Novel Myxobolus and Ellipsomyxa species (Cnidaria: Myxozoa) parasiting Brachyplatystoma rousseauxii (Siluriformes: Pimelodidae) in the Amazon basin,Brazil

We describe two novel myxosporean parasites from Brachyplatystoma rousseauxii, an economically important freshwater catfish from the Amazon basin, Brazil. Myxobolus tapajosi n. sp., was found in the gill filaments of 23.5% of 17 fish, with myxospores round to oval in frontal view and biconvex in lateral view: length 15 (13.5–17) μm and width 10.7 (9.6–11.4) μm; polar capsules equal, length 5.8 (4.6–7.1) μm and width 3 (2.3–3.8) μm containing polar tubules with 6–7 turns. Ellipsomyxa amazonensis n. sp. myxospores were found floating freely or inside plasmodia in the gall bladder of 23.5% of fish. The myxospores were ellipsoidal with rounded extremities: length 12.8 (12.3–13.6) μm and width 7.6 (6.7–8.7) μm; with two equal, slightly pyriform polar capsules, length 3.8 (3.8–4.0) μm and width 3.1 (2.5–3.4) μm, containing polar tubules with 2–3 turns. We combined spore morphometry, small-subunit ribosomal DNA data, specific host, and phylogenetic analyses, to identify both of these parasites as new myxozoan species. Maximum likelihood and Bayesian analyses showed that Myxobolus tapajosi n. sp. clustered in a basal branch in a subclade of parasites from exclusively South American pimelodid fishes. Ellipsomyxa amazonensis n. sp. clustered within the marine Ellipsomyxa lineage, but we suspect that although the parasite was collected in freshwater, its hosts perform a large migration throughout the Amazon basin and may have become infected from a brackish/marine polychaete host during the estuary phase of its life.     

Wing development and parthenogenesis induced in progenies of kinoprene-treated gynoparae of Aphis fabae and Myzus persicae

Presumptive gynoparae of Aphis fabae and Myzus persicae were exposed to various levels of kinoprene (Zoecon's ZR 777) by being placed as 4th-instar alatiform larvae on bean or radish seedlings that had been sprayed with different concentrations of kinoprene in an acetone-tween-water emulsion. Larvae exposed to the highest (0.1%) concentration tested developed into adults 1 to 2 days sooner than those on control plants. The adults on the treated plants had variously deformed wings, reduced sclerotization (and pigmentation in the case of M. persicae) and other apteriform features. On reaching adulthood the affected aphids settled to feed and started to larviposit some days earlier than the control aphids. After two weeks as adults, treated gynoparae of M. persicae produced more larvae than the 7 to 9 typically deposited by control gynoparae under the short-day and cool temperature conditions employed in these tests.Whereas most or all of the larvae produced by the control gynoparae developed into oviparae (apterous, egg-laying, sexual females), gynoparae exposed to 0.1% kinoprene-treated plants predominantly produced alatiform viviparous offspring. If the latter were allowed to develop on untreated plants they deposited a few oviparous larvae. Alatiform virginoparae of M. persicae (from the same holocyclic strain that produced the gynoparae) also responded to kinoprene by developing wing deformities and by producing alatiform offspring. In contrast, alatiform virginoparae from an androcyclic strain of M. persicae, although developing wing deformities, produced only apterous progeny.The stimulation by kinoprene of wing development and parthenogenesis in the progeny of treated gynoparae is discussed in the light of our present knowledge of these aspects of aphid polymorphism.     

Observations on egg production in the monogenean Entobdella soleae

Egg production in the monogenean Entobdella soleae increases as adult parasites grow. Medium-sized adults (anterior hamulus length 550–600 μm; total body length approx. 5 mm) produced about 30 eggs per day at 12°C. Parasites with anterior hamuli exceeding 650 μm in length (total body length about 6 mm) may produce more than 60 eggs per day. In smaller adults, eggs tend to spend longer in the ootype after assembly of the shell and the ootype remains empty for longer periods of time. The time taken to assemble a single egg is relatively constant (4–6 min) in adults of all sizes and there is evidence of a small but significant increase in egg size as parasites increase in size. Some eggs were retained in the uterus for as long as 168 min but others spent less than 5 s in the uterus, indicating that the uterus is no more than a passageway for these eggs to the outside world. The egg cell (or zygote) precedes the vitelline cells as they progress along the ovo-vitelline duct to the ootype.     

