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Antifungal activity of various essential oils against Diplodia seriata and Botryosphaeria dothidea isolates from apple fruit
No full textThe Botryosphaeriaceae family encompasses a range of morphologically diverse fungi that can live as pathogens, endophytes or saprobes. Fungi from this family can also be found in healthy tissue parts of the plant and usually cause diseases after the plant is exposed to stressful conditions (post-harvest or heavy rain). Species from this family are considered aggressive pathogens. The use of naturally derived bioactive compounds is one of the most promising ecological alternatives and has advantages over synthetic fungicides, particularly for postharvest pathogen control. Essential oils (EOs) are volatile compounds extracted from various plants and are known for antifungal, antiseptic and antimicrobial effects with minimal toxic residue on the harvested fruits and crops.
Three Diplodia seriata and three Botryosphaeria dothidea isolates were obtained from symptomatic apple fruits. Isolates were identified based on morphological characteristics and by sequence analysis of ITS region and two genes (partial translation elongation factor 1-alpha gene (TEF1) and partial β-tubulin gene (TUB)). Antifungal activity of four commercially available EOs: thyme (Thymus vulgaris), rosemary (Rosmarinus officinalis), lavender (Lavandula spp.) and lemongrass (Cymbopogon citratus) was tested using fumigant macrodilution method. The experiment was conducted in glass Petri dishes (diameter: 90 mm) on PDA medium with mycelial fragments (diameter: 6 mm) placed at the center of each dish. Five different volumes of selected EOs were pipetted onto filter paper cuttings attached to the inner side of the dish covers to achieve final air phase concentrations of 0.02, 0.05, 0.07, 0.09 and 0.12 µl ml⁻¹ of air. The dishes were inverted, sealed with parafilm to prevent external gas exchange and incubated at 25°C for 5 days. Distilled water was used as negative control. The percentage growth inhibition (PGI) was determined by measuring the radial growth of the isolates exposed to different oil concentrations and comparing them with control dishes. The experiment was replicated three times.
Thyme essential oil demonstrated the highest antifungal activity against both fungi, achieving a 100% inhibition rate at oil concentration of 0.05 µl ml⁻¹ of air phase. Lemongrass essential oil also showed strong inhibition potential, with PGI ranging from 85.1% at 0.02 µl ml⁻¹ of air phase to 100% at 0.07 µl ml⁻¹ of air phase against D. seriata. For B. dothidea, PGI values ranged from 73.6% at 0.02 µl ml⁻¹ of air phase to 100% at 0.09 µl ml⁻¹ of air phase. Moderate antifungal activity was demonstrated by rosemary EO with PGI values ranging from 34.2% against D. seriata and from 20.6% against B. dothidea at the lowest concentration, up to 53.8% against D. seriata and to 59.7% against B. dothidea at the highest concentration tested. Lavender EO showed the lowest inhibition potential with PGI range from 0.0% to 13.8% for D. seriata and from 5.4% to 13.2% for B. dothidea.
