Impact of tourism on the vigilance behavior of the Cyprus rock agama (Laudakia cypriaca).

Escape behavior is a common antipredator strategy among wild animals. Here, we investigated the effect of four factors on the vigilance behavior of the endemic Cyprus rock agama (Laudakia cypriaca). Flight initiation distance (FID, the minimum distance to which an observer can approach a lizard before it flees) was measured in relation to the type of location (tourist vs. nontourist area), the observer's starting distance, air temperature, and substrate temperature. We collected data for 39 agamas in tourist areas and 34 of these lizards in nontourist areas. As a whole, the mean starting distance was 10.5 m and the FID was 3.6 m. The average substrate temperature was 34.0°C and the average air temperature 29.6°C. Only the type of area affected the agamas' escape decisions with FID being 1.8 m shorter in tourist areas than in nontourist areas (2.7 m vs. 4.5 m). This is probably due to the habituation of lizards to the presence of humans in the former areas. This study shows that tourism strongly affects the behavior of lizards, which may have consequences for the functioning of the population. Tourists can increase the safety of lizards by creating a human shield to deter predators. Once the tourist season is over, lizards may become more vulnerable to predators.

Delayed egg-laying in Red-backed Shrike Lanius collurio in relation to increased rainfall in east-central Poland.

Climate change has affected the breeding parameters of many animal species. In birds, most studies have focused on the effects of temperature on clutch phenology and clutch size. The long-term influence of other weather factors, including rainfall, on breeding parameters have been analysed much less often. Based on a 23-year dataset and 308 broods, we documented shifts in the timing of breeding, clutch size and mean egg volume in a long-distance migrant, the Red-backed Shrike Lanius collurio, from a central European population. We found a 5-day shift towards delayed breeding, but no differences in brood size or egg volume during those 23 years. The GLM analysis showed that the mean May temperature had a positive influence on the clutch initiation date, whereas the number of days with rain delayed laying. During the period 1999-2021, there was no change in the mean May temperature, but total precipitation and the number of days with rain in May increased. Thus, delayed nesting in this population was probably due to the increase in rainfall during this period. Our results provide a rare example of delayed nesting in birds in recent years. Predicted changes in the climate make it difficult to assess the long-term impact of global warming on the viability of Red-backed Shrike populations in east-central Poland.

Flavonoids and Phenolic Acids Content in Cultivation and Wild Collection of European Cranberry Bush Viburnum opulus L.

Guelder rose (Viburnum opulus L.) is known for its health benefits. V. opulus contains phenolic compounds (flavonoids and phenolic acids), a group of plant metabolites with wide biological activities. They are good sources of natural antioxidants in human diets owing to their prevention of the oxidative damage responsible for many diseases. In recent years, observations have shown that an increase in temperature can change the quality of plant tissues. So far, little research has addressed the problem of the common impact of temperature and place of occurrence. Towards a better understanding of phenolics concentration that could indicate their potentials as therapeutic agents and towards predicting and controlling the quality of medicinal plants, the aim of this study was to compare phenolic acids and flavonoids content in the leaves of cultivation and wild collection V. opulus, and to examine the impacts of temperature and place of occurrence on their content and composition. Total phenolics were determined using the spectrophotometric method. Phenolic composition of V. opulus was determined using high-performance liquid chromatography (HPLC). The following hydroxybenzoic acids there were identified: gallic, p-hydroxybenzoic, syringic, salicylic, benzoic, as well as hydroxycinnamic acids: chlorogenic, caffeic, p-coumaric, ferulic, o-coumaric and t-cinnamic. The analysis of extracts from V. opulus leaves has indicated the presence of the following flavonoids: flavanols: (+)-catechin and (-)-epicatechin; flavonols: quercetin, rutin, kaempferol, myricetin; and flavones: luteolin, apigenin and chrysin. The dominant phenolic acids were p-coumaric and gallic acids. The major flavonoids found in V. opulus leaves were myricetin and kaempferol. Temperature and plant location affected the concentration of tested phenolic compounds. The present study shows the potential of naturally grown and wild V. opulus for the human.

