Post-endoscopic submucosal dissection electrocoagulation syndrome: a clinical overview.

INTRODUCTION: Post-endoscopic submucosal dissection electrocoagulation syndrome (PEECS) is a rare complication following endoscopic submucosal dissection (ESD). However, many aspects of PEECS are still controversial and there is a lack of up-to-date review. The aim of this article is to provide a comprehensive review on PEECS. AREAS COVERED: The English-language literature was searched for articles on PEECS, with a focus on its pathogenesis, definition, and diagnosis, incidence rate, risk factors, prevention, management, and prognosis. EXPERT OPINION: Many aspects of PEECS are still controversial such as etiology, diagnostic criteria, prevention and management. The recent studies tend to agree on the diagnostic criteria for PEECS, defined as localized abdominal tenderness and fever or inflammatory response without delayed perforation. Most patients with PEECS have a favorable prognosis with conservative treatment and more high-quality studies are needed in the controversial aspects mentioned above.

Distribution and pathogenicity of Beauveria bassiana in soil with earthworm action and feeding.

Earthworm action and feeding have an important impact on a variety of microorganisms in the soil. However, the effects of the earthworm on Beauveria bassiana, a common entomopathogenic fungus in the biological control of pests, have been little studied. In this study, the epigeic earthworm species Eisenia fetida (Savigny) was selected to evaluate its impact on B. bassiana TST05 including its distribution in soil and its pathogenicity to target insects. By testing B. bassiana TST05 distribution, biomass in soil, viable spore germination rate, and pathogenicity to insect larvae after passing through the earthworm gut, the results showed that the activity and feeding of E. fetida promoted the B. bassiana TST05 diffusing downwards in the soil, while decreasing active fungal spores. After passing through the earthworm gut and excretion, the living B. bassiana individuals still had activity and pathogenicity to insects. The germination rate of the viable fungal spores was 15.09% and the infection rate to the insect larvae of Atrijuglans hetaohei Yang reached 62.35%, 80.95% and 100% after infection at 7 d, 10 d, and 14 d, respectively. The results showed that action and feeding of earthworms promoted the distribution of B. bassiana TST05 in soil, but decreased B. bassiana viable spores. This study is important for understanding the interaction between earthworms and B. bassiana in soil and for guiding the scientific application of B. bassiana in the biological control of pests.

Effect of earthworm Eisenia fetida epidermal mucus on the vitality and pathogenicity of Beauveria bassiana.

Beauveria bassiana is one of the most widely studied and used entomopathogenic fungus as biopesticide. In the biological control of pests, B. bassiana will persist in the soil after application, and will inevitably contact with earthworms, especially the epigeic earthworm species. So, what are the effects of earthworm and its epidermal mucus on the activity of B. bassiana? We employed the epigeic earthworm Eisenia fetida, B. bassiana TST05 strain, and the insect Atrijuglans hetaohei mature larvae to study the impact of earthworm epidermal mucus on the vitality and pathogenicity of B. bassiana to insect. Methods included scanning electron microscope observation, detection of spore germination, fungal extracellular enzyme activity, and infection testing to A. hetaohei. The results showed that the B. bassiana spores may attach to the cuticle of E. fetida but they could be covered by the epidermal mucus and became rough and shrunken. After treatment with the epidermal mucus, the spore germination and extracellular enzymes of B. bassiana was significantly inhibited. Inoculation of A. hetaohei larvae with a mixture of B. bassiana and mucus showed that the mucus could reduce the pathogenicity of B. bassiana to the insect, resulting in a slower disease course and lower mortality. It was concluded that the epidermal mucus of the earthworm E. fetida can inhibit the activity of B. bassiana, as well as the infectivity and pathogenicity of fungus to target insects. However, after treatment with epidermal mucus the surviving B. bassiana still had certain infectivity to insects. This is of great significance for the application of B. bassiana in biological control of pests.

Curcumin intervention for cognitive function in different types of people: A systematic review and meta-analysis.