These findings indicate that thyme and lemongrass EO exhibit a supreme antifungal activity against D. seriata and B. dothidea isolates, achieving a 100% mycelial growth inhibition at relatively low EO concentrations, what makes those oils the most promising candidates for further in vivo testing
Antagonistički mikroorganizmi za biološko suzbijanje Sclerotinia spp
No full textVrste roda Sclerotinia prouzrokuju belu trulež biljaka koja predstavlja
veliki problem u biljnoj proizvodnji širom sveta. Kao patogeni koji se održa-
vaju u zemljištu dugi niz godina, vrlo su problematični za suzbijanje. Kako bi
se smanjio rizik od nastanka ekonomski značajnih šteta, većina proizvođača
koristi integralni sistem zaštite koji podrazumeva kombinovanu primenu ag-
rotehničkih, hemijskih i bioloških mera suzbijanja. Agrotehničke mere kao što
su plodored, smanjenje gustine biljaka i mehaničko suzbijanje korova imaju
važnu ulogu u zaštiti useva od prouzrokovača bele truleži. Zbog nedostatka
otpornih sorti gajenih biljaka i širokog spektra biljaka domaćina, upotreba
fungicida u suzbijanju Sclerotinia spp. predstavlja dominantni način suzbija-
nja. Međutim, primena fungicida predstavlja opasnost kako za životnu sredinu
i poljoprivredne proizvođače, tako i za potrošače koji bivaju izloženi ostacima
pesticida u biljnim proizvodima. Poslednjih godina primena bioloških mera u
suzbijanju vrsta iz ovog roda dobija sve veći značaj. U radu su sistematizova-
na aktuelna saznanja o prouzrokovačima bele truleži, njihovom ciklusu razvo-
ja i simptomima koje izazivaju na različitim domaćinima. Detaljno je opisana
primena antagonističkih mikroorganizama (gljive, bakterije, aktinomicete,
mikovirusi) za biološko suzbijanje i dat pregled biofungicida registrovanih u
svetu i našoj zemlji za zaštitu useva od vrsta roda Sclerotinia
Pesticidi i fitomedicina = Pesticides and Phytomedicine
No full textGlavni i odgovorni urednik naučnog časopisa Pesticidi i fitomedicina = Pesticides and Phytomedicin
Potential of different strains of Steinernema feltiae (Filipjev) to control Lycoriella ingenua (Dufour) and impact on mushroom yield
No full textThe study was focused on improvement of integrated management strategy against the mushroom
fly Lycoriella ingenua (Dufour) on the white button mushroom Agaricus bisporus (Lange)
Imbach. Impact of different strains of entomopathogenic nematodes (EPN) Steinernema feltiae
(Filipjev), commercial and two indigenous from Serbia (K2, P9), on regulation of mushroom
sciarid fly and the influence on mushroom yield was evaluated. Experiment was conducted in air-
conditioned mushroom growing chamber. Commercially produced phase III mushroom substrate
were placed into 24 plastic containers (w 31 × d 21.5 × h 13 cm). Each plastic container was
inoculated with 20 last instar larvae of L. ingenua. EPN were produced on last larval instars of
Galleria mellonella L. and less than a week old infective juveniles (IJ) were used in the
experiment. Nematode suspension of each of the three EPN strains containing 50000 IJ were
added on the substrate surface in each plastic container on the day before casing, at the casing
time and seven days after casing. Containers were held in the insect rearing cages and density of
adults of L. ingenua was observed weekly using yellow sticky traps inside each cage. Preliminary
results of ongoing experiment showed the highest efficacy in pest control of commercial EPN
strain, followed by two native strains. No statistically significant differences were found in
efficacy between two Serbian strains. Also, no statistically significant differences in mushroom
yield were observed among control and all treatments using different strains of EPNs
Altering Microbial Communities in Substrate to Stimulate the Growth of Healthy Button Mushrooms
No full textGreen mould, caused by Trichoderma aggressivum, is one of the major fungal diseases of
button mushrooms. The main problems in chemical disease control include a lack of effective agents,
the occurrence of pathogen resistance to pesticides, and the harmful impact on the environment.
