Objective: To explore the relationship between extracellular enzymes activity and virulence of Candida glabrata clinical isolates based on the infection model of Galleria mellonella larvae. Methods: Using experimental research methods, 71 strains of non-repetitive Candida glabrata were collected from Qinghai Provincial People's Hospital from June 2021 to January 2022. Bovine serum protein agar medium, egg yolk agar medium, sheep blood agar medium, Tween-80 agar medium and triglyceride agar medium were used to detect the aspartyl protease activity, phospholipase activity, hemolysis activity, esterase activity and lipase activity of Candida glabrata. Median lethal concentration (LC₅₀) was calculated by using 1.25×10⁸ CFU/ml,2.50×10⁸ CFU/ml,3.75×10⁸ CFU/ml,5.00×10⁸ CFU/ml suspension of Candida glabrata ATCC2001 to infect Galleria mellonella larvae. Histopathological and etiological analysis was performed to determine whether the infection model was successfully established. The clinical isolates of Candida glabrata were configured to infect Galleria mellonella larvae with LC₅₀ concentration to detect the pathogenicity of Galleria mellonella larvae.Spearman test or Pearson test were used to analyze the correlation between the extracellular enzyme activity of Candida glabrata clinical isolates and the pathogenicity of Galleria mellonella larvae. Results: 71 strains of Candida glabrata isolated clinically were detected to have low hemolytic activity after 2 days of culture. Aspartyl protease was detected after 4 days of culture, among which 7 strains (9.86%), 19 strains (26.76%) and 45 strains (63.38%) showed low, medium and high aspartyl protease activity. After 7 days of culture, 71 strains did not detect phospholipase, esterase and lipase activities. Candida glabrata on Galleria mellonella larvae of LC₅₀=2.5×10⁸ CFU/ml Fungal spore were found in the intestinal tissue pathological section of Galleria mellonella larvae in the experimental group, and Candida glabrata was identified by the microbial Mass Spectrometry after culture, while no fungi were found in the pathological section and culture of the control group. Spearman test shows that, there was a linear positive correlation between aspartyl protease activity and the survival rate of Galleria mellonella larvae (r = 0.73, P<0.01), the difference was statistically significant.Pearson test shows that, there was no significant linear relationship between hemolytic activity and survival rate of Galleria mellonella larvae (r = 0.16, P = 0.34), the difference was not statistically significant. Conclusion: The clinical isolates of Candida glabrata in this study had aspartyl protease activity and low hemolytic activity, but no phospholipase, esterase and lipase activity. The activity of aspartyl aspartyl protease of Candida glabrata was positively correlated with the pathogenicity of Galleria mellonella larvae.
Animals ; Sheep ; Larva/microbiology* ; Virulence ; Candida glabrata ; Agar ; Moths/microbiology* ; Esterases ; Aspartic Acid Proteases ; Lipase

The observation statistics suggested that the haemolymph melanization speed of larvae became fast and the growth inhibition of Escherichia coli was strong as the quantities of feeding on mulberry leaves increased. The RT-PCR result showed that the mRNA expressions of melanin biosynthesis enzyme BmTan, BmPo-1, BmYellow-f and BmDdc were high in the haemolyph of 5 L 3 d larvae. The qPCR analysis showed Bmtan, Bmddc, Bmyellow, Bmebony and Bmblack, especially Bmddc expression were significantly higher in black disease larvae than in normal larvae. Compared with control, Ddc inhibitors drastically inhibited the lipopolysaccharide-induced haemolymph melanization. In addition, the content of Dopa and Dopamine markedly rose after E. coli injection. These indicated that haemolymph melanization was linked to immune defenses and Bmddc may play a role in melanization response of haemolymph immune in silkworm.
Animals ; Bombyx ; enzymology ; genetics ; microbiology ; Escherichia coli ; Genes, Insect ; Hemolymph ; chemistry ; Larva ; Melanins ; biosynthesis

OBJECTIVEAssessment of the bacterium L. sphaericus as a dual-action candidate for biological control of mosquito-borne diseases and bioremediation of toxic metals.

METHODSLarvae of the mosquito, C. quinquefasciatus, were first evaluated for metal tolerance and then exposed to 5 ppm cadmium, chromium, arsenic, and lead in assays together with seven strains of L. sphaericus. A probit regression analysis was used to estimate the LC(50) of Cd, Cr, As, and Pb to C. quinquefasciatus. An analysis of covariance and multifactorial ANOVA examined the metal biosorption and larvicidal properties of the seven strains of L. sphaericus.

RESULTSWe found that L. sphaericus adsorbed the toxic metal ions and was toxic against mosquito larvae. The L. sphaericus strain III(3)7 resulted in a larvae mortality of over 80% for all the tested metals. This strain also exhibited the capacity to adsorb 76% of arsenic, 32% of lead, 25% of chromium, and 7% of cadmium.