Molecular data and phylogenetic analysis of Myxobolus pseudonobilis n. sp. infecting the gill filaments of silver carp,Hypophthalmichthys molitrix Valenciennes, 1844

In the present study, we combined morphological and phylogenetic methods to characterize Myxobolus pseudonobilis n. sp. infecting Hypophthalmichthys molitrix Valenciennes, 1844 from Chongqing, China. The morphology and molecular characteristics of M. pseudonobilis n. sp. were distinct from those of other previously described Myxobolus species. Mature myxospores were ovoid in frontal view with spore dimensions of 10.0 ± 0.4 (9.3–10.9) μm in length and 8.5 ± 0.2 (7.9–9.0) μm in width. Two polar capsules occupying approximately half of the myxospore length were unequal in size. The larger polar capsule containing 6 to 7 filament coils measured 5.2 ± 0.3 (4.5–5.8) μm in length and 3.6 ± 0.2 (3.2–3.9) μm in width, while the smaller capsule with 4 to 5 filament coils measured 3.9 ± 0.3 (3.0–4.4) μm in length and 2.5 ± 0.3 (2.1–3.6) μm in width. The comparison of molecular characteristics demonstrated similarities and genetic distances of 18S rDNA sequences of 95.19% - 98.20% and 1.82% - 5.46%, respectively, between M. pseudonobilis n. sp. and its morphologically similar species, and secondary structures were also distinctly different. Moreover, phylogenetic analysis showed that M. pseudonobilis n. sp. was clustered with other myxobolids possessing spores with a blunt anterior end and branched independently. In addition, the morphology of myxosporeans as an important indicator was discussed.     

Tip growth of <Emphasis Type="Italic">Neurospora crassa</Emphasis> upon resource shortage: Disturbances of the coordination of elongation,branching, and septation

The characteristics of elongation, branching, septation, and nuclear morphology in hyphal tips (of ~400 μm in length) of the mycelial fungus Neurospora crassa isolated from the mycelium and cultivated for several hours have been investigated using intracellular fluorescent markers. The newly formed branches had the following characteristic features: (1) the predefined orientation was conserved, whereas the diameter decreased (from 10–20 to 6.5 ± 0.4 μm), as did the elongation rate (from 24 ± 1 to 6.7 ± 0.5 μm/min); (2) a disturbed branching pattern with abnormally large internodal distances (up to 1471 μm) and developmental arrest of part of the buds of lateral branches; and (3) a conserved septation pattern and a relatively constant length of hyphal segments (68 ± 2 μm). The size of the nucleus-free zone at the tip (5–33 μm) and the distance between the first septum and the growth point (210 ± 15 μm) in the daughter branches of the isolated fragments were almost the same as in hyphae connected to the mycelium, whereas the average distance between the growth point and the first lateral branch (492 ± 127 μm) and the variability of this parameter were higher in the isolated fragments. The morphology of the nuclei and the size of the nucleus-free zone near the growth point did not differ from those reported for normal vegetative hyphae of N. crassa. The experimental model developed may be used for the elucidation of details of molecular genetic mechanisms that underlie the regulation of interactions between the intracellular structures that provide tip growth of the hyphae in N. crassa.     

Juvenile hormone and photoperiodically controlled polymorphism in Aphis fabae: Prenatal effects on presumptive oviparae

All three naturally occurring juvenile hormones (JH's) were shown to have effects on the parthenogenetic/gamic polymorphism of Aphis fabae; they mimicked long day conditions by inducing parthenogenetic forms. When topically applied to fourth instar gynoparae, JH caused the appearance of oviparous/viviparous intermediate morphs in the progeny. JH induced both wing development and embryogenesis in embryonic, presumptive oviparae. Embryogenesis was induced by lower doses of JH. Adult, embryo-containing alatae produced by treatment with high JH doses were capable of flight, and whilst reluctant to reproduce, their few viable progeny were oviparae. They did, however, differ from normal gynoparae in size, occasional presence of scent plaques on the metathoracic tibiae, numbers of secondary rhinaria on the antennae and morphogenetic response to postnatal rearing in long day conditions. The presumptive, oviparous embryos most sensitive to JH treatment were shown to be ca 323 μm in length, close to the stage where their germaria differentiate as parthenogenetic or gamic. Similar effects were observed in the progeny of JH-treated, teneral adult gynoparae but there was no effect on the morph of progeny of long day, alate virginoparae. The JH's differed in potency in the order JH I > JH II > JH III. The treatment of fourth instar gynoparae also induced a terminal batch of apparently normal viviparous progeny in a number of aphids. This result was obtained even at JH doses below threshold for the appearance of oviparous/viviparous intermorphs.     

