1993 Annual Report Nebraska Game and Parks Commisison

Table of ContentsAdministration ... 2Budget and Fiscal ... 3Engineering ... 7Fisheries ... 10Information and Education ... 12 Law Enforcement ... 15Operations and Construction ... 17Outdoor Education ... 19Parks ... 20Planning and Programming ... 22Realty ... 23Resource Services ... 24Wildlife ... 2

The first cases of Dirofilaria repens infection in dogs in Lithuania

Biologijos katedraGamtos mokslų fakultetasLietuvos sveikatos mokslų universitetas. Veterinarijos akademijaUAB Siaurio šnaucerisVytauto Didžiojo universiteta

Tailoring a Soluble Diiron Monooxygenase for Synthesis of Aromatic <i>N</i>-oxides

The aromatic N-oxides have received increased attention over the last few years due to their potential application in medicine, agriculture and organic chemistry. As a green alternative in their synthesis, the biocatalytic method employing whole cells of Escherichia coli bearing phenol monooxygenase like protein PmlABCDEF (from here on &#8211; PML monooxygenase) has been introduced. In this work, site-directed mutagenesis was used to study the contributions of active site neighboring residues I106, A113, G109, F181, F200, F209 to the regiospecificity of N-oxidation. Based on chromogenic indole oxidation screening, a collection of PML mutants with altered catalytic properties was created. Among the tested mutants, the A113G variant acquired the most distinguishable N-oxidations capacity. This new variant of PML was able to produce dioxides (quinoxaline-1,4-dioxide, 2,5-dimethylpyrazine-1,4-dioxide) and specific mono-N-oxides (2,3,5-trimethylpyrazine-1-oxide) that were unachievable using the wild type PML. This mutant also featured reshaped regioselectivity as N-oxidation shifted towards quinazoline-1-oxide compared to quinazoline-3-oxide that is produced by the wild type PML

Prevalence and intensity of Sarcocystis spp. infection in animals slaughtered for food in Lithuania

The exact prevalence of Sarcocystis spp. infection in animals slaughtered for food is unknown in Lithuania. Therefore, the present study was initiated to evaluate Sarcocystis spp. infection in the carcasses of cattle (n = 206), sheep (n = 61), pigs (n = 73) and horses (n = 72) raised in Lithuania for food. The prevalence and intensity of Sarcocystis spp. infection were assessed under light microscopy by analysing 1 g of stained and squashed muscle samples. All the investigated muscle types (oesophagus, diaphragm, heart, neck, jaw, back, leg and tongue) were found to have been infected with microcysts rather than with macrocysts. A high prevalence of infection was established in cattle (44.9–98.1%) and sheep (100%), whereas the prevalence of this infection in pigs (30.1–50.0%) and horses (34.7–63.9%) was considered to be moderate. Significant differences in the infection prevalence were detected in the majority of muscle groups of cattle and in some muscle groups of pigs and horses. Similarly, significant differences in the median (Md) intensity of infection were observed in the majority of the muscle groups of cattle (Md = 4–29) and sheep (Md = 21–73) and only in some muscle groups of pigs (Md = 4.5–16) and horses (Md = 1–3). Cases of intense infection (> 40 cysts in a sample) were relatively often detected in sheep (44.9%) and cattle (19.1%), and rarely in pigs (3.7%). Hence, based on the varying rates of infection in the examined samples, the infection was identified as being intense in sheep and cattle, moderate in pigs and low in horses

Oxyfunctionalization of pyridine derivatives using whole cells of Burkholderia sp. MAK1

Pyridinols and pyridinamines are important intermediates with many applications in chemical industry. The pyridine derivatives are in great demand as synthons for pharmaceutical products. Moreover, pyridines are used either as biologically active substances or as building blocks for polymers with unique physical properties. Application of enzymes or whole cells is an attractive strategy for preparation of hydroxylated pyridines since the methods for chemical synthesis of pyridinols, particularly aminopyridinols, are usually limited or inefficient. Burkholderia sp. MAK1 (DSM102049), capable of using pyridin-2-ol as the sole carbon and energy source, was isolated from soil. Whole cells of Burkholderia sp. MAK1 were confirmed to possess a good ability to convert different pyridin- 2-amines and pyridin-2-ones into their 5-hydroxy derivatives. Moreover, several methylpyridines as well as methylated pyrazines were converted to appropriate N-oxides. In conclusion, regioselective oxyfunctionalization of pyridine derivatives using whole cells of Burkholderia sp. MAK1 is a promising method for the preparation of various pyridin-5-ols and pyridin-N-oxides

Biocatalytic synthesis of asymmetric water-soluble indirubin derivatives /

A method for the synthesis of asymmetric carboxy-substituted indirubins is presented. It employs indole-5-carboxylic acid or indole-6-carboxylic acid and 2-indolinone derivatives as substrates for bacterial monooxygenase-driven enzymatic bioconversion in different bacterial hosts. This bioconversion system achieved the highest titer of monocarboxyindirubin production of up to 327 mg L−1 for 5-bromoindirubin-6′-carboxylic acid during the 16-h incubation period. The purified monocarboxyindirubins exhibited high solubility in water, up to three orders of magnitude higher than that of indirubin. In addition, several monocarboxyindirubins, namely 1-methylindirubin-5′-carboxylic acid, possess potent antiproliferative activity against different cancer cell lines. Therefore, the synthesis method for monocarboxyindirubins described herein is an efficient and environmentally friendly bioconversion system and the synthesized monocarboxyindirubins show great promise due to their high water solubility and potential antiproliferative activity