In an attempt to find a solution, the interaction between two beneficial microorganisms, Bacillus
amyloliquefaciens B-241 (an antifungal agent) and Streptomyces flavovirens A06 (a yield stimulant),
was investigated in vivo. The synergy factor (SF) was calculated as a ratio between the observed
and expected impact on the yield or efficacy of disease suppression after artificial inoculation with
T. aggressivum. The highest control of T. aggressivum was achieved by joint application of the two beneficial
microorganisms. The additive interaction between microorganisms in efficacy against the
pathogen was revealed. The largest yield was obtained in mushroom beds sprayed with the two beneficial
microorganisms combined (B-241 80% and A06 20%). Regarding the impact on the yield,
synergistic interaction between the two microorganisms was confirmed (SFs were 1.62 or 1.52). The
introduction of optimized microbial combinations could create new possibilities for biorational edible
mushroom protection, with improved yield and quality and reduced risks to human health and
the environment
The predators attacking soft scales (Hemiptera: Coccomorpha: Coccidae) in Serbia
No full textThis study aimed to determine the diversity of species of predators attacking scale insects (Hemiptera: Coccidae) in
Serbia. Over a four-year period, at 25 locations and on 25 host plants infested with 16 species of Coccidae, a total of 18,050
predators were collected and reared. Among them, were 16 species belonging to 7 families and 5 insect orders (Coleoptera,
Dermaptera, Diptera, Hemiptera, and Neuroptera). Six species, Harmonia axyridis, Hippodamia variegata, Propylea quatuordecimpunctata,
Forfi cula auricularia, Deraeocoris ruber and Oxycarenus lavaterae, were recorded for the fi rst time feeding on coccid
pests in Serbia and O. lavaterae was fi rst recorded being zoophagous. The most common species was Exochomus quadripustulatus,
which was recorded feeding on all 16 species of soft scale insects, while the dominant species was Anthribus nebulosus, with
a total of 17,385 individuals collected and reared on fi ve species of soft scale. Considering the potential of the predators as biocontrol
agents for suppressing scale insects, the data obtained indicate the need to understand predator diversity, which is one of
the prerequisites for using these predators for controlling coccid pests
Harnessing Bacillus spp. for targeted biocontrol of Sclerotinia sclerotiorum
No full textSclerotinia sclerotiorum, a devastating fungal
pathogen, poses significant challenges in
managing diseases across various plant species,
including economically significant crops
worldwide. This plant-pathogenic fungus, distributed
across temperate, tropical, and arid
regions, causes severe crop damage, leading to
substantial yield losses. Given the limitations of
synthetic agents for its suppressing, attention
has turned towards biological control methods,
particularly the selection of beneficial microorganisms.
Therefore, the aim of this research was
to detect antagonistic effect of bacterial isolates
against S. sclerotiorum. Seventeen bacterial isolates
isolated from soil (PAM2.1 – PAM 2.17) were
screened for their antagonistic activity against
this plant pathogen in vitro, on PDA medium.
Additionally, bacterial traits significant for the
biocontrol, such as the production of siderophores,
amylase and cellulases were assessed
on appropriate solid media, while the presence
of genes coding for antibiotics (fengycin, surfactin,
iturin C, subtilin, and bacilysin) was assessed
by PCR method. Further, for the most effective
isolate the amplification of DNA sequence coding
for 16S rRNA gene was done using P0/ P6
primer pair. For the identification, the obtained
sequences were compared to those available in
the National Center for Biotechnology Information
(NCBI) database. Out of 17 tested bacterial
isolates, only one (isolate PAM 2.2) showed
antagonistic effect against S. sclerotiorum, with
inhibition of mycelial growth of about 71%. The
same isolate showed the ability to produce siderophores,
amylase and cellulase (CMCase). Further,
the presence of genes coding for surfactin,