Curcumin is a polyphenolic natural compound with diverse and attractive biological activities, which may prevent or ameliorate pathological processes underlying age-related cognitive decline, dementia, or mood disorders. However, clinical trials and animal studies have yielded conflicting conclusions regarding its effectiveness for cognition in different individuals. The aim of this review is to meta-analytically assess the effectiveness of curcumin for cognitive function in different types of people. A preliminary search on PubMed, Embase, Web of Science, ClinicalTrials.gov, Cochrane Library, Chinese National Knowledge Infrastructure, and Wanfang Data and China Biology Medicine disc was performed to identify randomized controlled trials investigating the effect of curcumin on cognition. Six clinical trials with a total of 289 subjects met inclusion criteria for this review. We used a random-effects model to calculate the pooled standardized difference of means (SMD). For older adults who received curcumin, scores on measures of cognitive function (SMD = 0.33, 95% confidence interval [CI] [0.05, 0.62]; p = 0.02), occurrence of adverse events (odds ratio [OR] = 5.59, 95% CI [0.96, 36.80]; p = 0.05), and measures of depression (SMD = -0.29, 95% CI [0.64, 0.05]; p = 0.09) indicated significant memory improvement. In patients with Alzheimer's disease (AD), scores in measures of cognition status (SMD = -0.90, 95% CI [1.48, -0.32]; p = 0.002) indicated that there was a trend for treated subjects to do worse than placebo-treated subjects on the Mini-Mental State Examination. The occurrence of adverse events (OR = 0.87, 95% CI [0.10, 7.51]; p = 0.90) was similar to those who received placebo. Due to insufficient data, it was impossible to provide a narrative account of only the outcomes for schizophrenia. Curcumin appears to be more effective in improving cognitive function in the elderly than in improving symptoms of AD and schizophrenia. Curcumin is also safe and tolerated among these individuals. Because of the small number of studies available, a funnel plot or sensitivity analysis was not possible. Further high-quality trials with larger sample sizes or bioavailability-improved curcumin formulations may be considered for reliable assessment.

Streptococcus himalayensis sp. nov., isolated from the respiratory tract of Marmota himalayana.

Five strains of Gram-positive-staining, catalase-negative, coccus-shaped, chain-forming organisms isolated separately from the respiratory tracts of five Marmota himalayana animals in the Qinghai-Tibet Plateau of China were subjected to phenotypic and molecular taxonomic analyses. Comparative analysis of the 16S rRNA gene indicated that these singular organisms represent a new member of the genus Streptococcus, being phylogenetically closest to Streptococcus marmotae DSM 101995T (98.4 % similarity). The groEL, sodA and rpoB sequence analysis showed interspecies similarity values between HTS2T and Streptococcus. marmotae DSM 101995T, its closest phylogenetic relative based on 16S rRNA gene sequences, of 98.2, 78.8 and 93.7 %, respectively. A whole-genome phylogenetic tree built from 82 core genes of genomes from 16 species of the genus Streptococcus validated that HTS2T forms a distinct subline and exhibits specific phylogenetic affinity with S. marmotae. In silico DNA-DNA hybridization of HTS2T showed an estimated DNA reassociation value of 40.5 % with Streptococcus. marmotae DSM 101995T. On the basis of their phenotypic characteristics and phylogenetic findings, it is proposed that the five isolates be classified as representatives of a novel species of the genus Streptococcus, Streptococcus himalayensis sp. nov. The type strain is HTS2T (=DSM 101997T=CGMCC 1.15533T). The genome of Streptococcus himalayensis sp. nov. strain HTS2T contains 2195 genes with a size of 2 275 471 bp and a mean DNA G+C content of 41.3 mol%.

Antennal sensilla and their nerve innervation in first-instar nymphs of Porphyrophora sophorae Arch. (Hemiptera: Coccomorpha: Margarodidae).

Porphyrophora (Hemiptera: Coccomorpha: Margarodidae) is a genus of soil-inhabiting scale insects. The antennal sensilla and their innervation in the first-instar nymphs of Porphyrophora sophorae were studied using light microscopy and scanning and transmission electron microscopy to understand the function of these sensilla and determine the sensillar innervation feature on these small antennae. The results show that the six-segmented antennae of these nymphs have 20-23 sensilla which can be morphologically classified into seven types, for example, one Böhm's bristle (Bb), one campaniform sensillum (Ca), one Johnston's organ (Jo), 13-16 aporous sensilla trichodea (St), two coeloconic sensilla (Co), one straight multiporous peg (Mp1), and one curvy multiporous peg (Mp2). According to their function, these sensilla can be categorized into three categories: mechanoreceptors, that is, Bb, Ca, Jo, and St; thermo/hygroreceptors, that is, Co only; and chemoreceptors, that is, Mp1 and Mp2. The dendrites that innervate the Mp1, Mp2, and Co sensilla combine to form a large nerve tract (NT1) in the antennal lumen. Because NT1 extends through and out of the antenna, the somata of these neurons are present in the lymph cavity of the insect's head. The dendrites that innervate the mechanoreceptors form another nerve tract (NT2). The somata of these neurons are located inside the scape and pedicel. J. Morphol. 277:1631-1647, 2016. © 2016 Wiley Periodicals, Inc.