Benzofuran-isatin hybrids tethered via different length alkyl linkers and their in vitro anti-mycobacterial activities

A series of novel benzofuran-isatin hybrids 6a–m tethered through different length alkyl linkers propylene, butylene, pentylene and hexylene were designed, synthesized and evaluated for their in vitro anti-mycobacterial activities against both drug-susceptible and multi-drug resistant (MDR) Mycobacterium tuberculosis (MTB) and cytotoxicity towards VERO cells. All hybrids with acceptable cytotoxicity in VERO cells (CC₅₀: 64 to >1024 μg/mL) also exhibited considerable anti-mycobacterial activities against both drug-susceptible and MDR-MTB strains with MIC in a range of 0.125–4 μg/mL. The SAR indicated that the length of the linker played a pivotal role on the activity, and the longer linker could enhance the activity. The most active hybrid 6d (MIC: 0.125 and 0.125 μg/mL) was comparable to or better than rifampicin (MIC: 0.5 μg/mL) and isoniazid (MIC: 0.06 μg/mL) against MTB H₃₇Rv, and was ≥256 folds more potent than rifampicin (MIC: 64 μg/mL) and isoniazid (MIC: >128 μg/mL) against MDR-MTB strain, but was less active than TAM16 (MIC: <0.06 μg/mL against the tested two strains). The hybrid 6d also showed low cytotoxicity towards VERO cell (CC₅₀: 128 μg/mL), but it was inferior to TAM16 in metabolic stability and in vivo pharmacokinetic profiles.     

Evaluation of a briquet formulation of Bacillus thuringiensis var. israelensis (H‐14) against aedes spp. and culex spp. larvae in dambos in Kenya

Bacillus thuringiensis Berliner var. israelensis serotype H‐14 (Bti) in briquet formulation (Bactimos) was tested in a field trial against ground‐pool breeding mosquitoes in a dambo located in a Rift Valley fever virus‐enzootic/epizootic area in central Kenya. Bactimos (10% Bti; 7000 AA International Toxic Units/mg) was tested for 30 days in 3 separate treatment areas at the rate of 1 briquet/9 m2, later increased to 1 briquet/1.5 m2 on day 13 postflood; 1 briquet/9 m2; and 1 briquet(4.6 m2). An estimate of the daily survival rate of larvae at different periods during the study revealed that mosquitoes in the area treated with 1 briquet/9 m2 had a significantly lower (64%) survival rate than those in the control site (92%) against Aedes spp. and was not significantly different from the site treated with 1 briquet/4.6 m². Mosquitoes in the site treated with I briquet/1.5 m² on day 13 postflood had a much‐reduced survival rate (25%;) when compared to those in the control site (67%) during the 3rd Culex spp. generation (22‐ 26 days postflood). This five‐fold increase above standard label dosage still failed to prevent Culex spp. emergence. There was no significant difference in survival rates between the control and any of the three treatment sites during the first or second Culex generation (9–20 days postflood). Emerged adults collected as pupae from control and treatment sites indicated that Aedes mcintoshi Huang and Culex antennatus (Becker) were the most predominant Aedes and Culex species.     

Differential pulse polarographic determination of plasma menadione

A differential pulse polarographic assay for plasma vitamin K₃ (menadione) has been developed. Details of the assay are (i) lipid-soluble material is extracted from plasma into ether by the method of Bjornsson et al. [(1978) Thromb. Haemostas.2, 466–473]; (ii) ether is evaporated under nitrogen and the residue is dissolved in the supporting electrolyte, methanol: 0.2 m borate buffer (9:1), pH 6.8; (iii) current height is measured at ?0.32 V vs SCE on the differential pulse polarogram. The lower sensitivity limit of this technique after addition of standard vitamin K₃ to plasma is 0.3 μm; the calibration curve is linear from 0.6 through 10 μm. Two patients treated with a single dose of menadiol sodium diphosphate, 20 mg/M² i.m., achieved measurable plasma vitamin K₃ levels at 0.5 to 1.0 h ranging between 0.5 (0.08 μg/ml) and 2 μm (0.3 μg/ml).     