subtilin and bacillomycin was detected for the isolate
PAM2.2. Based on the NCBI BLASTn analysis
of the 16S rRNA gene sequence, isolate PAM2.2
was identified as Bacillus spp. This research highlights
isolate PAM2.2 as a promising candidate
for biological control against S. sclerotiorum,
demonstrating significant antagonistic effects,
production of relevant enzymes and antibiotics,
thus offering a potential sustainable solution for
disease management in agricultur
Molecular characterization of Leptospira species in wild rats in Belgrade, Serbia
No full textЦиљ – истраживања је био испитивање преваленције и молекуларних
карактеристика Leptospira 5рр. у Београду. Посебан фокус био је на примени
молекуларних техника у типизацији појединих врста ове бактерије на нивоу
сероваријетста. Током 2020., 2021. и 2022. године, укупно је сакупљено 344 (186
женки и 158 мужјака) лешева сивих пацова (Ratfus norvegicus) у околини града
Beorpana, претежно у урбаним и приградским стаништима. Применом
различитих молекуларних дијагностичких алата анализирани су узорци бубрега
пацова. Применом Real-time PCR специфично усмереног на детекцију гена
lipL32 (за разликовање патогених и сапрофитских Leptospira зрр.) утврђено је
присуство ове бактерије у 108 узорака, а затим су позитивни узорци
искоришћени за умножавање серије гена: adk, icdd, lipL32, lipL41, rrs2 и secY
чијом анализом је вршена типизација. Истраживањем је откривено присуство
две засебне Leptospira spp. (L. interrogans и L. kirschneri) и три сероваријетета
(L. interrogans serovar Copenhageni, L. interrogans serovar Icterohaemorrhagiac и
L. kirschneri serovar Mozdok). Ово истраживање довело је до првог открића
присуства L. kirschneri serovar Mozdok у Србији и доприноси бољем
разумевању молекуларне разноврсности и преваленцији врста из рода
Leptospira. Налази истичу кључну улогу молекуларних техника у разјашњавању
епидемиолошких карактеристика и карактеризацији Leptospira spp. и доприносе
развијању сфикасних мера контроле ове зоонотске болести. Истраживање
наглашава важност континуираног надзора и молекуларне карактеризације
Leptospira 5рр. у популацијама урбаних пацова као средства 3a боље разумевање
динамике преноса лептоспирозе доприносећи развоју циљаних стратегија
заштите јавног здравља и примена превентивних мера. .This research aimed 10 delve into the prevalence and molecular characteristics of
Leptospira species in Belgrade, Serbia. The focus was particularly on employing
advanced molecular and multilocus sequence typing (MLST) techniques to bridge the
knowledge gap concerning the variety and spread ог Leptospira species across this
region. During 2020,2021 and 2022, а total of 344 (186 female and 158 male) carcasses
ог Norway rats (Rattus norvegicus) угете collected in the broad environs of Belgrade
City, pmdominam\y in their urban and suburban habitats. Through а detailed molecular
examination of rats’ Kidney samples, the study leveraged different molecular diagnostic
tools. These included real-time PCR, specifically targeting the lipL32 gene (for the
differentiation between pathogenic and saprophytic Leptospira species), alongside
sequence—based typing frameworks that utilized a series of genes: adk, icdA, lipL32,
lipL41, rrs2, and secY. The investigation unveiled е presence of two distinct
Leptospira species (L. inferrogans и L. kirschneri) and three unique serovars (L.
interrogans serovar Copenhageni, L. interrogans serovar Icterohaemorrhagiae и L.
Fkirschneri serovar Mozdok) within the Belgrade vicinity, marking the first-time
detection of L. kirschneri serovar Mozdok in Serbia. This interesting finding enriches
the existing body of knowledge regarding the molecular diversity and prevalence of
Leptospira species in the агеа, contributing significantly to the scientific community's
understanding of such pathogens. These findings highlight the critical role of advanced
molecular typing techniques in elucidating ће epidemiological patterns and
characterizations ог Leptospira species and offer cssential nsights that are instrumental
in devising effective control measures and interventions against this zoonotic disease.
