Antimicrobial Peptides (AMP) in the Cell-Free Culture Media of Xenorhabdus budapestensis and X. szentirmaii Exert Anti-Protist Activity against Eukaryotic Vertebrate Pathogens including Histomonas meleagridis and Leishmania donovani Species

András Fodor; Claudia Hess; Petra Ganas; Zsófia Boros; János Kiss; László Makrai; Károly Dublecz; László Pál; László Fodor; Anna Sebestyén; Michael G Klein; Eustachio Tarasco; Manjusha M Kulkarni; Bradford S McGwire; Tibor Vellai; Michael Hess

doi:10.3390/antibiotics12091462

Antimicrobial Peptides (AMP) in the Cell-Free Culture Media of Xenorhabdus budapestensis and X. szentirmaii Exert Anti-Protist Activity against Eukaryotic Vertebrate Pathogens including Histomonas meleagridis and Leishmania donovani Species

Antibiotics (Basel). 2023 Sep 19;12(9):1462. doi: 10.3390/antibiotics12091462.

Authors

András Fodor¹, Claudia Hess², Petra Ganas², Zsófia Boros^{1

3}, János Kiss³, László Makrai⁴, Károly Dublecz⁵, László Pál⁵, László Fodor⁶, Anna Sebestyén⁷, Michael G Klein⁸, Eustachio Tarasco⁹, Manjusha M Kulkarni¹⁰, Bradford S McGwire¹⁰, Tibor Vellai¹, Michael Hess²

Affiliations

¹ Department of Genetics, Institute of Biology, Eötvös Loránd University, Pázmány Péter. sétány 1C, H-1117 Budapest, Hungary.
² Clinic for Poultry and Fish Medicine, Department for Farm Animals and Veterinary Public Health, University of Veterinary Medicine (Vetmeduni Vienna), 1210 Vienna, Austria.
³ Agribiotechnology and Precision Breeding for Food Security National Laboratory, Department of Microbiology and Applied Biotechnology, Institute of Genetics and Biotechnology, Hungarian University of Agriculture and Life Sciences, Páter Károly utca 1, H-2100 Gödöllő, Hungary.
⁴ Autovakcina Kft., H-1171 Budapest, Hungary.
⁵ Institute of Physiology and Nutrition, Georgikon Campus, Hungarian University of Agriculture and Life Sciences (MATE), Deák Ferenc utca 16, H-8360 Keszthely, Hungary.
⁶ Department of Microbiology and Infectious Diseases, University of Veterinary Medicine, H-1143 Budapest, Hungary.
⁷ First Department of Pathology and Experimental Cancer Research, Semmelweis University, H-1085 Budapest, Hungary.
⁸ USDA-ARS & Department of Entomology, The Ohio State University, 13416 Claremont Ave, Cleveland, OH 44130, USA.
⁹ Department of Soil, Plant and Food Sciences, University of Bari "Aldo Moro", Via Amendola 165/A, 70126 Bari, Italy.
¹⁰ Division of Infectious Diseases, Department of Internal Medicine, The Ohio State University, Columbus, OH 43210, USA.

Abstract

Anti-microbial peptides provide a powerful toolkit for combating multidrug resistance. Combating eukaryotic pathogens is complicated because the intracellular drug targets in the eukaryotic pathogen are frequently homologs of cellular structures of vital importance in the host organism. The entomopathogenic bacteria (EPB), symbionts of entomopathogenic-nematode species, release a series of non-ribosomal templated anti-microbial peptides. Some may be potential drug candidates. The ability of an entomopathogenic-nematode/entomopathogenic bacterium symbiotic complex to survive in a given polyxenic milieu is a coevolutionary product. This explains that those gene complexes that are responsible for the biosynthesis of different non-ribosomal templated anti-microbial protective peptides (including those that are potently capable of inactivating the protist mammalian pathogen Leishmania donovanii and the gallinaceous bird pathogen Histomonas meleagridis) are co-regulated. Our approach is based on comparative anti-microbial bioassays of the culture media of the wild-type and regulatory mutant strains. We concluded that Xenorhabdus budapestensis and X. szentirmaii are excellent sources of non-ribosomal templated anti-microbial peptides that are efficient antagonists of the mentioned pathogens. Data on selective cytotoxicity of different cell-free culture media encourage us to forecast that the recently discovered "easy-PACId" research strategy is suitable for constructing entomopathogenic-bacterium (EPB) strains producing and releasing single, harmless, non-ribosomal templated anti-microbial peptides with considerable drug, (probiotic)-candidate potential.

Keywords: CFCM (cell-free conditioned culture media); Histomonas meleagridis; Leishmania; Photorhabdus (P. luminescens TT01); X. budapestensis; X. innexii; X. szentirmaii; Xenorhabdus; entomopathogenic nematode-symbiont bacteria (EPB); extracellular avian pathogens; kinetoplastid protozoa parasite; non-ribosomal anti-microbial peptides (NR-AMP); probiotic potential.

Grants and funding

These project has partly been funded by Prof. Vellai, OTKA (Hungaria Research Fund) Grant Number: K132439 at the Department of Genetics, Eövös University Another source of funding: Dr. János Kiss, OTKA (Hungarian Research Fund) Grant Number: K 128203. The experiments in Gödöllő (J.K., Zs.B.) were also supported by RRF-2.3.1-21-2022-00007 from NKFIH (Nemzeti Kutatási, Fejlesztési és Innovációs Hivatal, https://nkfih.go, Hungary). The “EMA-EMC International Project” started at the Department of Genetics, Eötvös University; several other labs have been involved but have also been terminating here, although a significant part has been accomplished somewhere else. The Histomonas experiment was part of the Hungarian–Austrian international cooperation and supported by the TÁMOP-4.2.2/B-10/1-2010-0025 project based on the financial support of the State of Hungary and the EU. The CFCM preparations for Histomonas experiments were supported by the Hess–Dublecz project (recently a joint unit of the Hungarian University of Agriculture and Life Sciences (MATE). The Leishmania experiments were supported by the bench money of Bradford McGwire in Columbus, OH, USA.