Differences in Oxidative Stress Markers and Antioxidant Enzyme Activities in Black Bean Aphid Morphs (Aphis fabae Scop.) Fed on the Primary Host Viburnum opulus L.

Changes in the level of oxidative stress markers-superoxide anion radical (O2-), hydrogen peroxide (H2O2) and malondialdehyde (MDA) and the activity of antioxidant enzymes-superoxide dismutase (SOD), catalase (CAT) and ascorbate peroxidase (APX) in the black bean aphid occurring on the primary host (viburnum plants) were studied. Among the aphid morphs, the lowest contents of O2-, H2O2 and MDA were noted for winged adults (alatae), which were also characterized by the highest activity of antioxidant enzymes. These metabolic features indicate the adaptation of winged morphs to the colonization of new host plants. During spring migration, an increase in the content of oxidative stress markers and antioxidant enzyme activities in wingless females (fundatrigeniae) was observed. The significance of the biochemical adaptation of the black bean aphid to its winter host is discussed.

Expression of Thioredoxin/Thioredoxin Reductase System Genes in Aphid-Challenged Maize Seedlings.

Thioredoxins (Trxs) and thioredoxin reductases (TrxRs) encompass a highly complex network involved in sustaining thiol-based redox homeostasis in plant tissues. The purpose of the study was to gain a new insight into transcriptional reprogramming of the several genes involved in functioning of Trx/TrxR system in maize (Zea mays L.) seedlings, exposed to the bird cherry-oat aphid (Rhopalosiphum padi L.) or the rose-grass aphid (Metopolophium dirhodum Walk.) infestation. The biotests were performed on two maize genotypes (susceptible Zlota Karlowa and relatively resistant Waza). The application of real-time qRT-PCR technique allowed to identify a molecular mechanism triggered in more resistant maize plants, linked to upregulation of thioredoxins-encoding genes (Trx-f, Trx-h, Trx-m, Trx-x) and thioredoxin reductase genes (Ftr1, Trxr2). Significant enhancement of TrxR activity in aphid-infested Waza seedlings was also demonstrated. Furthermore, we used an electrical penetration graph (EPG) recordings of M. dirhodum stylet activities in seedlings of the two studied maize varieties. Duration of phloem phase (E1 and E2 models) of rose-grass aphids was about three times longer while feeding in Waza plants, compared to Zlota Karlowa cv. The role of activation of Trx/TrxR system in maintaining redox balance and counteracting oxidative-induced damages of macromolecules in aphid-stressed maize plants is discussed.

Changes in the content of thiol compounds and the activity of glutathione s-transferase in maize seedlings in response to a rose-grass aphid infestation.

The rose-grass aphid (Methopolophium dirhodum Walk.) is a major pest of maize (Zea mays L.), but little is known about the biochemical interactions between M. dirhodum and its host plant. Thiol compounds and glutathione S-transferase (GST) play a crucial role in the defense responses of maize to biotic stress factors, including aphids. The purpose of this research was to evaluate the impact of M. dirhodum herbivory on the total thiol (TT), protein bound thiol (PT), reduced glutathione (GSH) and oxidized glutathione (GSSG) contents as well as the activity of GST in three varieties of Z. mays (Zlota Karlowa, Ambrozja and Plomyk), that were classified as aphid-susceptible, aphid-relatively resistant and aphid-resistant, respectively. The earliest and strongest aphid-triggered alterations in the levels of TT, PT and GSH, and the greatest induction of GST activity, were recorded in the resistant Plomyk seedlings in relation to the relatively resistant Ambrozja and the susceptible Zlota Karlowa.

Aphid-Triggered Changes in Oxidative Damage Markers of Nucleic Acids, Proteins, and Lipids in Maize (Zea mays L.) Seedlings.