The research underscores the importance of continuous surveillance and molecular
characterization of Leptospira in urban rat populations as а means to better understand
the dynamics of leptospirosis transmission, ultimately contributing to the development
of targeted public health strategies and preventative measures
Water emulsion of the essential oil of Nepeta rtanjensis Diklić et Milojević: potential use as a bioherbicide
No full textPlant protection with natural products is a new trend in environmentally friendly agriculture. Nepeta rtanjensis
Diklić et Milojević is an endemic and critically endangered plant species in Serbia. We explored the phytotoxic potential of
a water emulsion of Nepeta rtanjensis essential oil (NrEO) with high amounts of trans,cis-nepetalactone on five weeds. The
most sensitive was Stellaria media (L.) Vill., as NrEO (from 0.013% to 0.1%) completely inhibited germination. Germination
of Amaranthus retroflexus L. and Artemisia vulgaris L. was completely inhibited at the highest applied concentrations
of NrEO (0.1%), while Ambrosia artemisiifolia L. germination was reduced to 48% at the same concentration of NrEO. The
most tolerant species was Cephalaria transsylvanica (L.) Schrad. ex Roem. & Schult. as the final germination rate at the
highest applied concentration of NrEO (0.1%) was 81%, like the control (82%). To our knowledge, this is the first time the
interaction of essential oils on the germination and growth of A. vulgaris and C. transsylvanica is reported. The germination
dynamics of S. media in pots with soil were significantly inhibited when the soil was initially treated with the highest
applied concentration of the NrEO water emulsion (1%). Spraying S. media seedlings with NrEO significantly inhibited
growth parameters (shoot height, shoot fresh weight, and the number of nodes) at the higher applied concentrations of
NrEO (0.5% and 1%). Therefore, the water emulsion of the essential oil of N. rtanjensis could be potentially developed for
use in the control of invasive and allergenic weeds
First record of plum seed wasp Eurytoma schreinerei Schreiner (Hymenoptera,Eurytomidae) in Serbia
No full textMany eurytomid wasps (Eurytomidae) are plant parasites, with four Paleartic species of
particular economic importance to stone fruit cultivation: Eurytoma amygdali Enderlein, E.
mas/ovskii Nikolskaya, E. samsonowi Vasiljev and f. schreineri Schreiner. To date, these species
have not been recorded in Serbia. However, in 2023, a new species of eurytomid wasp attacking
plum was detected for the first time in the Pomoravlje District in Central Serbia.
Field surveys were conducted in 2023-2024 across the major plum-producing regions to
evaluate the distribution of the newly detected wasp species, characterize its infestation
symptoms, and assess the extent of damage to different plum varieties in Serbia. A total of 45
samples of fallen fruits from various plum varieties were collected across 13 administrative
districts. In the laboratory, the infested fruits were incubated until the emergence of adults.
Monitoring of the seasonal dynamics of the wasp was carried out at two sites in the Pomoravlje
district using yellow sticky traps. The samples were preserved in 70% ethanol for morfological
analyses and 96% ethanol for DNA extraction. The wasps were identified using morphological
keys based on the characters of adult females and larvae. Total genomic DNA was extracted from
individual specimens using the E.Z.N.A. Tissue DNA Kit (USA), following the manufacturer's
protocol. Morphological identification was further supported by sequence analysis of the mitochondrial cytochrome c oxidase subunit 1 (COi) gene barcode. The obtained sequences
were compared with those available in the NCBI database.
Based on morphological characteristics and COi analyses, the wasp was identified as E.
schreineri. This species was found in 16 plum samples, from the three regions: Norteast Serbia
(Nort Banat District), Central Serbia (Pomoravlje District) and Southeast Serbia (Pirot District). The
wasp attacks the fruits of various plum cultivars, with the most significant damage observed in
the Caca nska rodna and 5tan/eyvarieties, where infestation rates ranged from 10-96%.
This univoltine species overwinters as larva within the stones of fallen fruits beneath plum
trees. In the climatic condition of Serbia, adults emerge from the fallen fruits in early spring
( beginning of April). After mating, females lay eggs in newly formed plum fruit. Infested fruits
fall prematurely to the ground, during June and July, particularly during the the BBCH 76-80
phenophase . No visible damage is observed in the epicarp and mesocarp as the larvae feed on
the endosperm.
This is the first report of this non-native pest species in Serbia. Future studies will assess its
economic impact, investigate its biology, and map its distribution to develop effective
management strategies for plum production