Sains Malaysiana 34(1): 7-16 (2005)

 

Teknik Pengesanan Spora Nosema bombycis dan Kesan

Jangkitan pada Larva Plutella xylostella

(Detection Techniques for Spores of Nosema bombycis and Effects

of Infection to the Larva of Plutella xylostella)

 

 

Zainal-Abidin B.A.H., Rosnizar M. Jarnil, Noran Majid, Idris A.B.

Pusat Pengajian Biosains & Bioteknologi

Pusat Pengajian Sains Sekitaran & Sumber Alam

Fakulti Sains & Teknologi

Universiti Kebangsaan Malaysia

43600 UKM Bangi, Selangor D.E. Malaysia

 

Sajap A.S.

Fakulti Perhutanan Universiti Putra Malaysia

43400 UPM Serdang, Selangor D.E.

 

Hussan A.K.

Malaysian Agricultural Research and Development Institute (MARDI)

P.O. BOX 12301, G.P.O

50774 Kuala Lumpur Malaysia

 

 

ABSTRAK

 

Beberapa teknik pewarnaan biasa telah digunakan untuk mengesan peringkat hidup Nosema bombycis (khususnya spora) yang terdapat pada larva Plutella xylostella (rama-rama belakang intan, DBM) yang terjangkit begitujuga kesan jangkitan spora ini ke atas larva telah dikaji. Larva instar 1,2,3 dan 4 DBM yang bebas daripadajangkitan spora telah dijangkiti dengan empat kepekatan spora (407150, 41420, 4260 dan 420 spora/μl) secara berasingan dengan membiarkan larva ini memakan makanan buatan yang telah dicemari dengan kepekatan spora tersebut. Pemprosesan tisu dengan teknik pewarnaan, penentuan kepekatan spora daripada larva dan cerapan langsung telah dilakukan dalam tempoh 24, 48 dan 72 j selepas jangkitan. Didapati teknik pewarnaan Gram, Giemsa, hematoksilin dan trikrom lebih sesuai untuk mengesan spora, sporon dan meron daripada tisu larva yang terjangkit berbanding teknik 'good Pasteur'. Kesan utama jangkitan spora ialah kematian larva yang bergantung kepada kepekatan spora yang digunakan dan didapati ins tar yang lebih muda adalah lebih rentan terhadap jangkitan ini berbanding instar yang lebih matang. Kepekatan spora 407150, 41420 dan 4260 spora) μl boleh menyebabkan kematian manakala kepekatan spora terendah (420 spora/)1.l) tidak menyebabkan kematian. Kadar kematian instar yang lebih muda lebih tinggi (p<0.001 ) berbanding instar yang lebih matang. Nilai LC50 dan LC95 bagi instar 1 dan 2 lebih rendah berbanding instar 3 dan 4, selepas 48 dan 72 j jangkitan. lni menunjukkan bahawa kepekatan spora yang lebih tinggi diperlukan untuk membunuh instar yang bersaiz lebih besar dan lebih matang. Bilangan spora terendah diperolehi pada tempohjangkitan 24 j manakala bilangan spora tertinggi diperolehi pada tempoh jangkitan 72 j. Kajian histologi ke atas tisu larva terjangkit mendapati kerosakan terjadi pada tisu lemak dan usus yang menyebabkan kematian. Hasil kajian ini menunjukkan bahawa peringkat hidup N bombycis khususnya spora dapat dikesan dengan baik menerusi teknik pewarnaan yang biasa dan kebolehjangkitan parasit ini dan kesannya kepada larva DBM telah diketahui.

 

Kata kunci: Nosema bombycis, Plutella xylostella

 

 

ABSTRACT

 

Several conventional staining techniques were employed to detect the life stages (in particular the spores) of Nosema bombycis from infected larvae of Plutella xylostella (diamondback moth, DBM) and the effects of the infection of N. bombycis on the larvae were studied. Larval instars 1, 2, 3 and 4 of the DBM were infected with four different spore concentrations (407150, 41420, 4260 and 420 spores/μl) accordingly by allowing them to feed on artificial diet previously inoculated with the respective spore concentrations. Larval tissues were processed for staining and the number of spores were counted and direct observations on the larvae were carried out at 24, 48 and 72 h post infection. The Gram s, Giemsa's, haematoxylin and trichrome staining techniques were more superior in detecting the spores, sporonts and meronts than the good Pasteurs. The main effect of the infection was mortality which was dose-dependent and that the younger ins tars were more susceptible to infection than the older ones. Spore concentrations of 407150, 41420 and 4260 spores/μl) caused death to the instars whereas the dose of 420 spores)μl was unable to kill the larval instars. The mortality rate of the younger instars was significantly higher (p<0.001) than the older ones. The LC50 and LC95 values of instar 1 and 2 were lower than those of the instar 3 and 4 after 48 and 72 h post infection. This showed that high spore concentrations were needed to kill the bigger size and matured instars. Histological studies on the infected larvae indicated that the infection caused severe cellular damages in fat tissues and the intestine leading to death. Results of this studies showed that life stages of N. bombycis in particular the spores were detected effectively using conventional staining techniques and the infectivity and the effects of the infection on the larval tissues of DBM were also established.

 

Keywords: Nosema bombycis, Plutella xylostella

 

 

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