Prior experiments illustrated reactive oxygen species (ROS) overproduction in maize plants infested with bird-cherry-oat (Rhopalosiphum padi L.) aphids. However, there is no available data unveiling the impact of aphids feeding on oxidative damages of crucial macromolecules in maize tissues. Therefore, the purpose of the current study was to evaluate the scale of oxidative damages of genomic DNA, total RNA and mRNA, proteins, and lipids in seedling leaves of two maize genotypes (Zlota Karlowa and Waza cvs-susceptible and relatively resistant to the aphids, respectively). The content of oxidized guanosine residues (8-hydroxy-2'-deoxyguanosine; 8-OHdG) in genomic DNA, 8-hydroxyguanosine (8-OHG) in RNA molecules, protein carbonyl groups, total thiols (T-SH), protein-bound thiols (PB-SH), non-protein thiols (NP-SH), malondialdehyde (MDA) and electrolyte leakage (EL) levels in maze plants were determined. In addition, the electrical penetration graphs (EPG) technique was used to monitor and the aphid stylet positioning and feeding modes in the hosts. Maize seedlings were infested with 0 (control), 30 or 60 R. padi adult apterae per plant. Substantial increases in the levels of RNA, protein and lipid oxidation markers in response to aphid herbivory, but no significant oxidative damages of genomic DNA, were found. Alterations in the studied parameters were dependent on maize genotype, insect abundance and infestation time.

Mechanism of entomotoxicity of the Concanavalin A in Rhopalosiphum padi (Hemiptera: Aphididae).

The toxicity effect of Concanavalin A (Canavalia ensiformis lectin, ConA) to bird cherry-oat aphid, Rhopalosiphum padi L. (Hemiptera: Aphididae), was investigated in the laboratory by using artificial diets containing ConA concentrations. Bird cherry-oat aphid performance was affected by the presence of Con A in artificial diets. The lectin added into the liquid diet increased the prereproductive period, mortality, and the average time of generation development (T) and decreased fecundity and the intrinsic rate of natural increase (rm). In attempt to unravel the mode of action of ConA, the interaction of the lectin with insect gut and the effect of ConA on feeding behavior were investigated. Extract of gut of treated grain aphid demonstrated DNA fragmentation, and this was accompanied with an increase in caspase 3 activity. Moreover, addition of ConA to the sucrose-agarose gels reduced salivation and passive ingestion of fluids from the gel. The results indicate that the insecticidal activity of ConA on R. padi may involve effects on death of the gut epithelial cells and effects on feeding behavior. This can be employed to create plants that are resistant to aphids.

Expression profiling of selected glutathione transferase genes in Zea mays (L.) seedlings infested with cereal aphids.

The purpose of this report was to evaluate the expression patterns of selected glutathione transferase genes (gst1, gst18, gst23 and gst24) in the tissues of two maize (Zea mays L.) varieties (relatively resistant Ambrozja and susceptible Tasty Sweet) that were colonized with oligophagous bird cherry-oat aphid (Rhopalosiphum padi L.) or monophagous grain aphid (Sitobion avenae L.). Simultaneously, insect-triggered generation of superoxide anion radicals (O2•-) in infested Z. mays plants was monitored. Quantified parameters were measured at 1, 2, 4, 8, 24, 48 and 72 h post-initial aphid infestation (hpi) in relation to the non-infested control seedlings. Significant increases in gst transcript amounts were recorded in aphid-stressed plants in comparison to the control seedlings. Maximal enhancement in the expression of the gst genes in aphid-attacked maize plants was found at 8 hpi (gst23) or 24 hpi (gst1, gst18 and gst24) compared to the control. Investigated Z. mays cultivars formed excessive superoxide anion radicals in response to insect treatments, and the highest overproduction of O2•- was noted 4 or 8 h after infestation, depending on the aphid treatment and maize genotype. Importantly, the Ambrozja variety could be characterized as having more profound increments in the levels of gst transcript abundance and O2•- generation in comparison with the Tasty Sweet genotype.

Are naringenin and quercetin useful chemicals in pest-management strategies?

