Boguś M I, Kedra E, Bania J, Szczepanik M, Czygier M, Jabłoński P, Pasztaleniec A, Samborski J, Mazgajska J, Polanowski A
W. Stefański Institute of Parasitology, Polish Academy of Sciences, 00-818 Warszawa, Poland.
J Insect Physiol. 2007 Sep;53(9):909-22. doi: 10.1016/j.jinsphys.2007.02.016. Epub 2007 Mar 12.
The resistance of Galleria mellonella, Dendrolimus pini, and Calliphora vicina larvae against infection by the enthomopathogen Conidiobolus coronatus was shown to vary among the studied species. Exposure of both G. mellonella and D. pini larvae to the fungus resulted in rapid insect death, while all the C. vicina larvae remained unharmed. Microscopic studies revealed diverse responses of the three species to the fungal pathogen: (1) the body cavities of D. pini larvae were completely overgrown by fungal hyphae, with no signs of hemocyte response, (2) infected G. mellonella larvae formed melanotic capsules surrounding the fungal pathogen, and (3) the conidia of C. coronatus did not germinate on the cuticle of C. vicina larvae. The in vitro study on the degradation of the insect cuticle by proteases secreted by C. coronatus revealed that the G. mellonella cuticle degraded at the highest rate. The antiproteolytic capacities of insect hemolymph against fungal proteases correlated well with the insects' susceptibility to fungal infection. The antiproteolytic capacities of insect hemolymph against fungal proteases correlated well with the insects' susceptibility to fungal infection. Of all the tested species, only plasmatocytes exhibited phagocytic potential. Exposure to the fungal pathogen resulted in elevated phagocytic activity, found to be the highest in the infected G. mellonella. The incubation of insect hemolymph with fungal conidia and hyphae revealed diverse reactions of hemocytes of the studied insect species. The encapsulation potential of D. pini hemocytes was low. Hemocytes of G. mellonella showed a high ability to attach and encapsulate fungal structures. Incubation of C. vicina hemolymph with C. coronatus did not result in any hemocytic response. Phenoloxidase (PO) activity was found to be highest in D. pini hemolymph, moderate in G. mellonella, and lowest in the hemolymph of C. vicina. Fungal infection resulted in a significant decrease of PO activity in G. mellonela larvae, while that in the larvae of D. pini remained unchanged. PO activity in C. vicina exposed to fungus slightly increased. The lysozyme-like activity increased in the plasma of all three insect species after contact with the fungal pathogen. Anti E. coli activity was detected neither in control nor in infected D. pini larvae. No detectable anti E. coli activity was found in the control larvae of G. mellonella; however, its exposure to C. coronatus resulted in an increase in the activity to detectable level. In the case of C. vicina exposure to the fungus, the anti E. coli activity was significantly higher than in control larvae. The defense mechanisms of D. pini (species of economic importance in Europe) are presented for the first time.
研究表明,大蜡螟、松毛虫和红头丽蝇幼虫对昆虫病原冠状耳霉感染的抗性在不同研究物种间存在差异。大蜡螟和松毛虫幼虫接触该真菌后会迅速死亡,而所有红头丽蝇幼虫均未受伤害。显微镜研究揭示了这三个物种对真菌病原体的不同反应:(1)松毛虫幼虫的体腔被真菌菌丝完全覆盖,无血细胞反应迹象;(2)受感染的大蜡螟幼虫在真菌病原体周围形成黑化包囊;(3)冠状耳霉的分生孢子在红头丽蝇幼虫的角质层上不萌发。对冠状耳霉分泌的蛋白酶降解昆虫角质层的体外研究表明,大蜡螟角质层降解速度最快。昆虫血淋巴对真菌蛋白酶的抗蛋白水解能力与昆虫对真菌感染的易感性密切相关。在所有测试物种中,只有浆细胞表现出吞噬潜力。接触真菌病原体导致吞噬活性升高,在受感染的大蜡螟中发现其吞噬活性最高。昆虫血淋巴与真菌分生孢子和菌丝的孵育揭示了所研究昆虫物种血细胞的不同反应。松毛虫血细胞的包囊潜力较低。大蜡螟血细胞表现出高附着和包囊真菌结构的能力。红头丽蝇血淋巴与冠状耳霉孵育未导致任何血细胞反应。发现酚氧化酶(PO)活性在松毛虫血淋巴中最高,在大蜡螟中中等,在红头丽蝇血淋巴中最低。真菌感染导致大蜡螟幼虫PO活性显著降低,而松毛虫幼虫的PO活性保持不变。接触真菌后,红头丽蝇的PO活性略有增加。与真菌病原体接触后,所有三种昆虫血浆中的溶菌酶样活性均增加。在对照和受感染的松毛虫幼虫中均未检测到抗大肠杆菌活性。在大蜡螟对照幼虫中未发现可检测到的抗大肠杆菌活性;然而,其接触冠状耳霉后活性增加到可检测水平。在红头丽蝇接触真菌的情况下,其抗大肠杆菌活性显著高于对照幼虫。首次介绍了松毛虫(在欧洲具有经济重要性的物种)的防御机制。