Uncommon life history and winter spawning of common carp (Cyprinus carpio) in a natural thermal spring, under temperate climate.

Common carp female generally matures at age 4-5 years old and spawns between April and July under the temperate climate. Contrary to a range of 0-28 °C of temperate freshwaters, the water temperature of Lake Hévíz (Hungary, Central Europe), the largest natural bathable thermal lake in the world, varies between 26 and 35 °C seasonally. The specific environmental conditions (continuously warm water and its individual chemical composition, special nutrient base, lack of natural lakeside spawning substrate compared to usual spawning grounds, continuous high human disturbance, etc.) suggest that the carp population here may also differ in reproductive characteristics from their counterparts in surrounding waters. Our findings suggest that the self-sustaining dwarf common carp population of Lake Hévíz matures 2 to 4 years earlier (at the age of one) and spawns 1 to 3 months before (between February and April, at 27-30 °C water temperature) than carp typically do in the temperate zone (16-20 °C). Successful winter spawning was verified by rearing larvae from the collected eggs and in situ induced propagation.

Acute effects of angler's groundbaits: nutrient flux to water column.

Although ground-baiting related nutrient loading has been widely studied, we do not know what proportion of these nutrients release into the water column, affecting primary production directly. We conducted short-term (24-h, 5-day) experiments at wide temperature range, in presence and absence of fish using fish meal-based (FM-GB) and plant-based groundbait (PB-GB), to assess the nitrogen (N) and phosphorus (P) fluxes from GB into the water column. Nitrogen release from unconsumed FM-GB was negligible in the first 3 days, then increased abruptly, releasing 32% of its total N content by the fifth day. In contrast, PB-GB acted as temporary sink for inorganic N forms. Considerable (18-21%) inorganic P release was observed in both GB types in the first twelve hours. Consumed GBs induced considerable inorganic N release and its rate increased with temperature. Particulate forms predominated the released N in PB-GB, suggesting impaired digestion. Phosphorus-dominated by particulate forms-release was similar or lower than in unconsumed GB. Based on our results, excessive use of GB-when high amount of it remains unconsumed-can enhance eutrophication in P-limited ecosystems. Although less digestible GBs may have less abrupt effect on the primary production, undigested nutrients remain unavailable for removal through fish harvest.

Efficacy of In Vitro Lithium Chloride Treatments on Dermacentor reticulatus.

Dermacentor reticulatus (Fabr., 1794) (Acari: Ixodidae) is parasite that spreads many diseases which are dangerous to humans and animals. Microelement lithium was found to have promising potential against the detrimental bee pest Varroa destructor. Furthermore, its effectiveness was confirmed against Dermanyssus gallinae, a major parasite of poultry, in vitro. In the present study, we investigated whether the efficacy of lithium chloride extends to other parasitic species, such as D. reticulatus. Our results revealed, for the first time, that the effectiveness of lithium chloride extends to D. reticulatus, confirmed to have 100% mortality at a relatively high minimum concentration of 1.38 M in vitro. The 24 h and 48 h median lethal concentration (LC50) values proved to be 0.654 M and 0.481 M, respectively, for this species. Our pilot study may contribute to a better understanding of the properties of lithium ion. Furthermore, it may elicit further studies aiming to reveal whether the different environmental mineral conditions may influence the D. reticulatus population. Further studies might reveal whether lithium has any possible veterinary relevance.

Lithium Chloride Shows Effectiveness against the Poultry Red Mite (Dermanyssus gallinae).