The effects of two polyphenolic flavonoids (flavanone naringenin and flavonol quercetin) on development, fecundity, and mortality of the pea aphid, Acyrthosiphon pisum Harris (Hemiptera: Aphididae), were determined in vitro, on an artificial diets. Also determined in vitro (DC EPG method), on sucrose-agarose gels, were the effects of flavonoids on the probing and feeding behavior of adult apterae. When added to a liquid diet, higher concentrations of studied flavonoids increased the developmental time, the pre-reproductive period, and mortality and decreased fecundity and the intrinsic rate of natural increase of A. pisum. In most events associated with stylet activity (as indicated by EPG waveform g-C), differences in probing behavior did not statistically differ between the control gel and those with flavonoids; quercetin at 10, 100, and 1,000 µg cm-3 prolonged the number of gel penetrations; and quercetin only at 10,000 µg cm-3 prolonged the time the first g-C waveform was observed. Addition of flavonoids to the gels generally reduced passive ingestion from fluids of the gels (EPG waveform g-E2). At higher concentrations (>1,000 µg cm-3) the flavonoids completely stopped salivation (EPG waveform g-E1) and passive ingestion from fluids of the gels (EPG waveform g-E2). In events associated with active ingestion (EPG waveform g-G), however, differences in feeding behavior did not statistically differ between the control gel and those with flavonoids. The present findings demonstrate detrimental effects of the flavanone naringenin and flavonol on the behavior of the pea aphid. This can be employed in a biotechnological projects for plant breeding resistant to herbivores, including aphids.

Antifeedant activity of luteolin and genistein against the pea aphid, Acyrthosiphon pisum.

Electrical penetration graphs (DC EPG) were used to monitor the feeding behavior of the pea aphid, Acyrthosiphon pisum Harris (Hemiptera: Aphididae) exposed to the flavonoids luteolin and genistein in artificial diets. The EPG patterns generated by aphids feeding on plants were used to interpret the patterns generated on the artificial diets. Addition of flavonoids to the diets generally prolonged the period of stylet probing (as indicated by EPG pattern d-C), reduced salivation (as indicated by pattern d-E1) and passive ingestion (as indicated by pattern d-E2), and also delayed the onset of salivation and passive ingestion. At higher concentrations (≥100 µg cm(-3) for luteolin, ≥1,000 µg cm(-3) for genistein), the flavonoids completely stopped salivation and passive ingestion. In most events associated with active ingestion (EPG pattern d-G), however, differences in feeding behavior did not statistically differ between the control diet and those with flavonoids; luteolin, and genistein only at 10 µg cm(-3) prolonged the time until the first d-G pattern was observed. The current findings demonstrate detrimental effects of the isoflavone genistein and the flavone luteolin on the feeding behavior of the pea aphid, A. pisum. This can be employed to create plants which are resistant to aphids and other herbivores.

Antimetabolic effect of phytohemagglutinin to the grain aphid Sitobion avenae fabricius.

The insecticidal activity of plant lectins against a wide range of insect species have been intensively studied. Understanding the mechanism of the toxicity of lectins is one of the studied aspects. In the present research, the first step was determine the effect of phytohemagglutinin (PHA) on the development, fecundity and mortality of grain aphid. Next, the effect of PHA lectin on the activity of such enzymes as: α- and ß-glucosidases, alkaline (AkP) and acid (AcP) phosphatases, aminopeptidase N and cathepsin L involved in the metabolism of sugar, phosphorus and proteins of an adult apterae aphids was investigated. The PHA lectin added into the liquid diet increased the pre-reproductive period, mortality of Sitobion avenae, the time of generation development and decreased its fecundity and the intrinsic rate of natural increase. In addition, activity of α-glucosidase, alkaline phosphatase and aminopeptidase N of adult apterae exposed to PHA were reduced. The results indicate that the insecticidal activity of PHA on S. avenae may involve changes in activity of the enzymes in the midgut and it may be part of its toxicity.

Molecular evidence of Anaplasma phagocytophilum and Babesia microti co-infections in Ixodes ricinus ticks in central-eastern region of Poland.

