First serological and molecular evidence on the endemicity of Anaplasma ovis and A. marginale in Hungary

Recurring and spontaneously curing spring haemoglobinuria was recently reported in a small sheep flock in a selenium deficient area of northern Hungary. In blood smears of two animals showing clinical signs, Anaplasma-like inclusion bodies were seen in erythrocytes. To extend the scope of the study, 156 sheep from 5 flocks and 26 cattle from 9 farms in the region were examined serologically with a competitive ELISA to detect antibodies to Anaplasma marginale, A. centrale and A. ovis. The seropositivity in sheep was 99.4%, and in cattle 80.8%. A. ovis and A. marginale were identified by PCR and sequence analysis of the major surface protein (msp) 4 gene in sheep and cattle, respectively. Haemoglobinuria, an unusual clinical sign for anaplasmosis might have been a consequence of transient intravascular haemolysis facilitated by selenium deficiency in recently infected sheep, as indicated by the reduction of mean corpuscular haemoglobin concentration (MCHC). Membrane damage was also demonstrated for parenchymal cells, since their enzymes showed pronounced elevation in the plasma. Ticks collected from animals in the affected as well as in neighbouring flocks revealed the presence of Dermacentor marginatus, Ixodes ricinus and D. reticulatus, with the dominance of the first. The present data extend the northern latitude in the geographical occurrence of ovine anaplasmosis in Europe and reveal the endemicity of A. ovis and A. marginale in Hungary.     

Chemical trail-following by flying insects: a mechanism for orientation to a distant odor source

An aerial trail of odorous pheromone molecules extends downwind from a female pink bollworm moth that is receptive for mating. Males apparently sense the boundaries of the trail during their characteristic zigzag flights across it. Contrary to previous beliefs, the mechanism by which the males steer toward the odor source does not require a sensing of wind direction.     

Effect of added barley chromosomes on the flowering time of new wheat/winter barley addition lines in various environments

The heading characters and morphological traits of two partial sets of wheat–barley disomic addition lines, namely Mv9kr1/Igri and Asakaze/Manas, were evaluated under controlled environmental conditions in a phytotron under long-day, short-day and non-vernalised conditions and in field-sown experiments. The winter barley chromosome additions significantly influenced the flowering time of wheat both in the controlled environment test and under field-sown conditions. Of all the barley addition lines, the effect of the 4H and 7H additions was the most characteristic. The 7H addition lines were the earliest in both cultivar combinations in each treatment. In the Mv9kr1/Igri combination the 4H addition was the latest under all the environmental conditions. In the Asakaze/Manas combination 4H addition was the latest under short-day and long-day illumination in the phytotron but the 6H addition was the latest without vernalisation and in the field in 2012. There was 12 and 11 days difference between the flowering times of the 7H and 4H Mv9kr1/Igri and Asakaze/Manas addition lines in the field in 2012, which increased to 52 and 44 days under short-day illumination in the phytotron. In the winter wheat background, the addition of 2H carrying the photoperiod sensitivity gene Ppd-H1 decreased the flowering time under the short photoperiod regime, but had a very strong delaying effect under field-grown conditions. Considering the yield components under field conditions, 4H was the most fertile of the addition lines, while 7H showed the highest tillering capacity, and Igri 3H had good tillering capacity and the highest number of seeds per plant.     

<Emphasis Type="Italic">Helianthus tuberosus</Emphasis> L. agg. in the Carpathian Basin: a blessing or a curse?

Helianthus tuberosus agg., an herbaceous perennial plant from the Asteraceae family, is native to North America and was introduced as a crop to Europe in the seventeenth century. The status of this plant has been rather controversial in the Carpathian Basin. This paper reviews its taxonomy, chorology, and ethnobotany in this region, using data obtained from herbarium specimens, ethnobotanical surveys performed in Transylvania, and data published earlier, documenting the invasive features of the plant since the beginning of the nineteenth century. Our results suggest that (1) Helianthus tuberosus agg. has been present in the Carpathian Basin from the nineteenth century, (2) its taxonomy is problematic since the first part of the nineteenth century; (3) ethnobotanical surveys confirmed that the plant’s tuber has been locally used mostly as a food in Transylvania, collected from cultivation. Overall, Helianthus tuberosus agg. is both a ‘blessing and a curse’ at the same time, it is a valuable crop with beneficial nutritional value, and also an environmentally dangerous invasive element for the diversity of the flora of the Carpathian Basin.     

Survey on blood-sucking lice (Phthiraptera: Anoplura) of ruminants and pigs with molecular detection of Anaplasma and Rickettsia spp

Lice may serve as biological or mechanical vectors for various infectious agents. To investigate louse infestation of ruminants and pigs, and pathogens potentially transmitted by them, anopluran lice (n=1182) were collected in Hungary, and evaluated for the presence of anaplasma, rickettsia and haemotropic mycoplasma DNA. On cattle the following species were found: Linognathus vituli (57%), Haematopinus eurysternus (38%) and Solenopotes capillatus (5%). L. vituli had a lower mean individual count/host when compared to H. eurysternus. On calves only L. vituli was observed, with a higher louse burden than on full-grown cattle. H. eurysternus and S. capillatus were more likely to occur simultaneously with another species on the same host, than L. vituli. Goats infested with Linognathus stenopsis had the overall highest prevalence (68%), while pigs harbouring Haematopinus suis showed the lowest (<1%). Anaplasma DNA was detected in 50% of pools analysed. In L. vituli Anaplasma ovis (or a closely related novel Anaplasma marginale genotype) was identified. Anaplasma-positivity of H. suis suggests that pigs may extend the reservoir and/or host spectrum of relevant species. Anaplasma-infected L. stenopsis pools show for the first time that caprine anaplasmosis is endemic in Hungary. Rickettsia spp. were demonstrated from Linognathus spp. and H. eurysternus. No haemotropic mycoplasmas were detected in any samples. In conclusion, this is the first molecularly confirmed report of bovine and ovine Anaplasma spp. in L. vituli, L. stenopsis and H. suis. The present results suggest that phthirapterosis of domestic animals deserves more attention, and lice should be evaluated among the broad range of potential vectors of arthropod-borne pathogens.     