Detection and molecular characterization of canine babesiosis causative agent Babesia canis in the naturally infected dog in Lithuania

Canine babesiosis caused by Babesia canis is an emerging infectious disease in Europe. Although previously uncommon, canine babesiosis has become quite frequent in Lithuania during the past decade. In the last few years an increasing number of cases with a wide variety of clinical signs have been recorded throughout the country. In Lithuania the identification of the disease agent in veterinarian clinics is based on a microscopic analysis of size and morphology. To date, no data on the genetic characterization of Babesia species in dogs have been documented for Lithuania. A total of 123 blood samples from dogs showing clinical signs of babesiosis on the basis of veterinary examination were tested for the presence of babesial parasites. Babesia isolated from dogs were detected and characterized by nested-PCR and sequence analysis of a fragment of the 18S rRNA gene. Babesia parasites were detected in blood smears of 94 dogs (76.4%). The molecular analysis revealed the presence of B. canis in 108 dogs (87.8%). Two genotypes of B. canis were distinguished on the basis on two nucleotide (GA → AG) substitutions observed in 18S rRNA gene sequences. The results of the present study provide knowledge of the distribution of B. canis genotypes in dogs in Lithuania, and show the necessity to use a molecular analysis for an accurate diagnosis of canine babesiosisBiologijos katedraGamtos mokslų fakultetasLietuvos sveikatos mokslų universitetas. Veterinarijos akademijaUAB Siaurio šnaucerisVytauto Didžiojo universiteta

Molecular identification of four Sarcocystis species in cattle from Lithuania, including S. hominis, and development of a rapid molecular detection method

Background: Six Sarcocystis species are known to use cattle (Bos taurus) as the intermediate host, two of which, S. hominis and S. heydorni, are zoonotic. There is a need for a method that will enable rapid identification of the Sarcocystis species in cattle. Methods: The diaphragm muscles of 102 cattle from Lithuania were examined for the presence of Sarcocystis spp., using two different methods for species identification. Individual sarcocysts were isolated from squash preparations of the diaphragm muscle under the light microscope, followed by genetic characterisation of excised cysts using sequence analysis of the 18S rRNA (18S rRNA) and cytochrome c oxidase subunit I (cox1) genes. The same cattle muscle samples were digested and species-specific PCR analyses targeting cox1 were developed to identify the Sarcocystis isolates to the species level. Results: Under the light microscope, sarcocysts were detected in 87.3% of animals, and Sarcocystis infection was verified in all digested samples. Three species, namely S. cruzi (n = 20), S. bovifelis (n = 23) and S. hirsuta (n = 6), were identified by DNA sequence analysis of isolated sarcocysts. Based on sequence analysis of cox1, the level of genetic variability depended on Sarcocystis species and geographical location. Four Sarcocystis species, S. cruzi (96.1%), S. bovifelis (71.6%), S. hirsuta (30.4%) and S. hominis (13.7%), were confirmed in the digested samples. In individual samples, the most common finding was two species of Sarcocystis (44.1%), followed by three species (26.5%), a single species (24.5%) and four species (4.9%). Conclusions: Although examination of tissue preparations under the light microscrope did not detect any sarcocysts belonging to S. hominis, this species was identified in the digested samples subjected to a cox1-specific PCR analysis. These results demonstrate the need for effective molecular diagnosis techniques to detect Sarcocystis spp., which may be present at a lower preva

Modification of the AFM Sensor by a Precisely Regulated Air Stream to Increase Imaging Speed and Accuracy in the Contact Mode

Increasing the imaging rate of atomic force microscopy (AFM) without impairing of the imaging quality is a challenging task, since the increase in the scanning speed leads to a number of artifacts related to the limited mechanical bandwidth of the AFM components. One of these artifacts is the loss of contact between the probe tip and the sample. We propose to apply an additional nonlinear force on the upper surface of a cantilever, which will help to keep the tip and surface in contact. In practice, this force can be produced by the precisely regulated airflow. Such an improvement affects the AFM system dynamics, which were evaluated using a mathematical model that is presented in this paper. The model defines the relationships between the additional nonlinear force, the pressure of the applied air stream, and the initial air gap between the upper surface of the cantilever and the end of the air duct. It was found that the nonlinear force created by the stream of compressed air (aerodynamic force) prevents the contact loss caused by the high scanning speed or the higher surface roughness, thus maintaining stable contact between the probe and the surface. This improvement allows us to effectively increase the scanning speed by at least 10 times using a soft (spring constant of 0.2 N/m) cantilever by applying the air pressure of 40 Pa. If a stiff cantilever (spring constant of 40 N/m) is used, the potential of vertical deviation improvement is twice is large. This method is suitable for use with different types of AFM sensors and it can be implemented practically without essential changes in AFM sensor design.This article belongs to the Section Physical Sensor