Streptococcusmarmotae sp. nov., isolated from the respiratory tract of Marmota himalayana.

Five strains of a Gram-stain-positive, catalase-negative, α-haemolytic, coccus-shaped chain-forming organism were isolated separately from the lower respiratory tracts of five animals of Marmota himalayana in the endemic area of plague, the Qinghai-Tibet Plateau, China. Based on their morphological characteristics, biochemical features and molecular phylogenetic studies, the strains were placed as representing a new member of the genus Streptococcus. Comparative 16S rRNA gene sequence studies indicated that strain HTS5T shared 96.5, 96.2 and 96.0 % similarity with Streptococcus gallinaceus CCUG 42692T, Streptococcus parasanguinis ATCC 15912T and Streptococcus suis ATCC 43765T, respectively. Sequence analysis of its rpoB and sodA genes showed that strain HTS5T was most closely related to Streptococcus cuniculi CCUG 65085T with 9.2 and 10.9 % interspecies divergence, respectively. The whole genome phylogenetic tree based on 339 core genes of 65 Streptococcus genomes confirmed that HTS5T belongs to a distinct lineage that is well separated from recognized species of the genus Streptococcus. In silico DNA-DNA hybridization using 65 available genomes from GenBank showed that HTS5T displayed less than 70 % DNA-DNA relatedness with the other 65 species of the genus Streptococcus deposited in the GenBank database. The genome of strain HTS5T (2 322 791 bp) contained 2377 genes and had a G+C content of 41.6 mol%. Therefore, the five strains are considered to represent a novel species of the genus Streptococcus for which the name Streptococcusmarmotae sp. nov. is proposed. The type strain is HTS5T (=DSM 101995T=CGMCC 1.15534T).

Streptococcus halotolerans sp. nov. isolated from the respiratory tract of Marmota himalayana in Qinghai-Tibet Plateau of China.

Two Gramstaining-positive, catalase-negative, α-hemolytic, coccus-shaped organisms were isolated separately from the respiratory tracts of two Marmota himalayana animals from the Qinghai-Tibet Plateau, PR China. Morphological, biological, biochemical, and molecular genetic studies were performed on these two isolates (HTS9T and HTS12). Their biochemical characteristics, such as acid production from different sugars and enzymatic activities, indicated that they represented a member of the genus Streptococcus. They are most closely related to Streptococcus thoraltensis CIP 105518T based on sequence analysis of their 16S rRNA, groEL, sodA and rpoB genes, with similarities of 97.6, 89.9, 92.6 and 91.1 % the four genes respectively. The whole genome phylogenetic tree reconstructed using 372 core genes from 65 genomes of members of the genus Streptococcus validates that HTS9T forms a distinct subline and exhibits specific phylogenetic affinity with S. thoraltensis. In silico DNA-DNA hybridization of HTS9T showed a DNA reassociation value of 32.1 %, closest to that of S. thoraltensis CIP 105518T. Based on their phenotypic characteristics and in particular the phylogenetic findings (DNA-DNA hybridization, three phylogenetic trees built from the partial 16S rRNA/housekeeping genes, and from 372 core genes of 65 genomes of members of the genus Streptococcus), we propose with confidence that strains HTS9T and HTS12 should be classified as representing a novel species of the genus Streptococcus, Streptococcus halotolerans sp. nov. The type strain is HTS9T (=DSM 101996T=CGMCC1.15532T). Genome analysis of Streptococcus halotolerans sp. nov. shows that its genome is 1 823 556 bp long with a DNA G+C content of 39.9 mol% and contains 2068 genes.

The structure and morphogenic changes of antennae of Matsucoccus matsumurae (Hemiptera: Coccoidea: Matsucoccidae) in different instars.