Molecular investigation of hard ticks (Acari: Ixodidae) and fleas (Siphonaptera: Pulicidae) as potential vectors of rickettsial and mycoplasmal agents

The aim of the present study was twofold. First, in general, to reveal new aspects of the potential vector role of ixodid ticks and fleas by screening large numbers of specimens with recently developed molecular biological methods. Second, to evaluate the occurrence of vector-borne infectious agents in a geographical context. Altogether 3442 unfed hard ticks (Ixodes ricinus, Dermacentor marginatus, D. reticulatus, Haemaphysalis inermis, H. concinna, H. punctata) and 939 fleas of cats and dogs (Ctenocephalides felis, C. canis, Pulex irritans) were collected in Hungary. DNA was extracted and analyzed in pools for representatives of the orders Rickettsiales and Mycoplasmatales.H. inermis was newly identified as the most important potential vector for Rickettsia helvetica in the study region. A novel Rickettsia genotype (designated ‘Candidatus R. hungarica’) was also detected in the same tick species, with a maximum of 95.8% gltA gene sequence identity to known rickettsiae. In addition, P. irritans tested positive for Rickettsia sp. RF2125, which has not been previously described in Europe. The human pathogen R. felis and the feline pathogen ‘Candidatus Mycoplasma turicensis’ were shown for the first time to occur in Central-Eastern Europe. Further novel findings include the presence of Spiroplasma spp. in D. marginatus and fleas. In conclusion, this molecular study extends the geographic range and vector spectrum of several arthropod-borne agents, some of which have zoonotic potential.     

Quantitative assessment of the morphology of the pig''s head used as a model in surgical experimentation. Part 1: Methods of Measurements.

Thirty-two surface measurements were described for assessment of the effect of complex surgical operations on the skeleton of the face in pigs. The methods of measurements imitate those of anthropometry. The surface measurements can complement cephalometry with data about the changes in the soft tissue and thus improve the documentation of the effect of surgery. This paper can help in the evaluation of complicated osteotomy procedures using the pig as the animal model, for facial reconstruction research in humans.     

The comparison of two different extenders for the improvement of large‐scale sperm cryopreservation in common carp (Cyprinus carpio)

In our study, a traditionally used (Grayling, already used in cyprinid species) and a newly tested (Pike) extender was tested to avoid sperm agglutination phenomenon following thawing during carp sperm cryopreservation. A large‐scale (elevated volume of sperm) freezing method in a controlled‐rate freezer using 5 ml straw and 10 ml cryotube was also systematically established. In all experiments, the sperm cryopreserved in using Grayling extender (except only one sample) showed an agglutination phenomenon (damaged and intact cells adhered to each other) after thawing where Pike extender resulted the regular cell suspension. No significant difference was observed between the two cryopreserved groups (Pike and Grayling extender) in all motility parameters using the 0.5 ml straw and the polystyrene box. Similarly, motility parameters did not show a significant difference in the two frozen groups with the 5 ml straw, also in the polystyrene box. A significantly higher progressive motility (pMOT, Grayling: 54% ± 8%, Pike: 37% ± 5%), straight line velocity (VSL, Grayling: 50 ± 5 µm/s, Pike: 39 ± 4 µm/s) and beat cross frequency (BCF, Grayling: 20 ± 1 Hz, Pike: 17 ± 1 Hz) was observed in the case of the grayling extender by the 5 ml straw cryopreserved in a controlled‐rate freezer (CRF) compare to the pike extender. A significantly higher VSL (Grayling: 45 ± 3 µm/s, Pike: 38 ± 4 µm/s) was observed by the grayling extender using the 10 ml cryotube than with the pike extender. Despite the randomly occurring differences in a few parameters, our new controlled freezing method using the newly tested Pike extender, the 5 ml straw or the 10 ml cryotube can be a good solution for the preservation of elevated volume of carp sperm.     

Adenovirus and mycoplasma infection in an ornate box turtle (Terrapene ornata ornata) in Hungary

A female, adult ornate box turtle (Terrapene ornata ornata) with fatty liver was submitted for virologic examination in Hungary. Signs of an adenovirus infection including degeneration of the liver cells, enlarged nuclei and intranuclear inclusion bodies were detected by light microscopic examination. The presence of an adenovirus was later confirmed by obtaining partial sequence data from the adenoviral DNA-dependent DNA-polymerase. Phylogenetic analyses revealed that this novel chelonian adenovirus was distinct from previously described reptilian adenoviruses, not belonging to any of the recognized genera of the family Adenoviridae. As a part of the routine diagnostic procedure for chelonians the detection of herpes-, rana- and iridoviruses together with Mycoplasma spp. was attempted. Amplicons were generated by a general mycoplasma polymerase chain reaction (PCR) targeting the 16S/23S ribosomal RNA (rRNA) intergenic spacer region, as well as, a specific Mycoplasma agassizii PCR targeting the 16S rRNA gene. Based on the analyses of partial sequences of the 16S rRNA gene, the Mycoplasma sp. of the ornate box turtle seemed to be identical with the recently described eastern box turtle (Terrapene carolina carolina) Mycoplasma sp. This is the first report of a novel chelonian adenovirus and a mycoplasma infection in an ornate box turtle (T. ornata ornata) in Europe.