Morphological,ultrastructural and phylogenetic analyses of Myxobolus hilarii n. sp. (Myxozoa,Myxosporea), a renal parasite of farmed Brycon hilarii in Brazil

Myxobolus hilarii n. sp. was described, based on morphology, histology, ultrastructure and 18S rDNA sequencing, infecting the kidney of Brycon hilarii (Valenciennes 1850) (Characiformes: Bryconidae) taken from fish farms in the state of São Paulo, Brazil. Thirteen specimens of B. hilarii were examined and 100% had round, white plasmodia in the kidney. The mature myxospores were rounded, measuring 11.5 ± 0.8 (9.8–13.4) μm in length, 11.0 ± 0.7 (9.7–12.4) μm in width and 7.6 ± 1.0 (6.7–9.0) μm in thickness. Polar capsules were elongated and of equal size, with 6.5 ± 0.4 (6.0–7.2) μm in length and 4.0 ± 0.2 (3.6–5.3) μm in width and their polar filaments had 5 to 7 coils. Histological analysis revealed plasmodial development in the renal tubules, causing compression and deformation of adjacent tissues and destruction of renal tubule cells. Ultrastructural analysis showed direct contact between the plasmodial wall and the host tissue and asynchronous plasmodial development. The phylogenetic analysis of South American myxobolids, based on 18S rDNA sequencing, showed the myxosporeans grouping into two main clades. M. hilarii n. sp. appears as sister species of Myxobolus piraputangae.     

Potential of Lecanicillium spp. for management of insects, nematodes and plant diseases

Fungi in the genus Lecanicillium (formerly classified as the single species Verticillium lecanii) are important pathogens of insects and some have been developed as commercial biopesticides. Some isolates are also active against phytoparasitic nematodes or fungi. Lecanicillium spp. use both mechanical forces and hydrolytic enzymes to directly penetrate the insect integument and the cell wall of the fungal plant pathogen. In addition to mycoparasitism of the plant pathogen, the mode of action is linked to colonization of host plant tissues, triggering an induced systemic resistance. Recently it was demonstrated that development of Lecanicillium hybrids through protoplast fusion may result in strains that inherit parental attributes, thereby allowing development of hybrid strains with broader host range and other increased benefits, such as increased viability. Such hybrids have demonstrated increased virulence against aphids, whiteflies and the soybean cyst nematode. Three naturally occurring species of Lecanicillium, L. attenuatum, L. longisporum, and an isolate that could not be linked to any presently described species based on rDNA sequences have been shown to have potential to control aphids as well as suppress the growth and spore production of Sphaerotheca fuliginea, the causal agent of cucumber powdery mildew. These results suggest that strains of Lecanicillium spp. may have potential for development as a single microbial control agent effective against several plant diseases, pest insects and plant parasitic nematodes due to its antagonistic, parasitic and disease resistance inducing characteristics. However, to our knowledge, no Lecanicillium spp. have been developed for control of phytopathogens or phytoparasitic nematodes.     

Residual force enhancement and force depression in human single muscle fibres

Residual force depression (rFD) and residual force enhancement (rFE) are intrinsic contractile properties of muscle. rFD is characterized as a decrease in steady-state isometric force following active shortening compared with a purely isometric contraction at the same muscle length and level of activation. By contrast, isometric force is increased following active lengthening compared to a reference isometric contraction at the same muscle length and level of activation; this is termed rFE. To date, there have been no investigations of rFD and rFE in human muscle fibres, therefore the purpose of this study was to determine whether rFD and rFE occur at the single muscle fibre level in humans. rFD and rFE were investigated in maximally activated single muscle fibres biopsied from the vastus lateralis of healthy adults. To induce rFD, fibres were activated and shortened from an average sarcomere length (SL) of 3.2–2.6 μm. Reference isometric contractions were performed at an average SL of 2.6 μm. To induce rFE, fibres were actively lengthened from an average SL of 2.6–3.2 μm and a reference isometric contraction was performed at an average SL of 3.2 μm. Isometric steady-state force was lower following active shortening (p < 0.05), and higher following active lengthening (p < 0.05), as compared to the reference isometric contractions. We demonstrated rFD and rFE in human single fibres which is consistent with previous animal models. The non-responder phenomenon often reported in rFE studies involving voluntary contractions at the whole human level was not observed at the single fibre level.     