The aim of the study was to elucidate the distribution of Anaplasma phagocytophilum and Babesia microti co-infection in Ixodes ricinus populations within the central-eastern region of Poland. The prevalence of analysed tick-borne human pathogens in single and polymicrobial infections in I. ricinus ticks were analysed using the conventional and nested PCR techniques. A total number of 1,123 questing tick individuals (291 females, 267 males and 565 nymphs) were collected at different ecosystems (municipal parks, suburban forests, and woodlands). In the presented study, 95 samples of ticks (8.5%) were infected with A.phagocytophilum, 3.1% (n=35) with B. microti, whereas the co-existence status of these human pathogens was detected in 1.8% (n=20) of all tested samples. It has been demonstrated that the prevalence of co-infection status was the highest among females of I. ricinus (11 samples, 3.8%), whereas the lowest within tested nymphs (5 samples, 0.9%). Ticks collected at city parks in Warsaw and suburban areas of this town characterized the highest prevalence of co-infections (3.3 and 4.8%, respectively). Furthermore, it was established that co-infection rates of ticks inhabiting woodlands within Kampinos National Park and Nadbuzanski Landscape Park were similar and reached the levels of 1.4% (n=5) and 1.1% (n=4), respectively.

Induced proteolysis within the bird cherry leaves evoked by Rhopalosiphum padi L. (Hemiptera, Aphidoidea).

The objectives of this study were to elucidate the impact of bird cherry-oat aphid (Rhopalosiphum padi L.) feeding on functioning of the proteolytic machinery in bird cherry leaves. Biochemical analyses proved that R. padi feeding in tissues of primary host stimulated activity of the two major fractions of proteinases (extracted at the optimal pH values: 5.0 and 7.0). Additionally, it has been demonstrated that aphids' feeding on bird cherry led to a decline in levels of albumins and globulins (main protein fractions in P. padus leaves). The opposite tendency, regarding the amounts of these protein fractions was ascertained at the phase of disappearance of R. padi population on tested shoots. Furthermore, it is reported that an increase in activity of the analysed enzymes and a decline in the content of tested protein fractions, were proportional to density of aphid individuals developing on P. padus side shoots. It is hypothesized that long-term R. padi feeding may lead to intensifying the catabolic processing of proteins by the activated proteolytic machinery in bird cherry leaves. The multi-level biological functions of endogenous plant proteinases and their significance in triggering the defense reactions in aphid-infested plant tissues are discussed.

Biochemical markers of oxidative stress within tissues of cereal aphids.

Effects of some dietary o-dihydroxyphenols on biochemical markers of oxidative stress within tissues of Sitobion avenae (Fabricius), (Homoptera, Aphididae) and Rhopalosiphum padi (Linnaeus), (Homoptera, Aphididae) has been studied. Among the studied aphid morphs the highest concentration of total thiols, hydrogen peroxide and lipid peroxidation products (TBARS) was noted for winged adults (alatae). Higher content of H2O2 was observed within tissues of the oligophagous species R. padi while the monophagous species S. avenae had higher TBARS and total thiols concentration. Aphids exposed the dietary pro-oxidative o-dihydroxyphenols (quercetin, caffeic acid, chlorogenic acid) demonstrated significantly lower concentration of total sulfhydryls and higher level of TBARS and H2O2 than the control ones. Among the studied compounds, chlorogenic acid showed the strongest effect on the level of thiols and TBARS while caffeic acid caused the highest accumulation of the hydrogen peroxide within the aphid tissues.

Deterrence and toxicity of plant saponins for the pea aphid acyrthosiphon pisum harris.

Plant saponins are widely distributed among plants and have a wide range of biological properties. Three alfalfa saponins - zanhic acid tridesmoside, 3GlcA, 28AraRhaXyl medicagenic acid glycoside, and 3GlcA, 28AraRha medicagenic acid glycoside - were tested for their settling inhibition effects on feeding behavior of the aphid Acyrthosiphon pisum using the electrical penetration graph method. Application of saponins to artificial diets affected the insects' probing behavior. In general, saponins incorporated into sucrose-agarose gels significantly reduced the number of aphid probes and extended their duration. Lower saponin concentrations (50 ppm) extended aphid activity and corresponded to phloem sap ingestion. In contrast, higher concentrations (100 ppm) strongly reduced aphid ability to ingest phloem and xylem sap.