Identification of gut bacteria in worker and soldier castes of the termite Anacanthotermes ahngerianus

Anacanthotermes ahngerianus termitining ishchi va soldat kastalaridan kultivatsiya qilinadigan ichak bakteriyalari ajratib olindi va MALDI-TOF MS hamda Gram usulida bo‘yash yordamida tahlil qilindi. O‘n beshta sof izolyat olindi: yettitasi ishchilardan, sakkiztasi soldatlardan. Ishchilardan olingan izolyatlar Enterobacter (57,1%) va Bacillus (28,6%) avlodlariga mansub deb topildi; bitta izolyat (14,3%) identifikatsiya qilinmay qoldi. Soldatlardan olingan barcha izolyatlar Acinetobacter avlodiga mansub ekanligi aniqlandi. Natijalar tajriba sharoitida ikki kasta o‘rtasida kultivatsiya qilinadigan ichak bakteriyalarining taksonomik tarkibida farqlar borligini ko‘rsatadi.

FUNDAMENTAL VA AMALIY MIKROBIOLOGIYANING BUGUNGI HOLATI VA RIVOJLANISH ISTIQBOLLARI: YOSH OLIMLAR NIGOHI Respublika ilmiy-amaliy anjumani · Toshkent, 2026-yil 30-oktabr

I FUNDAMENTAL VA AMALIY MIKROBIOLOGIYA DOI 10.66073/ReFocus-Conf/3010029

IDENTIFICATION OF GUT BACTERIA IN WORKER AND SOLDIER CASTES OF THE TERMITE ANACANTHOTERMES AHNGERIANUS Utaganov Muslimbek Ulug‘bek o‘g‘li1, Zakiryayeva Saidaxon Ikramovna2,3, Xolmatov B.R.1 1 Institute of Zoology, Academy of Sciences of the Republic of Uzbekistan, Tashkent, Uzbekistan 2 Institute of Microbiology, Academy of Sciences of the Republic of Uzbekistan, Tashkent, Uzbekistan 3 Republican Scientific and Production Center for Termite Control under the Institute of Zoology, Academy of Sciences of the Republic of Uzbekistan, Tashkent, Uzbekistan E-mail: muslimotaganov78@gmail.com Abstract: Cultivable gut bacteria from worker and soldier castes of Anacanthotermes ahngerianus were isolated and analysed using MALDI-TOF MS and Gram staining. Fifteen pure isolates were obtained: seven from workers and eight from soldiers. Worker isolates were assigned to Enterobacter (57.1%) and Bacillus (28.6%); one isolate (14.3%) remained unidentified. All soldier isolates were assigned to Acinetobacter. The results indicate differences in the taxonomic composition of cultivable gut bacteria between the two castes under the experimental conditions. Keywords: Anacanthotermes ahngerianus; gut bacteria; worker caste; soldier caste; MALDI-TOF MS; Gram staining. Introduction Gut microorganisms play an important role in the digestion of cellulose-rich food by termites [1]. Studying the gut microbiota of Anacanthotermes ahngerianus is therefore relevant to understanding its feeding biology, identifying cellulose-degrading microorganisms and developing the biological basis of termite control. Termite guts harbour complex microbial communities involved in lignocellulose degradation. Because worker and soldier castes differ in colony functions and feeding characteristics, their cultivable gut bacterial communities may also differ. This study aimed to identify bacterial isolates from the guts of both castes using MALDI-TOF MS and to compare their taxonomic composition. Materials and methods Termite specimens were collected from Beruniy District, Republic of Karakalpakstan, in collaboration with the Republican Scientific and Production Center for Termite Control. Ten termites from each caste were used. Guts were dissected under aseptic conditions and homogenised in sterile phosphate buffer. The homogenates were diluted in sterile 0.9% NaCl solution and plated on meat-peptone agar. Cultures were incubated at 30 °C for 72 h. Morphologically distinct colonies were repeatedly subcultured to obtain pure cultures. Isolates were characterised using conventional methods, including Gram staining, and identified by matrix-assisted laser desorption/ionisation time-of-flight mass spectrometry (MALDI-TOF MS) [2]. Results Fifteen pure bacterial isolates were obtained: seven from workers (WG-1, WG-2, WG-4, WG-6, WG-5/1, WG-5/2 and WG-5/3) and eight from soldiers (SG-1 to SG-8). Worker isolates WG-5/2 and WG-5/3 were Gram-positive; the remaining worker isolates and all soldier isolates

FOR CITATION Utaganov M.U., Zakiryayeva S.I., Xolmatov B.R. Identification of gut bacteria in worker and soldier castes of the termite Anacanthotermes ahngerianus // Proceedings of the Republican Scientific and Practical Conference “Status and Development Prospects of Fundamental and Applied Microbiology: The Viewpoint of Young Scientists”. – Tashkent, 2026. – P. 110–111. DOI: https://doi.org/10.66073/ReFocus-Conf/3010029

FUNDAMENTAL VA AMALIY MIKROBIOLOGIYANING BUGUNGI HOLATI VA RIVOJLANISH ISTIQBOLLARI: YOSH OLIMLAR NIGOHI 30.10.2026

were Gram-negative. These reactions were consistent with the general phenotypic characteristics of the bacterial taxa assigned by MALDI-TOF MS (Table 1). Table 1. MALDI-TOF MS results for gut bacterial isolates from worker and soldier castes of Anacanthotermes ahngerianus Identification Caste Isolate code MALDI-TOF MS identification score Worker WG-5/1 Unidentified 1.47 Worker WG-5/2 Bacillus megaterium 2.42 Worker WG-5/3 Bacillus subtilis 2.04 Worker WG-4 Enterobacter cloacae 2.05 Worker WG-6 Enterobacter cloacae 2.26 Worker WG-1 Enterobacter asburiae 1.95 Worker WG-2 Enterobacter cloacae 2.21 Soldier SG-1 Acinetobacter junii 1.98 Soldier SG-2 Acinetobacter parvus 1.88 Soldier SG-3 Acinetobacter junii 1.94 Soldier SG-4 Acinetobacter junii 1.99 Soldier SG-5 Acinetobacter parvus 2.10 Soldier SG-6 Acinetobacter pittii 1.86 Soldier SG-7 Acinetobacter junii 2.31 Soldier SG-8 Acinetobacter parvus 2.04 Discussion Among worker isolates, MALDI-TOF MS assignments comprised Enterobacter (57.1%) and Bacillus (28.6%), while one isolate (14.3%) remained unidentified. All eight soldier isolates were assigned to Acinetobacter. These findings indicate differences in the taxonomic composition of cultivable bacteria between termite castes under the experimental conditions. Conclusion A total of 15 pure bacterial isolates were recovered and analysed by MALDI-TOF MS. Worker isolates included representatives of Bacillus and Enterobacter, whereas all soldier isolates were assigned to Acinetobacter. Assessing their cellulolytic activity and confirming species-level identifications using molecular genetic methods are necessary to establish their functional roles in termite nutrition.