CONCLUSIONThis study found combined metal adsorption and larval toxicity associated with three strains of L. sphaericus [III(3)7, OT4b.31, and CBAM5]. This suggests that a combination of these strains shows strong dual potential for biological control of mosquitos in heavy metal-contaminated areas and remediate the heavy metal contamination as well.

Animals ; Bacillaceae ; physiology ; Culicidae ; microbiology ; Host-Pathogen Interactions ; Insect Vectors ; Larva ; microbiology ; Metals, Heavy ; metabolism ; toxicity ; Water Pollutants, Chemical ; metabolism ; toxicity

To clear the effect of the wound to the growth of the larva of the host to the Ophiocordyceps sinensis, the wounds of same severity at the same position were made artificially to the larva and which were artificial fed at the same environment and condition. The results indicated that, over the winter, the survival rate of the wounded of the infection larva was lower than that of the healthy larva, but the weight had no significant difference between the wounded and the healthy larva. The survival rate of the wounded of the no infection larva was lower than that of the healthy larva, but except with black skin, the wounded larva with offwhite and dusty red had no influence on the variety of the weight. In summery, wound had no advantage to the survival rate, but had no influence to the weight. The result had provided theoretical basis to the reforming of the system of the artificial culture O. sinensis.
Animals ; Body Weight ; Breeding ; methods ; Hypocreales ; growth & development ; Larva ; Moths ; growth & development ; microbiology

A total of 9,281 larval chigger mites were collected from small mammals captured at Hwaseong-gun, Gyeonggi-do (Province) (2,754 mites from 30 small mammals), Asan city, Chungcheongnam-do (3,358 mites from 48 mammals), and Jangseong-gun, Jeollanam-do (3,169 for 62 mammals) from April-November 2009 in the Republic of Korea (= Korea) and were identified to species. Leptotrombidium pallidum was the predominant species in Hwaseong (95.8%) and Asan (61.2%), while Leptotrombidium scutellare was the predominant species collected from Jangseong (80.1%). Overall, larval chigger mite indices decreased from April (27.3) to June (4.9), then increased in September (95.2) and to a high level in November (169.3). These data suggest that L. pallidum and L. scutellare are the primary vectors of scrub typhus throughout their range in Korea. While other species of larval chigger mites were also collected with some implications in the transmission of Orientia tsutsugamushi, they only accounted for 11.2% of all larval chigger mites collected from small mammals.
Animals ; Arachnid Vectors ; Larva/*microbiology ; Orientia tsutsugamushi/*isolation & purification ; Republic of Korea ; Rodentia ; Scrub Typhus/*microbiology ; Trombiculidae/*classification/*microbiology

To address the role of Pr1 gene in the process of Hirsutella sinensis infecting Hepialus gonggaensis, differential expression of subtilisin-like protease gene was detected. In the present study, Pr1 gene analogues from H. sinensis were obtained by PCR strategy using specific primers designed from conserved regions of Pr1 gene reported in the GenBank. Then we detected the changes in the expression of Pr1 gene before and after infecting H. gonggaensis using real-time quantitative PCR. We obtained the partial sequence of Pr1 gene with the length of 535 bp (GenBank accession: KC009680). Real-time PCR results showed that the expression level of Pr1 gene was significantly different among 8 samples (P < 0.01). Pr1 gene showed the obvious higher expression level (2-3 folds) after infecting the H. gonggaensis, suggesting that the Pr1 gene may play an important role in the process of H. sinensis infecting H. gonggaensis. The present study paves a way for further identification on infectivity assessment of H. sinensis.
Amino Acid Sequence ; Animals ; Hypocreales ; genetics ; metabolism ; pathogenicity ; Larva ; Lepidoptera ; microbiology ; Real-Time Polymerase Chain Reaction ; Subtilisin ; genetics ; metabolism

OBJECTIVETo investigate the toxicity of indigenous Bacillus thuringiensis (B. thuringiensis)isolates from Malang City for controlling Aedes aegypti (Ae. aegypti) larvae.

METHODSSoil samples were taken from Purwantoro and Sawojajar sub-districts. Bacterial isolation was performed using B. thuringiensis selective media. Phenotypic characteristics of the isolates were obtained with the simple matching method. The growth and prevalence of spores were determined by the Total Plate Count method, and toxicity tests were also performed on the third instar larval stage of Ae. aegypti. The percentage of larval mortality was analysed using probit regression. The LC50 was analysed by ANOVA, and the Tukey HSD interval was 95%.