Reproductive biology of the antarctic opisthobranch Philine gibba Strebel

Monthly samples (August 1974 to February 1976) and cultured material have been used to describe ovotestis maturation, spawning, and larval development of the Antarctic opisthobranch Philine gibba Strebel. Ovotestis maturation took one year and mature individuals (mean shell length 12 ± 1.4 mm) spawned a single egg mass in April–May containing 975–4760 eggs, 379 μm in diameter.Development was non-pelagic but an active veliger was observed within the egg capsule. Juveniles (mean shell length 375 ± 55 μm), which hatched from August to October, resembled adults except for the presence of eye spots and an exposed shell. The mantle enclosed the shell by February when shell length was 890 ± 30 μm.     

<Emphasis Type="Italic">Myxobolus imparfinis</Emphasis> n. sp. (Myxozoa: Myxosporea), a new gill parasite of <Emphasis Type="Italic">Imparfinis mirini</Emphasis> Haseman (Siluriformes: Heptapteridae) in Brazil

A new species of myxozoan, Myxobolus imparfinis n. sp. is described based on material from the gills of Imparfinis mirini (Haseman) (Heptapteridae). Mature myxospores are round, measuring 7.1–8.4 (7.9 ± 0.3) μm in length, 4.5–6.2 (5.5 ± 0.5) μm in width and 3.1–4.2 (3.7 ± 0.3) μm in thickness. The polar capsules are of unequal size, the larger polar capsule measuring 3.4–4.5 (3.9 ± 0.3) μm in length and 1.4–2.0 (1.7 ± 0.1) μm in width and the smaller capsule measuring 3.1–3.8 (3.4 ± 0.2) μm in length and 1.2–1.8 (1.5 ± 0.2) μm in width. The polar filament presents 6–7 coils. Spores had a prevalence of infection of 75% (6/8). In histological analyses we detected the development site of spores in primary filaments, in afferent branchial artery, thus classifying the type of infection to the filamental type and vascular subtype. The phylogenetic analyses of a dataset including species Myxobolus Bütschli, 1882 and Henneguya Thélohan, 1892 from South America recovered M. imparfinis n. sp. as a sister species of Myxobolus flavus Carriero, Adriano, Silva, Ceccarelli & Maia, 2013. To our knowledge, this is the first record of a myxozoan species parasitising I. mirini.     

Alteration in properties of thymidylate synthetase from pyrimethamine-resistant Plasmodium chabaudi

Alteration in properties of thymidylate synthetase from pyrimethamine-resistant smodium chabaudi. International Journal for Parasitology16: 483–490. Thymidylate synthetase from cloned strains of pyrimethamine-sensitive and resistant P. chubaudi were partially purified and characterized. The enzyme from both strains have equal mol. wt of 120,000 as estimated by Sephadex G-200 column chromatography. The enzyme from drug-sensitive parasites has an optimum pH of 6.5–7.5 and is stable at pH 4–11 while that from drug-resistant strain has an pH optimum of 7.0–8.0 and is stable at pH 5–10. The K_m for methylenetetrahydrofolate are 206 ± 6 and 495 ± 5 μm for the enzyme from drug-resistant and sensitive parasites, respectively. The K_m for dUMP of the enzyme from drug-resistant and sensitive parasites are 42 ± 1 and 49 ± 6 μm, respectively. Inhibition of the enzyme from both strains by FdUMP are competitive with dUMP; however,the K_is for the enzyme from drug-resistant strain (0.043 ± 0.005 μm) is less than that from drug-sensitive strain (0.11 ± 0.007 μm) by a factor of 2.5. The K_ii for methotrexate with respect to methylenetetrahydrofolate of the enzyme from drug-resistant parasites (58 ± 3 μm) is 3 times larger than that from drug-sensitive parasites (17 ± 1 μm).     