To better understand the functioning and morphogenic changes of the antennae of Matsucoccus matsumurae (Kuwana) in different instars, the antennae are examined using light microscopy, scanning and transmission electron microscopy. The results show that the antennae of M. matsumurae display three different styles in morphology and sensillar distribution in different instars. The antennae of first instar nymphs are relatively simple, including one campaniform sensillum (Ca), four smooth aporous trichoid sensilla (SAt), two intersegmental sensilla (Ins), two coeloconic sensilla (Co), three multiporous pegs (Mp) and four uniporous pegs (Up). The antennae of adult females and third instar male nymphs both possess similar antennae, and exhibit seven types of sensilla. Adult female antennae have in total 82-108 sensilla, including 9-16 Böhm's bristle (Bb), 3-7 Ca, 50-75 SAt, 0-3 Ins, 3-10 Co, 8 Mp and 5 Up, whereas third instar male nymph antennae possess approximately 62-79 sensilla. Adult male antennae are the most developed, possessing 259-312 sensilla, including 7-15 Bb, 2-5 Ca, 7-11 grooved aporous trichoid sensilla, 4-9 SAt, 0-3 Ins, 2-7 Co, 23-29 knobbed seta sensilla, 179-230 multiporous trichoid sensilla and 8 Mp. Based on these results, the main functions and morphogenic changes of antennae M. matsumurae in different instars are discussed.

Pathogenicity of three entomopathogenic fungi to Matsucoccus matsumurae.

Matsucoccus matsumurae (Kuwana) (Hemiptera: Coccoidea: Matsucoccidae) is an invasive alien species and a destructive pest of two native Chinese pines, Pinus tabulaeformis Carr. and P. massoniana Lamb., throughout the eastern regions of China. The pathogenicity of three entomopathogenic fungi, Lecanicillium lecanii strain V3.4504 and V3.4505, Fusarium incarnatum-equiseti strain HEB01 and Lecanicillium fungicola strain HEB02, against M. matsumurae was tested in four instars, to evaluate their potential as a biological control agent. The results showed that the four strains caused disease and death of the scale insect, among which the L. lecanii strains V3.4504 and V3.4505 displayed stronger virulence than the F. incarnatum-equiseti strains HEB01 and L. fungicola strain HEB02 to M. matsumurae in the 2nd-instar nymphs and the adult females. Furthermore, L. lecanii V3.4505 was most virulent to M. matsumurae. The adult females and the male 3rd-instar nymphs of M. matsumurae were susceptible to L. lecanii V3.4505; the adult females were more susceptible at LT50 = 1.96 than the 3rd-instar nymphs at LT50 = 5.67. The body surface structure, cuticle thickness and wax secretions of M. matsumurae impacted the fungal infection. L. lecanii is a promising biocontrol agent, and newly emerged male 3rd-instar nymphs and adult females are a crucial period of the insect's life cycle for M. matsumurae biocontrol.

Ultra- and microstructure of the female reproductive system of Matsucoccus matsumurae.

The ultra- and microstructure of the female reproductive system of Matsucoccus matsumurae was studied using light microscopy, scanning and transmission electron microscopy. The results revealed that the female reproductive system of M. matsumurae is composed of a pair of ovaries, a common oviduct, a pair of lateral oviducts, a spermatheca and two pairs of accessory glands. Each ovary is composed of approximately 50 telotrophic ovarioles that are devoid of terminal filaments. Each ovariole is subdivided into an apical tropharium, a vitellarium and a short pedicel connected to a lateral oviduct. The tropharium contains 8-10 trophocytes and two early previtellogenic oocytes termed arrested oocytes. The trophocytes degenerate after egg maturation, and the arrested oocytes are capable of further development. The vitellarium contains 3-6 oocytes of different developmental stages: previtellogenesis, vitellogenesis and choriogenesis. The surface of the vitellarium is rough and composed of a pattern of polygonal reticular formations with a center protuberance. The oocyte possesses numerous yolk spheres and lipid droplets, and is surrounded by a mono-layered follicular epithelium that becomes binucleate at the beginning of vitellogenesis. Accessory nuclei are observed in the peripheral ooplasm during vitellogenesis.

The wax glands and wax secretion of Matsucoccus matsumurae at different development stages.