The poultry red mite (Dermanyssus gallinae) is the main pest of poultry, causing severe problems by being a vector of several animal and human pathogens. The number of miticides is few, and their efficacy in practice implies problems of residues and resistance; therefore, the demand for a new and safe agent is constant. The present publication investigated the effectiveness of lithium chloride under in vitro conditions on poultry red mites. This chemical currently appears to be one of the most promising alternatives to study amongst potential applicants to treat varroosis, a fatal disease of honey bees. In Experiment I, the previously used experimental doses (5.52 M, 2.76 M, 1.38 M) on Varroa mites confirmed their in vitro activity on the poultry red mite. Three event times (uncontrolled movement, immobilisation and death) were recorded to base the response to treatment for each concentration. In Experiment II, the LD 50 value was calculated, i.e., the value at which 50% of the mites were killed by the treatment. This Experiment showed that the LD50 of lithium chloride = 0.265 M in the poultry red mite. It is to note that the study remained restricted to in vitro confirmation of lithium chloride's effectiveness on the parasite. Thus, further extensive studies are needed to decide whether it has any relevance in practice against D. gallinae, and also to assess potential residue problems that could affect poultry products.

Comparison of Alternative Application Methods for Anti-Varroa Lithium Chloride Treatments.

Varroosis is one of the most dangerous threats to the bee industry but means of its treatment are still unsatisfactory. Lithium-based anti-Varroa treatments may provide an alternative, as this trace element can be a natural component of honey and is well tolerated by adult bees. However, it can be toxic to larvae and its use in beekeeping practice is not yet well understood. The present study aimed to investigate the efficacy of relevant application methods of acaricides used in beekeeping practice in brood-free conditions for lithium. Vaporisation proved to be an inefficient method of lithium treatment and killed only 9.9 ± 3.3% (mean ± SD) of mites in the hive. Lithium-impregnated paper strips showed moderate efficiency by killing 55.1 ± 26.2% of mites. The most effective way of applying lithium was the trickling method; different trickling treatments decreased the abundance of mites on average by 65 to 99.7%, depending on the applied dosage and the number of treatments. Repeated trickling treatments were more effective than single treatments, and they generally provided >90% efficiency. Experiments also proved that adding sugar to the trickling solution does not influence treatment efficiency. Thus, it is suggested that repeated and sugar-free trickling treatments with moderate lithium dosage could be the most rational methodology. Since lithium is not yet legalised in beekeeping practice, comprehensive studies are also needed to uncover the amount of lithium residue in bee products, depending on the treatment parameters.

Comparison of the Genetic Structure of Invasive Bigheaded Carp (Hypophthalmichthys spp.) Populations in Central-European Lacustrine and Riverine Habitats.

Bigheaded carps (bighead carp, Hypophthalmichthys molitrix, and silver carp, Hypophthalmichthys nobilis) and their hybrids play an important ecological and economic role in their original habitat, while their introduced stocks may pose serious ecological risks. To address questions about the persistence and invasiveness of these fish, we need to better understand their population structures. The genetic structures of bigheaded carp populations inhabiting Lake Balaton and the Tisza River were examined with ten microsatellite markers and a mitochondrial DNA marker (COI). The Lake Balaton stock showed higher genetic diversity compared with the Tisza River stock. Based on hierarchical clustering, the Tisza population was characterized only by only silver carps, while the Balaton stock included hybrid and silver carp individuals. All COI haplotypes originated from the Yangtze River. Based on the high genomic and mitochondrial diversity, along with the significant deviation from H-W equilibrium and the lack of evidence of bottleneck effect, it can be assumed that bigheaded carps do not reproduce in Lake Balaton. The present stock in Balaton may have originated from repeated introductions and escapes from the surrounding fishponds. The Tisza stock consists solely of silver carp individuals. This stock appears to have significant reproductive potential and may become invasive if environmental factors change due to climate change.

Changes in Lithium Levels in Bees and Their Products Following Anti-Varroa Treatment.

The biggest threat to beekeeping is varroosis caused by the mite Varroa destructor. Chemicals available to treat this fatal disease may present problems of resistance or inconsistent efficacy. Recently, lithium chloride has appeared as a potential alternative. To date, the amount of residue lithium treatments may leave in honeybee products is poorly understood. Honeybees were fed with 25 mM lithiated sugar syrup, which was used in earlier studies. The accumulation and elimination of the lithium were monitored in bees and their products for 22 days. Lithium concentration increased in the entire body of the bees to day 4 post-treatment and then recovered rapidly to the control level. Lithium exposure was found to affect uncapped honey in the short term (<16 days), but ripe (capped) honey measured at the end of the trial remained affected. On the other hand, lithium treatment left beeswax lithium-free. Based on these data, we propose that comprehensive research on harvested honey is needed to decide on the veterinary use of lithium.

