Vol.:(0123456789) 1 3
Journal of Thermal Analysis and Calorimetry
https://doi.org/10.1007/s10973-020-10011-7
Estimation of microbiological contamination of maize seeds using
isothermal calorimetry
Andrzej Skoczowski
1
· Sebastian W. Przemieniecki
2
· Jakub Oliwa
3
· Monika Kula‑Maximenko
4
· Magdalena Rys
4
·
Iwona Stawoska
1
· Stanisław Karpiński
5
Received: 3 July 2019 / Accepted: 29 June 2020
© The Author(s) 2020
Abstract
The storage of maize seed intended for industrial purposes in foil silo bags is associated with microbiological contamina-
tion of the material by bacteria and fungi. This results in the loss of a part of the raw material or its deterioration and causes
fnancial losses. In this paper, the relationship between the number of microorganisms colonizing maize seeds as well as
changes in heat fow has been proved. For this purpose, the heat fow and total metabolic heat emission values (total heat)
were analyzed. Calorimetric measurements of seed samples were made using the TAM III isothermal calorimeter. The seed
samples with the same heat fow curves shape were homogenized, and microbial DNA was isolated from them. The quantita-
tive real-time polymerase chain reaction (qPCR) was performed for detecting the main group of microorganisms colonizing
maize seeds. It has been shown that in the case of seeds less afected by Bacillus and fungi (including yeast), the heat fow
(in the range of 0–360 min) has a falling shape, while for more infected it grows almost linearly. The more infected maize
seeds also show signifcantly higher values of total heat emission. The described research can be used to quickly assess the
degree of seed contamination.
Keywords Bacillus spp. · Bacteria · Fungi · Heat fow · Zea mays
Introduction
Long-term storage of maize seeds (intended for example
for the production of bioethanol) in plastic silo bags creates
problems of technological and economic nature. Microbial
contamination of the material by bacteria and fungi present
on the seed coat negatively afects the fermentation pro-
cesses and deterioration of the quality of the fnal product
(e.g., presence of mycotoxins, ammonia, butyric acid). In
addition, the reduction in the technological quality of seeds,
and consequently the loss of a part of the raw material,
causes signifcant fnancial losses. For this reason, moni-
toring the quality of the biological material taken is very
important [1, 2].
Microbiological analysis of environmental samples based
on microbial cultures (for example on Petri dishes) is time-
consuming and their results are often ambiguous. A better
alternative is molecular methods such as: DNA markers,
metagenomics analysis, DNA barcoding, molecular phyloge-
netic or using PCR. However, the disadvantage of methods
based on DNA analysis is the inability to recognize live cells
from dead ones as well as the variable efciency of DNA
extraction. An additional aspect is the need to design primers
and probes and optimize the hybridization conditions [3, 4].
For this reason, we have decided to use isothermal calorim-
etry to monitor the degree of microbial contamination of
seeds, without the need of long-term and expensive analysis.
* Andrzej Skoczowski
andrzej.skoczowski@up.krakow.pl
1
Institute of Biology, Pedagogical University of Cracow,
Podchorążych 2, 30-084 Kraków, Poland
2
Department of Entomology Phytopathology and Molecular
Diagnostics, University of Warmia and Mazury in Olsztyn,
Prawocheńskiego 17, 10-720 Olsztyn, Poland
3
Department of Chemistry and Biochemistry, Institute
of Basic Sciences, University of Physical Education, Jana
Pawła II 78, 31-571 Kraków, Poland
4
The Franciszek Górski Institute of Plant Physiology Polish
Academy of Sciences, Niezapominajek 21, 30-239 Kraków,
Poland
5
Department of Plant Genetics, Breeding and Biotechnology,
Warsaw University of Live Sciences, Nowoursynowska 159,
02-776 Warsaw, Poland