RESULTSAmong the 33 selected bacterial isolates, six were obtained (PWR4-31, PWR4-32, SWJ4-2b, SWJ4-4b, SWJ-4k and SWJ5-1) that had a similar phenotype to reference B. thuringiensis. Based on the dendrogram, all of the bacterial isolates were 71% similar. Three isolates that had a higher prevalence of reference B. thuringiensis were PWR4-32, SWJ4-4b and SW5-1, of which the spore prevalence was 52.44%, 23.59%, 34.46%, respectively. These three indigenous isolates from Malang City successfully killed Ae. aegypti larvae. The PWR4-32 isolates were the most effective at killing the larvae.

CONCLUSIONSSix indigenous B. thuringiensis isolates among the 33 bacterial isolates found in the Sawojajar and Purwantoro sub-districts were toxic to the third instar larvae of Ae. aegypti. The PWR4-32 isolates were identical to the reference B. thuringiensis and had 88% phenotype similarity. The PWR4-32 isolates had the highest spore prevalence (52.44%), and the early stationary phase occurred at 36 h. The PWR4-32 isolates were the most effective at killing Ae. aegypti larvae (LC50-72 h=2.3×10(8) cells/mL).

Aedes ; microbiology ; Animals ; Bacillus thuringiensis ; isolation & purification ; physiology ; Biological Control Agents ; Indonesia ; Insecticides ; Larva ; microbiology ; Lethal Dose 50 ; Mosquito Control

Strongyloides stercoralis is a soil transmitted intestinal nematode that is endemic in the tropical and subtropical regions. In most individuals who are infected, chronic, usually asymptomatic, gastrointestinal infection persists. But, in immunocompromized hosts or in patients receiving immunosuppressive therapy, autoinfection of S. stercoralis may result in the dissemination of larvae, leading to fatal hyperinfection and increased rate of complications. We report a case of hyperinfective strongyloidiasis with bacterial meningitis in a patient receiving steroid therapy. Strongyloidiasis was diagnosed by the presence of filariform larvae of S. stercoralis in the bronchoalveolar lavage cytology and upper gastrointestinal endoscopic biopsy specimen. Her clinical symptoms had progressively aggravated and developed bacterial meningitis during treatment. She died despite aggressive antibiotic and antihelminthic therapy.
Adrenal Insufficiency/drug therapy ; Aged ; Animals ; Bronchoalveolar Lavage Fluid/parasitology ; Endoscopy, Gastrointestinal ; Enterococcus faecium/isolation & purification ; Female ; Humans ; Immunocompromised Host ; Intestinal Mucosa/pathology ; Larva/physiology ; Magnetic Resonance Imaging ; Meningitis, Bacterial/complications/*diagnosis/microbiology ; Steroids/adverse effects/therapeutic use ; Strongyloides stercoralis/growth & development/isolation & purification ; Strongyloidiasis/complications/*diagnosis/parasitology

OBJECTIVETo clarify the relevance of amino acid content in cultivated Cordyceps and cultivation materials.

METHODThe content of amino acid was determined with L-8800 amino acid analyzer, and the relevance of amino acid content was analyzed with SPSS.

RESULT AND CONCLUSIONExcept mycelium of the C. sinensis or the blood-lymph of the larva, the cultivated Cordyceps and the main relevant cultivation materials had detected to contain all kinds of amino acids. Except among the mycelium, the blood-lymph of the larva, the part of the larva or of the stroma of cultivated Cordyceps, there was distinct relevance of amino acid contents in cultivated Cordyceps and the cultivation materials (P<0.01).

Amino Acids ; analysis ; metabolism ; Animals ; Cordyceps ; chemistry ; metabolism ; Larva ; chemistry ; microbiology ; Moths ; chemistry ; microbiology ; Mycelium ; chemistry ; metabolism

OBJECTIVETo study the biological character of Hepialus introduced from Yunnan province.

METHODH. jianchuanensis introduced from Yunnan was bred artificially to observe its biological character.

RESULTThe pupa duration was about 40 d, pupa emergence lasted for one day, female to male ratio was about 1.2:1, the mating peak was 19:30-20:30, the adults duration was about 25 d, the average life time of female was 5-6 d and that of male 6-7 d, the average number of the producing egg was about 300, the average weight of the 100 eggs was 10.33 mg. Lasting period of the larva is 350 d and the infancy larva showed the character of the polyphagy and cold endurance.

CONCLUSIONResults indicate that there are some differences in the whole duration, weight of the 100 eggs and the number of the eggs left in the abdomen between H. jianchuanensis and others bred at the Kangding base.

Animals ; Arachnid Vectors ; microbiology ; physiology ; China ; Cold Temperature ; Feeding Behavior ; physiology ; Female ; Larva ; anatomy & histology ; physiology ; Male ; Moths ; anatomy & histology ; physiology ; Oviposition ; physiology ; Ovum ; physiology ; Pupa ; physiology