Lithium chloride outperformed oxalic acid sublimation in a preliminary experiment for Varroa mite control in pre-wintering honey bee colonies.

Since lithium salts were demonstrated to be very effective for the potential control of Varroa destructor, a highly detrimental parasite of honey bee (Apis mellifera), no studies have been reported on their comparison with any commonly used varroicides in commercial bee colonies. In this study we compared the effectiveness of lithium chloride to that of oxalic acid, a widely used miticide. The results of the present study confirm that lithium has superior efficacy to oxalic acid sublimation both as a main or a supplementary pre-wintering treatment at moderate infestation levels, restricted to certain pre-wintering conditions. Considering its easy implementation in apicultural practice and its twofold mode of action, trickling would be the preferred way of administration after the use of lithium salts as varroicides is authorised.

Only one can remain? Environmental and spatial factors influencing habitat partitioning among invasive and native crayfishes in the Pannonian Ecoregion (Hungary).

Biological invasions have increasingly threatened indigenous species, influence metacommunity organisation and consequently, global biodiversity. World-wide expansion of non-indigenous crayfish (NICS) is associated with dramatic changes in species poor indigenous crayfish (ICS) assemblages challenging conservation planning. We analysed long-term changes of crayfish occurrences from the pre-invasion state, through the first appearance of NICS, to their intensive spread in Hungarian waters. Further, we analysed present-day crayfish metacommunity patterns for co-occurrences and influence of spatial and environmental factors. Historic data revealed a marked pre-invasion decline in indigenous noble crayfish Astacus astacus and stone crayfish Austropotamobius torrentium populations, but not in the narrow-clawed crayfish Pontastacus leptodactylus. Historic data provided no direct evidence for the impact of NICS on ICS, rather it supported that NICS often entered areas where ICS had been extinct or were not present at all. Crayfish species extremely rarely co-occurred which could indicate their strong competition and be related to utilization of empty sites by NICS. Crayfish metacommunities were predominantly spatially structured indicating the primary influence of ongoing invasion. Crayfish species also exhibited different environmental preferences mainly along the altitude and temperature gradients. We conclude that the invasion is still in the expanding phase and without an effective conservational program the future of ICS is doubtful in Hungary. Conservation policy should focus on the preservation and reintroduction of the stone and noble crayfishes in highland refugees. Expansion of NICS should be prevented in refugee areas by utilizing possibilities provided by natural and artificial barriers, and education and strict ban should be simultaneously applied to prevent further illegal releases by aquarists.

Contact Effect Contribution to the High Efficiency of Lithium Chloride Against the Mite Parasite of the Honey Bee.

Lithium chemicals have been proven to be very effective in eradicating Varroa destructor, the detrimental parasite of the honey bee; however, little is known about the side effects on brood and long term consequences on the colony. Earlier, it was proposed that the action mechanisms of lithium chloride do not include the contact mode. Here, we investigate this question using a paper strip test to demonstrate the concentration-dependent effectiveness of lithium in the contact mode of action, confirming that it is also a contact agent against the Varroa mite. According to our knowledge, this is the first report on the high varroicidal effect of lithium in the contact mode of action. Our findings may open up possibilities for novel ways of treatment (e.g., the use of lithiated strips) in the event that lithium salts become legal for use in apiculture.

Fusiform vateritic inclusions observed in European eel (Anguilla anguilla L.) sagittae.