Taxonomy,phylogeny and host-parasite interaction of two novel Myxobolus species infecting Brycon orthotaenia from the São Francisco River,Brazil

Two new Myxobolus species were described infecting Brycon orthotaenia from the São Francisco River, in the state of Minas Gerais, Brazil. From a total of 39 B. orthotaenia collected, two specimens (5.1%) exhibited infection of the ovary and 12 specimens (30.8%) displayed infection of the liver. The plasmodia of both Myxobolus species were white and spherical measuring around 1 mm in length. The plasmodium found in the ovary showed mature myxospores, which were oval shaped from the frontal view and measured 9.2–11.0 (9.8 ± 0.4) μm in length, 5.9–6.9 (6.5 ± 0.3) μm in width and 4.6–5 (4.9 ± 0.1) μm in diameter. The two polar capsules were the same size and measured 3.9–6.2 (4.7 ± 0.5) μm in length and 1.8–2.4 (2.1 ± 0.2) μm in width. The polar tubules had 9 coils. The plasmodium found in the liver showed mature myxospores which were ellipsoidal in shape from the frontal view and measured 10.0–11.4 (10.7 ± 0.5) μm in length, 7.3–8.6 (8.1 ± 0.4) μm in width and 5.3–7.0 (6.8 ± 0.4) μm in diameter. The two polar capsules were the same size and measured 4.2–5.4 (4.9 ± 0.3) μm in length and 1.9–2.9 (2.7 ± 0.3) μm in width. The polar tubules had 8 coils. Ultrastructural analysis revealed an asynchronous sporogenesis process, with young developmental myxospore stages more often found in the periphery of the plasmodium and mature myxospores in the centre of the plasmodium. The plasmodial wall was formed by a single membrane which was not surrounded by a layer of host tissue. A thick layer of fibrous material was found in the peripheral ectoplasm close to the plasmodial wall of the plasmodium found in the ovary. Phylogenetic analysis based on the small-subunit ribosomal DNA – ssrDNA sequences and using the closest myxozoan sequences to each one of the species studied here based on previous GenBank data and Henneguya/Myxobolus/Thelohanellus species parasitizing fish from South American, revealed that the new species are grouped in a subclade together with other Myxobolus species parasitizing bryconid hosts.     

A new species of Eimeria Schneider, 1875 (Apicomplexa: Eimeriidae) from the dusky rice rat Melanomys caliginosus (Tomes) (Mammalia: Rodentia) in Costa Rica

We collected faecal samples from 24 dusky rice rats, Melanomys caliginosus (Tomes) (Rodentia: Cricetidae: Sigmodontinae), in a Biological Reserve in Costa Rica, and found three (12.5%) to be infected with a species of Eimeria Schneider, 1875, which we describe here as new. Sporulated oöcysts of Eimeria caliginosa n. sp. are almost spheroidal and measure 16–21 × 17–20 (mean 19.6 × 18.2) μm; micropyle, oöcyst residuum and polar granule are absent. Sporocysts are ovoidal, 9–13 × 6–8 (mean 11.2 × 6.7) μm, with small Stieda and sub-Stieda bodies present, but a para-Stieda body is absent; the sporocyst residuum is a compact mass of c.11–15 granules, c.5 μm wide. Sporozoites are crescent-shaped, 5–8 × 2–3 (mean 6.8 × 2.4) μm. This is the third species of Eimeria described from the genus Melanomys Thomas.     

LIFE CYCLE,SIZE REDUCTION PATTERNS,AND ULTRASTRUCTURE OF THE PENNATE PLANKTONIC DIATOM PSEUDO‐NITZSCHIA DELICATISSIMA (BACILLARIOPHYCEAE)1

