and yolk-inclusive dry weiGhts were measured during the egg and yolk-sac larval stages of milkfish, Chanos chanos (Forsskal). Oxygen uptake by eggs and yolk-sac larvae was measured to assess the effects of four salinities (20,25,30,35 ppt) at 28°C. The effects of three temperatures (23,28,33°C) on oxygen uptake by yolk-sac larvae were determined at a salinity of 35 ppt. Dry weights were measured throughout embryonic development at 28°C and the yolk-sac stage at 23.28 and 33°C.
Oxygen uptake rates of eggs increased more than fivefold during embryogenesis (0.07±0.03 to 0.40 ± 03 μl O₂ egg ⁻¹ h ⁻¹;blastula to prehatch stage). Larval oxygen uptake did not change with age but was affected by rearing temperature (0.33 ± 0.08, 0.44 ± 0.07 and 0.63 ± 0.13 μl O₂ larva ⁻¹ h⁻¹ at 23, 28 and 33°C, respectively; Q₁₀= 1.93). Acute temperature changes from 28 to 33°C caused significant increases in oxygen uptake by embryos (Q ₁₀= 1.69–3.58) and yolk-sac larvae (Q ₁₀=2.55). Salinity did not affect metabolic rates.
Dry weight of eggs incubated at 28°C decreased 13% from fertilization to hatching. Incubation temperatures from 23–33°C did not affect dry weights at hatching. Rearing temperatures significantly affected the rate of larval yolk absorption (Q ₁₀= 2.25).     

Effect of temperature on fatty acid composition and development of unfed Siberian sturgeon (A. baerii) larvae

The fatty acid metabolism in fish is influenced by various factors, including fish species, water temperature, water environment and diet supply. The aim of present work is to investigate the fatty acid composition of yolk‐stage Siberian sturgeon larvae reared at three different temperatures. Fertilized Siberian sturgeon eggs were transferred to the Lodi Aquaculture Research Center of the University of Milan, divided in three aquaria, each containing three incubators and incubated at 16°C. After hatching the temperature was switched to 16, 19 and 22°C. Larvae sampling was performed at the end of yolk sac reabsorption. No feed was dispensed during the trial. Eggs and larvae were weighed and fatty acid profile was determined by GC‐FID analysis after lipid extraction by chloroform/methanol mixture and fatty acid transesterification by methanolic hydrogen chloride. The fertilized eggs had a weight of 23.27 mg and a lipid content of 2.67 mg/egg. At hatching, the weight was 12.2 (0.17 SD) mg and lipid content 1.9 (0.6 SD) mg/larva. At the end of the trial, larvae mean weight was 33.6 (3.6 SD), 34.7 (1.8 SD) and 36.9 (1.1 SD) mg, while lipid content was 2.0 (0.3 SD), 2.1 (0.3 SD) and 2.0 (0.2 SD) mg for larvae reared at 16, 19 and 22°C respectively, without statistically significant difference. Larvae subjected to the highest water temperature showed a faster yolk‐sac absorption. No differences were found across temperatures regarding survival rates and regarding ontogenic development. The fatty acid composition of larvae was affected by the temperature. Larvae reared at 16°C had the lowest amount of saturated fatty acids, mainly due to a lower palmitic acid content, that was offset by a higher level of linolenic and linoleic acid, if compared with larvae reared at 19°C and 22°C. The study suggests that at a lower temperature sturgeon spare unsaturated fatty acid consuming preferably saturated fatty acids, increasing our knowledge of the fatty acid metabolism in this species.     

Distribution and onshore migration behaviour of burbot larvae in Lake Constance, Germany

Larval abundance of burbot Lota lota in the pelagic zone of Lake Constance, Germany, peaked during April and remained at a constant but lower level until mid‐June. From the end of May onwards a significant diel vertical migration pattern was observed: while burbot larvae were distributed above as well as below the thermocline in the daytime with the highest percentages of abundance at water depths of 30 and 50 m (below the thermocline) on 5 and 25 June, they were concentrated from 2 to 15 m (above the thermocline) during the night. This migration pattern coincided with a significant decline in the food items preferred by burbot at this life stage, cyclopoid copepods. This phase of diel vertical migration lasted for c . 4 weeks until the burbot larvae suddenly disappeared from the pelagic zone. This indicated a short phase, lasting only a few days, of larval settlement into profundal benthic habitats. This was supported by a significant, abrupt decrease in the absolute growth rates of the larvae at day 56 after hatching. The day–night pattern of vertical migration observed could be assumed to be a transition phase from a pelagic to a benthic lifestyle, as reported for juveniles of several other, marine gadoid species.     

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.