In this paper, the wax secretions and wax glands of Matsucoccus matsumurae (Kuwana) at different instars were investigated using light microscopy, scanning electron microscopy and transmission electron microscopy. The first and second instar nymphs were found to secrete wax filaments via the wax glands located in the atrium of the abdominal spiracles, which have a center open and a series of outer ring pores. The wax gland of the abdominal spiracle possesses a large central wax reservoir and several wax-secreting cells. Third-instar male nymphs secreted long and translucent wax filaments from monolocular, biolocular, trilocular and quadrilocular pores to form twine into cocoons. The adult male secreted long and straight wax filaments in bundles from a group of 18-19 wax-secreting tubular ducts on the abdominal segment VII. Each tube duct contained five or six wax pores. The adult female has dorsal cicatrices distributed in rows, many biolocular tubular ducts and multilocular disc pores with 8-12 loculi secreting wax filaments that form the egg sac, and a rare type wax pores with 10 loculi secreting 10 straight, hollow wax filaments. The ultrastructure and cytological characteristics of the wax glands include wax-secreting cells with a large nucleus, multiple mitochondria and several rough endoplasmic reticulum. The functions of the wax glands and wax secretions are discussed.

The effect of Beauveria brongniartii and its secondary metabolites on the detoxification enzymes of the pine caterpillar, Dendrolimus tabulaeformis.

The mortality of pine caterpillar, Dendrolimus tabulaeformis Tsai et Liu (Lepidoptera: Lasiocampidae), larvae treated with Beauveria brongniartii (Saccardo) Petch (Hypocreales: Clavicipitaceae) conidia and cell-free culture supernatants enriched for the secondary metabolites of the fungus was investigated. In addition, the effects of the treatments on the activities of two insect-related defense response proteins, glutathione S-transferase (GST) and esterase (EST), were measured over time. Bioassays were performed using a range of fungal spore (6 × 105 through 6 × 107 spores/mL) and supernatant extract concentrations (5.5-550 µg/mL). The results showed that the mortalities of D. tabulaeformis larvae were closely related to the concentration of the conidia and the metabolites of B. brongniartii. The differences among the treatments all reached a significant level. The activities of the two detoxifying enzymes, GST and EST, in the larvae increased simultaneously post-treatment. After infection with the conidial suspensions, the highest GST activity appeared at 3 days, and the activities of the caterpillars infected with 6 × 106 spores/mL and 6 × 107 spores/mL were significantly higher than in the control. Using α-naphthyl, the highest activity of EST also appeared at 3 days, and the differences for the three different concentrations were significant. A similar trend of change in the EST activity was observed using ß-naphthyl. After treatment with the secondary metabolite solution, the highest GST activity appeared at 6 hr, and significant differences were found both for the different durations (2, 4, 6, 12, 24, and 48 hr) and in the three concentration groups. When using α-naphthyl, the EST activity peak appeared at 24 hr, and the differences were significant among the durations of 2, 4, 6, 12, 24, and 48 hr. The effect of the concentration of the secondary metabolite solution notably induced the EST activity in the insects, and a similar result was obtained using ß-naphthyl. The data suggest that B. brongniartii produces secondary metabolites that disable the immune mechanisms of D. tabulaeformis, allowing the fungus to overcome and then kill its host. It was concluded that both the conidial suspensions and the metabolites of B. brongniartii were toxic to D. tabulaeformis larvae.

Cellular apoptosis of hemocytes from Dendrolimus tabulaeformis Tsai et Liu larvae induced with the secondary metabolites of Beauveria brongniartii (Sacc.) Petch.

To investigate the effect of the secondary metabolites of entomopathogenic fungus on the hemocyte immunity of host insect, the secondary metabolite complex (SMC) of Beauveriabrongniartii was used in three concentrations (5.5, 55, and 550 µg/mL), and the 4(th) instar larvae of the pine caterpillar Dendrolimustabulaeformis were employed as host insects. The larvae were inoculated with the SMC solutions by injection in bioassays. Apoptosis of the larval hemocytes was observed using fluorescence microscopy (FM), transmission electron microscopy (TEM), and flow cytometry (FCM). The FM results showed that in the treated groups, larval hemocytes exhibited symptoms of early apoptosis at 6 h post-treatment by radiating a non-uniform kelly fluorescence and exhibited symptoms of late apoptosis at 12 h post-treatment by radiating a non-uniform orange fluorescence. Under TEM, the following ultra-structural changes associated with apoptosis of the larval hemocytes were observed in the treated groups: the nuclei were hypertrophied, slight folds were on the nuclear envelope, the chromatin became concentrated, the mitochondrial cristae disappeared or were disorderly, most cells developed blebs, and fibrillar aggregation appeared and accumulated in the cytoplasm. Apoptosis of the larval hemocytes was detected by FCM at 6 h post-treatment; the percentage of early apoptotic cells in the SMC 5.5, 55, and 550 µg/mL treatment groups were 11.93%, 13.10%, and 18.42%, respectively. Late apoptosis first occurred at 12 h post-treatment; the highest rate of apoptosis was 36.54 ± 4.37% at 24 h post-treatment in the SMC 55 µg/mL treatment group. In general, the cellular apoptosis rate was positively correlated with the SMC concentration and the time post-treatment. These results indicate that secondary metabolites of B. brongniartii are able to attack the hemocytes of D. tabulaeformis larvae and induce cellular apoptosis, thereby providing new evidence that secondary metabolites of mycopathogens can act on host immune systems.