Microscopic inclusions have been observed in 7 out of 106 European eel (Anguilla anguilla L.) sagittae using polarizing microscope and scanning electron microscope meanwhile the annual increments were studied to characterize the age structure of the population living in Lake Balaton. The presence of vaterite, a rare calcium carbonate polymorph was observed in these inclusions using Raman spectroscopy. Vateritic sagittae in wild fish are usually considered as symptom of physiological stress. The observed fusiform inclusions represent a new morphological type of vaterite inclusions in eel otolith. Two alternatives are hypothesized to explain their formation: 1) metabolic disorder, such as erroneous protein synthesis; 2) introduction of an alien protein into the eel's inner ear. The origin and physiological significance of this new morphological type of vateritic inclusions is still an open question. Same as whether it can be found in other species or specific only to eel otoliths.

Gut content microbiota of introduced bigheaded carps (Hypophthalmichthys spp.) inhabiting the largest shallow lake in Central Europe.

Studying the microbiota in the alimentary tract of bigheaded carps (Hypophthalmichthys spp.) gained special interest recently, as these types of investigations on non-native fish species may lead to a better understanding of their ecological role and feeding habits in an invaded habitat. For microbiological examinations, bigheaded carp gut contents and water column samples from Lake Balaton (Hungary) were collected from spring to autumn in 2013. Denaturing Gradient Gel Electrophoresis (DGGE) and pyrosequencing of the 16S rRNA gene were performed to reveal the composition. According to the DGGE patterns, bacterial communities of water samples separated clearly from that of the intestines. Moreover, the bacterial communities in the foreguts and hindguts were also strikingly dissimilar. Based on pyrosequencing, both foregut and hindgut samples were predominated by the fermentative genus Cetobacterium (Fusobacteria). The presence of some phytoplankton taxa and the high relative abundance of cellulose-degrading bacteria in the guts suggest that intestinal microbes may have an important role in digesting algae and making them utilizable for bigheaded carps that lack cellulase enzyme. In turn, the complete absence of typical heterotrophic freshwater bacteria in all studied sections of the intestines indicated that bacterioplankton probably has a negligible role in the nutrition of bigheaded carps.

Population Genetic Patterns of Threatened European Mudminnow (Umbra krameri Walbaum, 1792) in a Fragmented Landscape: Implications for Conservation Management.

The European mudminnow (Umbra krameri) is a Middle Danubian endemic fish species, which is characterised by isolated populations living mainly in artificial habitats in the centre of its range, in the Carpathian Basin. For their long term preservation, reliable information is needed about the structure of stocks and the level of isolation. The recent distribution pattern, and the population genetic structure within and among regions were investigated to designate the Evolutionary Significant, Conservation and Management Units (ESUs, CUs, MUs) and to explore the conservation biological value of the shrinking populations. In total, eight microsatellite loci were studied in 404 specimens originating from eight regions. The results revealed a pronounced population structure, where strictly limited gene flow was detected among regions, as well as various strengths of connections within regions. Following the results of hierarchical structure analyses, two ESUs were supposed in the Carpathian Basin, corresponding to the Danube and Tisza catchments. Our results recommend designating the borders of CUs in an 80-90km range and 16 clusters should be set up as MUs for the 33 investigated populations. How these genetic findings can be used to better allocate conservation resources for the long term maintenance of the metapopulation structure of this threathened endemic fish is discussed.

Genetic heterogeneity reveals on-going speciation and cryptic taxonomic diversity of stream-dwelling gudgeons (Teleostei, Cyprinidae) in the middle Danubian hydrosystem (Hungary).

