Degenerative process and cell death in salivary glands of Rhipicephalus sanguineus (Latreille, 1806) (Acari: Ixodidae) semi-engorged female exposed to the acaricide permethrin

Ticks are ectoparasites of great medical and veterinary importance around the world and synthetic chemicals such as permethrin have been used for their control. This study provides a cytochemistry analysis of both degenerative and cell death processes in salivary glands of the brown dog tick Rhipicephalus sanguineus semi-engorged females exposed to 206, 1,031, and 2,062 ppm of permethrin. The results presented herein demonstrate that permethrin is a potent chemical acaricide that would act on the glandular tissue's morphophysiology in this tick species by eliciting severe changes in the acinus shape, intense vacuolation of the acinar cells' cytoplasm, marked glandular tissue disorganization, culminating in an advanced degenerative stage with consequent formation of many apoptotic bodies (cell death). In addition, permethrin induced major changes in the acinar cells' nucleus, such as a change both in its shape and size, chromatin marginalization, nuclear fragmentation, and appearance of picnotic nuclei, especially when the highest concentrations of the product were used. Thus, permethrin induced early degeneration of this tissue characterized by significant changes in the structure of acinar cells and production of enzymes related to the cell death process, in addition to interfering directly in the genetic material of these cells.     

Morphological characterization of the nymphs Rhipicephalus sanguineus ticks (Latreille, 1806) (Acari: Ixodidae). Description of the testes, integument, Malpighian tubules, and midgut on the detachment day

This study presents the morpho-histological and histochemical characterization of the testes, integument, Malpighian tubules, and midgut of engorged Rhipicephalus sanguineus nymphs on the detachment day, showing the morphological and physiological characteristics to this phase in the life cycle of these individuals. The testis is constituted by germinative cells (only spermatogonia) with large, round-shaped and strongly stained nuclei which are organized into cysts by a thin layer of somatic cells. The integument consists of a cuticle subdivided into epicuticle (lipoprotein) and procuticle (glycoproteic), and a layer of epithelial cells which present glycolipoprotein elements. The procuticle presents two distinct regions: the exocuticle (next to the epicuticle) and the endocuticle (next to the epithelial layer). The Malpighian tubules present a simple epithelium with small flat and/or cubic cells, which form its wall and delimitates a lumen full of lipoprotein material. The midgut consists of an epithelial wall formed by two types of digestive cells, spent cells and empty digest cells, and by generative cells supported by a basal lamina and a thin layer of muscular tissue. This study described the main organs of engorged nymphs of R. sanguineus, to generate information that can help researchers to better understand the biology of these ectoparasites; which is fundamental for the development of compounds that are less aggressive to the environment. In addition, if the immature stages of the ticks are controlled, the number of adult ticks able to cause damages to the animals--and to the man as well--is also under control.     

一种真菌漆酶的克隆与序列分析

比较了几种不同真菌DNA提取方法对血红密孔菌基因组的提取的影响,根据GenBank中真菌漆酶氨基酸保守序列设计的简并引物,以血红密孔菌基因组总DNA为模板,扩增到一条长度为1084bp的DNA片段。通过BLAST比对,其基因序列与密孔菌属同源性最高,为99%,氨基酸同源性为88%,说明扩增的片段确为密孔菌漆酶基因。     

两株丝状真菌联合转化生产香草醛的研究

本文对黑曲霉(Aspergillus niger)CGMCC0774和朱红密孔菌(Pycnoporus cinnabarinus)CGMCC1115联合转化来源于米糠油脚的阿魏酸生产香草醛进行研究。通过60Co诱变黑曲霉菌株,得到耐较高底物浓度且不降解产物的突变株产香草酸2.48g/L;其转化液经过滤,调pH至5.0,补加5g/L葡萄糖,灭菌处理后,作为朱红密孔菌离菌体转化的底物溶液进行转化,得到1.07g/L香草醛。     

红曲霉Monascus sanguineus X1处理黄水的培养基优化及酯化液制备

黄水是白酒酿造的主要副产物之一,富含多种有机质,可用于制备酯化液促进白酒增香,提高其利用率。研究了一株高产酯化酶的红曲霉菌株Monascus sanguineus X1,以酯化酶活力为指标,从碳源、碳氮比、接种量和pH值4个方面对X1菌株的培养基进行单因素实验及正交试验优化;采用酯化酶液处理黄水,制备酯化液,研究了黄水、乙醇、己酸等酯化前体物质添加量对酯化液制备的影响。结果表明,该红曲霉优化发酵培养基组分(以100 mL计):大米粉7.0 g,大豆蛋白胨2.0 g,NaNO30.2 g,KH2PO40.15 g,MgSO4·7H2O 0.1 g,培养基初始pH值5.0,接种量10%(体积分数)。在此条件下,添加体积分数为10%的黄水和体积分数为4%的乙醇,酶活力可达745.80 U/mL。向酯化酶液中加入体积分数为0.8%的己酸和体积分数为20%的乙醇继续培养1 d后,酯化液中己酸乙酯的体积分数可达0.121%;而向酯化酶液中补加体积分数为10%的黄水后,己酸乙酯和乳酸乙酯的体积分数分别可达0.055%和0.032%。该结果旨在为将黄水资源用于白酒增香提供理论参考。     