Pseudo‐nitzschia delicatissima (Cleve) Heiden is a very common pennate planktonic diatom found in temperate marine waters, where it is often responsible for blooms. Recently, three distinct internal transcribed spacer types have been recorded during a P. delicatissima bloom in the Gulf of Naples (Mediterranean Sea, Italy), which suggests the existence of cryptic diversity. We carried out mating experiments with clonal strains belonging to the most abundant internal transcribed spacer type. Pseudo‐nitzschia delicatissima is heterothallic and produces two functional anisogametes per gametangium. The elongated auxospore possesses a transverse and a longitudinal perizonium. The sexual phase was observed to occur over a wide size spectrum, spanning 19–80 μm and corresponding to almost the whole range of cell length observed for P. delicatissima. We also investigated cell morphology, valve ultrastructure and morphometry of parental, F1‐generation strains, and the progeny of crosses between parental and F1 strains. Although ultrastructural features match those described for P. delicatissima, variability in cell shape was recorded in the largest cells of the F1 generation as well as in valves with an abnormal arrangement of poroids. As many other diatoms, P. delicatissima undergoes size reduction over its life cycle, and cells of different size showed differences in growth rates and the amount of size reduction per cell cycle. Cells between 60 and 30 μm in length showed the fastest growth and the slowest rates of size reduction per generation. In culture, P. delicatissima cells can decrease to 8 μm in length; however, such small cells (≤30 μm) are not recorded in the sea, and this raises interesting questions about the factors that control their survival in the natural environment.     

Host instar preference and functional response of Aphidius transcaspicus, a parasitoid of mealy aphids (Hyalopterus species)

Aphidius transcaspicus Telenga (Hymenoptera: Braconidae: Aphidiinae) has been selected for use as a biological control agent against Hyalopterus pruni Geoffroy (Hemiptera: Aphididae) in prune orchards in California. In this study we focus on the host instar preference and functional response of A. transcaspicus. Host instar preference was determined using laboratory arenas by allowing female parasitoids to attack aphids on field-collected leaves of Prunus domestica L. supporting a natural range of H. pruni instars. A relative preference index indicated that A. transcaspicus was 1.4 times more likely to attack intermediate instars (2–4) than 1st or adult instars. The functional response of A. transcaspicus to H. pruni density was determined on enclosed P. domestica shoots in a prune orchard. The functional response to host density was linear (Type I) at the lowest aphid densities, and the asymptotic number of aphids attacked at higher aphid densities was 64.3?±?6.7 aphids day^?1.     

Rhythm of honeydew excretion by the tea aphid and its attraction to various natural enemies

The rhythm of honeydew excretion by the tea aphid Toxoptera aurantii (Boyer) and its attraction to following 9 species (or subspecies) of beneficial insects, Aphidius sp., Chrysopa sinica Tjeder, Chrysopa septempunctata Wesmael, Sphaerophoria menthastri L., Coccinella septempunctata L., Leis axyridis (Pallas) ab. bimaculata Hemmelmann, L. axyridis (Pallas) ab. conspicua Faldermaenn, L. axyridis (Pallas) var. spectabilis Faldermaenn and L. axyridis (Pallas) var. novemdecimpunctata Faldermaenn, were investigated. Forty-five wingless virginoparae nymphs, reproduced by the same wingless virginogenia adult within 1 h, were introduced onto tea seedlings with one aphid per seedling. The honeydews excreted from different instars of nymphs and adults were collected under 21°C, 85% RH, 3500 lx and 12 L:12 D photoperiod. It took 32.4 d ±5.8 d for the tested tea aphids to complete the development of their nymph and adult stages, during which 325.6±35.8 droplets (ca. 41.98 μl ±6.14 μl and 45.34 mg ± 8.76 mg) of honeydews were secreted. During the 1st and 4th instars, there was a logistic regression relationship between the amount of honeydews excreted and the time (days). The honeydew secretion during the first two instars was less than that from the later instars. Adult aphids survived for 22.0 d ± 0.0 d, and excreted 176.31 ± 22.38 droplets (ca. 30.38 μl ± 5.32 μl) of honeydews in a rate of 1 drop per ca. 30–50 min for 5–8 h with a pause for 2–5 h before next secretion series. A batch of forty-five virginoparae female adults, reproduced by the same virginoparae female adult within 1 h, were introduced onto the tea seedlings (one aphid/seedling) under 13–21°, 85% RH, 3500 lx and 12 L:12 D photoperiod. Temperature showed a significant effect on the amounts of honeydews excreted within the range of 13–21°. Honeydews excreted by the aphids significantly increased the searching and retention time of the tested 9 species of natural enemies in a positive dose-response fashion. The searching times of Aphidius sp. and S. menthastri were the longest and the shortest, respectively, among all the 9 species, while the searching and retention time of L. axyridis (Pallas) var. spectabilis was the longest among the four varieties of L. axyridis. Tea aphid oneydew is considered as an important contact kairomone for the tested natural enemies.