[Use of entomopathogenic fungi to degrade wax secreted by Ceroplastes japonicus].

OBJECTIVE: We used entomopathogenic fungi to degrade insect wax. METHODS: We used four fungal strains, Lecanicilliurn lecanii V3. 4504, V3. 4505, Beauveria bassiana FDB01, and Metarhizium anisopliae TSL06. Wax coverings of female adults of Ceroplastes japonicus Green (Insecta: Hemiptera: Coccoidea) were used as the sole carbon source in the mineral medium. RESULTS: All of the 4 strains could grow, reproduce, produce enzymes, and degrade wax. During a 7-day culture, the highest lipase activities of the 4 strains, V3. 4504, V3. 4505, FDB01, and TSL06 were 0.128 +/- 0.017, 0.056 +/- 0.002, 0.124 +/- 0.011, and 0.149 +/- 0.005 U/mL, respectively. The dehydrogenases activities of the 4 strains were 0.075 +/- 0.003, 0.074 +/- 0.003, 0.061 +/- 0.04, and 0. 066 +/- 0. 002 U/mL respectively. The degradation rates of wax by the 4 strains were 18.20 +/- 0.019, 11.00 +/- 0.011, 15.4 +/- 0.017, and 23.10 +/- 0.031%, respectively. CONCLUSION: The 4 strains could depredate wax of C. japonicus.

Morphological and ultrastructural characterization of Carposina sasakii larvae (Lepidoptera: Carposinidae) infected by Beauveria bassiana (Ascomycota: Hypocreales: Clavicipitaceae).

The aim of this study was to better understand the pathogenesis of the entomopathogenic fungus Beauveria bassiana (Balsamo) strain TST05 observed on the peach fruit moth (Carposina sasakii (Matsumura)), an important orchard pest. The morphological and ultrastructural characterization of the mature larvae of C. sasakii infected by B. bassiana was investigated by using light, scanning and transmission electron microscopy. The results of the study show that B. bassiana TST05 infected the host larvae mainly by penetrating the integument. The conidia of the fungus adhere easily to the area around the mouthparts and to the basal area around the acanthae on the thorax and abdomen. Observations of the host's defensive response to the fungal attack indicated that dark spots appeared on the cuticle and that melanization appeared in the hemocoel. After overcoming the host's defense system, the pathogen grew and reproduced primarily in the hemocoel. The infection spread sequentially to the internal tissues, e.g., fat body, muscle, Malpighian tubules, gut and even the silk gland. Ultimately, the larval internal organs and tissues were damaged very extensively. Finally, the fungus emerged through the cuticle of the dead insect and released conidiophores that could act as new pathogens to infect other larvae.

The structure of integument and wax glands of Phenacoccus fraxinus (Hemiptera: Coccoidea: Pseudococcidae).

Using scanning electron microscopy and optical microscopy, we studied the structure of the integument and wax glands of the mealybug, Phenacoccus fraxinus Tang (Hemiptera: Coccoidea: Pseudococcidae). We observed the ultrastructure of four wax pores including trilocular, quinquelocular, and multilocular pores as well as tubular ducts, recording characteristics of their structure, size and distribution. We found that that the integument of the mealybug consists of three main layers-the procuticle, epidermis and basement membrane-and four sub-layers of the procuticle-the epicuticle, exocuticle, endocuticle and formation zone. The wax-secreting gland cells were closely arranged in epidermis. All of them were complex and composed of one central cell and two or more lateral cells. These complex cells possess a large common reservoir for collection and storage. Synthesized by the glandular cells, the wax is excreted outside integument through canals.

Morphological and ultrastructural characterization of the alimentary canal in Japanese wax scale (Ceroplastes japonicus Green).