Inhibitory action of neem aqueous extract (Azadirachta indica A. Juss) on the vitellogenesis of Rhipicephalus sanguineus (Latreille, 1806) (Acari: Ixodidae) ticks

The present study revealed unheard of data about the action of aqueous extracts of neem leaves (Azadirachta indica) on the vitellogenesis of Rhipicephalus sanguineus ticks, proving that these extracts in 10 and 20% concentrations do not have the potential to kill the females; however, in lower concentrations (10%) provokes great morphological alterations in germinative cells such as the emergence of extended cytoplasmic vacuolization areas as well as the fragmentation of the germinal vesicle, even in those oocytes which were in initial stages of development (I-III), showing that neem is a potent agent which acts impeding one of the main metabolic stages of the ticks, i.e., the reproduction. In oocytes in final stages of development (IV-V) azadirachtin (neem's active principle) caused significant reduction in the size and quantity of proteic granules of the yolk and the inversion of their localization where the smaller granules before inside the cell (normal oocyte) were posteriorly observed in the periphery, and the bigger ones in the central region. Thus, the study showed that the alterations found both in the oocytes and in the pedicel cells indicated that azadirachtin acts on the process of tick's reproduction and signalizes that this plant can be used in the future to control ticks with the advantage of not being aggressive to nontarget organism or the environment. Furthermore, data here obtained showed that the most significant efficiency of the aqueous extract of neem is related to the concentration of 10%, proving that higher doses would not be so efficient.     

Immobilization of Pycnoporus sanguineus laccase by metal affinity adsorption on magnetic chelator particles

BACKGROUND: Immobilized enzymes provide many advantages over free enzymes including repeated or continuous reuse, easy separation of the product from reaction media, easy recovery of the enzyme, and improvement in enzyme stability. In order to improve catalytic activity of laccase and increase its industrial application, there is great interest in developing novel technologies on laccase immobilization. RESULTS: Magnetic Cu²⁺‐chelated particles, prepared by cerium‐initiated graft polymerization of tentacle‐type polymer chains with iminodiacetic acid (IDA) as chelating ligand, were employed for Pycnoporus sanguineus laccase immobilization. The particles showed an obvious high adsorption capacity of laccase (94.1 mg g^?1 support) with an activity recovery of 68.0% after immobilization. The laccase exhibited improved stability in reaction conditions over a broad temperature range between 45 °C and 70 °C and an optimal pH value of 3.0 after being adsorbed on the magnetic metal‐chelated particles. The value of the Michaelis constant (K_m) of the immobilized laccase (1.597 mmol L^?1) was higher than that of the free one (0.761 mmol L^?1), whereas the maximum velocity (V_max) was lower for the adsorbed laccase. Storage stability and temperature endurance of the immobilized laccase were found to increase greatly, and the immobilized laccase retained 87.8% of its initial activity after 10 successive batch reactions. CONCLUSION: The immobilized laccase not only can be operated magnetically, but also exhibits remarkably improved catalytic capacity and stability properties for various parameters, such as pH, temperature, reuse, and storage time, which can provide economic advantages for large‐scale biotechnological applications of laccase. Copyright © 2007 Society of Chemical Industry     

Comparative study of Monascus sanguineus and its albino mutant based on molecular and chemical profiling

This study compared the wild strain of Monascus sanguineus to its albino mutant. The comparison was based on the variation at the molecular level, sporulation pattern and chemical profiling of fungal grown rice. Random amplification polymorphic DNA (RAPD) was used to perceive genetic variation and it was observed that among used primers, OPC-06 gave a major unique band of around 1.1 kb with the albino strain. In the scanning electron microscopy (SEM) images of the wild strain, large cleistothecium with ascospores was observed, whereas for albino, only branched hyphae and conidia were observed. Both strains had shown appreciable antioxidant activity and growth inhibition was observed against gram-positive and gram-negative bacterial strains. Chemical profiling of the fungal grown rice extract revealed that various sugars were produced by both the strains. The albino strain was found to be an efficient producer of fatty acids when compared to the wild strain. One unknown fatty acid was also discovered from albino fungal grown rice.     

Optimal parameters for cinnabarin synthesis by Pycnoporus sanguineus

Pycnoporus sanguineus was grown in potato dextrose broth, and the environmental parameters for cinnabarin synthesis were considered. The best growth conditions for improving the production of this substance by the fungus were: initial medium pH 9·0, incubation temperature 25°C, under light. ©1997 SCI