Although stream-dwelling gudgeons (Cyprinidae, genus: Gobio) are widespread in Central Europe, the taxonomy of this group and the distribution of its species are still unexplored in detail. The aims of our study are to ascertain taxonomic composition and distribution of the former Gobio gobio superspecies in the inner area of the Carpathian Basin. Since the presence of cryptic species is suspected in this area, we examined the taxonomic and phylogenetic relationships of Central European Gobio taxa by sequencing the mitochondrial DNA control region (mtCR). Additionally, we characterized the genetic structure of 27 stream-dwelling gudgeon populations of this area by Amplified Fragment Length Polymorphism (AFLP). Results of mtCR analysis proved the presence of three species already known as G. obtusirostris (dominant in NW-Hungary), G. gobio (sporadic) and G. carpathicus (sporadic). Additionally, the analysis revealed the existence of one doubtful taxon, G. sp1 (dominant in NE-Hungary), and a new isolated haplogroup (dominant in SW-Hungary). Although Network analysis showed significant detachment among haplogroups, their genetic distances were quite small. Therefore Bayesian phylogenetic analysis showed weak nodal support for the branching pattern both for newly described haplotypes, and for the already accepted species. AFLP data showed distinct population structure and a clear pattern of isolation was revealed by distance of stocks. At the same time, level of separation was not affected by the altitudinal position of sites. Moreover we found three major clusters of populations which were separated according to hydrographic regions, and corresponded to the findings of mtCR analysis. Our results suggest the on-going speciation of gudgeons in the Carpathian Basin, however the separation of haplogroups seems to only be an intermediate phase. The discovered natural pattern seems to be only slightly influenced by anthropogenic impacts. Additionally our results put into question the suitability of the recently accepted within Gobio genus taxonomy.

Re-visiting phylogenetic and taxonomic relationships in the genus Saga (Insecta: Orthoptera).

Twelve of the 13 bushcricket species of the Saga genus are bisexuals and diploids, except the parthenogenetic and tetraploid bush cricket, Saga pedo. Despite a continuous research effort stretching through the 1900s, the taxonomic relationships of the Saga species are still disputed. In this study, our primary aim was to reveal natural relationships of the European Saga species and three of their Asian relatives, with special attention to the problematic taxonomy of two subspecies: S. campbelli campbelli and S. c. gracilis. Following a phylogenetic analysis of eight species, a comprehensive study was carried out on the above three taxa by using acoustic and morphometric approaches in parallel. Our phylogenetic data showed that European Saga species evolved from a monophyletic lineage. The geographical transitional species S. cappadocica was positioned between European and Asian lineages supporting the idea that the European Saga lineage originated phylogeographically from the Asian clade. The above results showed better agreement with the morphological data than with earlier ones based either on karyology or acoustic information only. After reviewing our data, we concluded that Saga pedo has most likely evolved from S. c. gracilis and not from S. rammei or S. ephippigera, as proposed by earlier studies. S. c. gracilis shares the same ITS2 haplotype with S. pedo, indicating that the latter could have evolved from populations of the former, probably through whole genome duplication. Based on acoustic and morphometric differences, we propose to elevate the two subspecies, S. campbelli campbelli and S. c. gracilis, to species level status, as Saga gracilis Kis 1962, and Saga campbelli Uvarov 1921. The present work sets the stage for future genetic and experimental investigations of Saginae and highlights the need for additional comprehensive analysis involving more Asian Saga species.

Age- and size-specific patterns of heavy metals in the organs of freshwater fish Abramis brama L. populating a low-contaminated site.

Concentrations of cadmium, copper, mercury, lead and zinc were determined by atomic absorption spectrophotometry in the muscle, gill and liver of bream Abramis brama L. to study the relationship between the heavy metal load of fish and their age and size, and the seasonal variation of pollutant loads. Fish were collected from the Western basin of Lake Balaton (Hungary) in October 1999 and May 2000. The average metal concentrations of different organs varied in the following ranges: Cd 0.42-2.10; Cu 1.77-56.2; Hg 0.01-0.19; Pb 0.44-3.24; Zn 10.9-82.5 microg g(-1) dry weight. The highest Cd, Cu, Pb and Zn concentrations were detected in the gill or liver of fish, whereas the highest Hg concentrations were measured in the muscle. In the liver of bream for cadmium, copper and mercury the Pearson correlation analysis revealed positive associations related to age and size (length, net weight), as well as for the mercury load of all three investigated organs. In the muscle and gill the copper, lead and zinc concentrations, similarly to the lead and zinc concentrations of the liver, the associations related to age and size were negative. The correlations between the heavy metal concentrations of organs and the individual condition factors of fish samples proved to have opposite trends compared to those related to the age and size of fish. The seasonal variations in the heavy metal load of bream could be attributed rather to the seasonal change in the condition factor of fish than to variations in the pollutant load of the site.