The morphology and ultrastructure of the alimentary canal in the adult female of the Japanese wax scale, Ceroplastes japonicus Green (Hemiptera: Coccoidea: Coccidae), was investigated using light microscopy, scanning electron microscopy and transmission electron microscopy. The results showed that the foregut was subdivided into a sclerotized pharynx and an oesophagus. A pair of salivary glands attached in the middle of the foregut. The loop-shaped midgut was narrow and longer than the foregut and its inner wall lined with a thick layer of epithelia. The hindgut was divided into a narrower ileum and a broader rectum, with the well-developed filter chamber enclosed in the anterior rectum. Malpighian tubules consisted of two brownish-yellow moniliform tubules with pores, approximately 1 µm in diameter, scattered on the outer surface and many spherical crystals inside the tubules.

Ultrastructural and cytochemical characterization of brown soft scale Coccus hesperidum (Hemiptera: Coccidae) infected by the Lecanicillium lecanii (Ascomycota: Hypocreales).

The ultrastructural and cytochemical characterization of the brown soft scale, Coccus hesperidum L. (Hemiptera: Coccidae) infected by the hyphomycete Lecanicillium lecanii (Zimmermann) Gams & Zare, belonging to the phylum Ascomycota and order Hypocreales, was investigated by light, scanning and transmission electron microscopy. Gold cytochemistry was used to label chitin in the cuticle of the scale insect. The results revealed that the pathogenic fungus, L. lecanii generally infected by penetrating the integument, especially at anus, vulva, spiracles, stigmatic furrow, body margin, and the areas of cuticle with grooves, fissures and rugoses areas. The conidia became attached to the host body surface and germinated into hyphae that established colonies by branching repeatedly. Hyphae penetrated the integument by means of their penetration pegs using mechanical force and extracellular enzymes. During integument penetration, the hyphae extended vertically or parallel along the cuticle. Labeling with the WGA/Ovo-G complex showed disruption of the parallel sheets of chitin and a decrease in the density of the gold particles surrounding the penetrated hyphae. Hyphal invasion also separated the cuticle and epidermis from each other. Once in the haemocoele, blastospores of the fungus infected the haemocytes and internal organs. After some time, the nutritive value of the haemocoele decreased and the insect's internal organs disappeared. The hyphae then produced conidiophores and released them through the cuticle of the scale insect cadaver.

[Effect of multi-generation culture on virulence of Lecanicilliurn lecanii in different media].

OBJECTIVE: The strain No. V3.4504 of Lecanicilliurn lecanii (Zimmermann), an entomopathogenic fungus, was studied on the effect of successive multi-generation culture in seven different media on its colony growth characteristics, extracellular enzyme activities and the virulence against scale insects. METHODS: The strain No. V3.4504 of L. lecanii was original isolated from a natural infected scale insect. The two species of scale insects used were Rhodococcus sariuoni Borchsenius and Ceroplastes japonicus Green. Seven media were used and fungus colony characteristics, growth rate and sporulation, extracellular protease and chitinase activity, and infective effect against the two species of scale insects were conducted. RESULTS: The fungus cultured on PDA medium for successive nine generations showed the most fast in colony growth, the minimal in sporulation, straight decline of extracellular protease and chitinase activity with generation increasing, and the minimal mortality of the scale insects. There was no significant effect to promote virulence of the fungus by increasing peptone into medium. On the media D, E and F, that with the body materials of the two scale insects, although the fungus appeared lower in the colony growth rate, its sporulation was higher upward 8.83 x 10(6) - 9.13 x 10(6) spores/cm2, extracellular protease and chitinase activities averagely reached 2.16 - 2.13 U/g and 1.01 - 1.03 U/g respectively, and the mortalities of the two scale insects were 55% - 58% and 39% - 42% respectively. Cultured three generations in vitro of the two scale insects, the fungus exhibited the highest activities in its protease and chitinase that were 3.08 - 2.92 U/g and 1.45 - 1.42 U/g respectively and the best infection effect against the two scale insects with mortalities of 71.30% and 58.89% respectively. A linear correlation was found between extracellular protease and chitinase activities of the fungus and the mortalities of the scale insects. CONCLUSION: Cultured on PDA medium successive multiple generations made retrogradation of the strain No. V3.4504 of of L. lecanii. It was significant effect on keeping the vigor and higher virulence of the fungus adding the body materials of the scale insects into the medium. The vitro by using live scale insects as medium materials was the best way for the rejuvenation of the entomopathogenic fungus and promoting its virulence.