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Thiourea‐based iminophosphorane (BIMP) organocatalysts featuring SPhos‐ or BIDIME phosphine units have been developed and successfully applied in the asymmetric addition of nitromethane to N‐Boc‐protected trifluoromethyl aryl ketimines. α‐Trifluoromethyl β‐nitroamines were obtained in 40–82% isolated yields and 80–95% enantioselectivities. A careful evaluation of the catalytic activity of BIMPs indicates that the catalysts derived from the combination via Staudinger reaction of a chiral 1,2‐amino alcohol‐derived thiourea‐organoazide with electron‐rich phosphines, promote the aza‐Henry reaction on fluorinated ketimines with the highest enantioselectivity, leading to the amine featuring a tetrasubstituted stereocenter in up to 95% ee. The reaction was performed also on gram scale, without loss of enantioselectivity. [ABSTRACT FROM AUTHOR]

The use of stereoselective phase-transfer catalysis as a reliable method for the enantioselective synthesis of optically active α-amino acid derivatives using achiral Schiff base esters has been well-developed in batch in the last 40 years. Recently, continuous flow technology has become of great interest in the academy and industry, since it offers safer process operating conditions and higher efficiency compared to a traditional batch processing. Herein, we wish to report the first example of enantioselective phase transfer benzylation of alanine Schiff base ester, under continuous flow conditions. Two different methodologies were investigated: a liquid-solid phase transfer catalytic benzylation using a packed-bed reactor and a liquid-liquid phase transfer catalytic benzylation in continuous stirred-tank reactors. Liquid-liquid phase transfer process in flow showed slightly better productivity than the batch process, while solid-liquid phase transfer benzylation proved much more advantageous in terms of productivity and space-time yield. Furthermore, continuous flow system allowed the isolation of benzylated product without any work up, with a significant simplification of the process. In both cases, phase transfer asymmetric benzylation promoted by Maruoka catalyst demonstrated high enantioselectivity of target quaternary amino ester in flow, up to 93% ee. [ABSTRACT FROM AUTHOR]

Background/Aim. Factors determining the risk of Lyme disease (LD) may be followed in terms of the type of habitat. The evaluation of the risk of Borrelia burgdorferi (B. burgdorferi) transmission in humans on Ixodes ricinus (I. ricinus) tick habitats is done by means of the ecological risk index, which determines the tick habitat, abundance and infection rate. The aim of this paper was to determine the value of ecological risk index (potential and actual risk – PR and AR) on green areas in 9 pаrks in Belgrade and establish the correlation of this index with tick bites in humans. Methods. Ticks were collected in parks by means of the flag hour method and examined for the presence of LD cause in dark-field microscopic analysis. Point values were assigned to certain parameters and potential and actual risk index evaluated for each habitat. The data on tick bites from the surveyed habitats were obtained from the Protocol of patients bitten by ticks of the Sector for Preventive Medicine, Institute of Epidemiology, Military Medical Academy in Belgrade. Analysis of variance (ANOVA), Tukey test and Pearson's coefficient were used in statistical analysis of data. Results. In Belgrade´s park habitats a high PR of B. burgdorferi transmission was determined, while AR for 4 habitats was categorised as PR, and limited for other 5 habitats. Statistically, in terms of AR values the following habitats were significantly different (p < 0.05): Hаjd Pаrk and Tаšmаjdаn; Hаjd Pаrk and Kаlemegdаn; Hаjd Pаrk and Pionirski Pаrk; Hаjd Pаrk and Bаnovo Brdo; Topčider and Tаšmаjdаn; Topčider and Kаlemegdаn; Topčider and Pionirski Pаrk; Topčider and Bаnovo Brdo Park. A statistically significant correlation (p < 0.05) between bites of adults and the number of bites of infected adults with the AR value was established. Conclusion. In parks of Belgrade, there is a limited AR of B. burgdorferi transmission on the average. The AR values vary from limited to the potential, depending on the ecological features of habitat, the number of collected ticks and their infection rate. In view of the correlation of AR with the bites in humans, this index is significant for assessing LD risk. [ABSTRACT FROM AUTHOR]

The aim of this study was to determine the influence of different feed composition and cultivation on the composition of fatty acids, their relationships, and the cholesterol content in the tissue of rainbow trout ( Oncorhynchus mykiss). Trout were reared in ponds at two different altitudes and raised on different feeds. The total amount of monounsaturated fatty acids in fillets of both groups showed no statistically significant difference. However, higher levels of n-6 fatty acids were found in Pond I trout fillets than in Pond II fillets, while Pond II fillets contained almost even amounts of n-3 and n-6 fatty acids. Total cholesterol content in Pond I trout fillets was significantly lower ( p < 0.05) than in Pond II fillets. The results indicate that the final composition of the trout fillets was mostly dependent on the feed the live fish had consumed; the altitude of the ponds had less effect. Also, both groups of trout could be used in the human diet as biologically valuable foods. Practical application: Our results show influence of altitude, feed, and fatty acid composition of feed on trout fillets. The average fatty acid level in the trout fillets increased in the order SFA

Background/Aim. The first case of human Lyme borreliosis (LB) in Serbia was recorded in 1987. The number of reported LB cases has increased in the past decade. The aim of this study was to estimate the density of Ixodes ricinus (I. ricinus) ticks, the prevalence of Borrelia burgdorferi sensu lato (B. burgdorferi) in them, and entomological risk index (ERI) at 19 Belgrade localities which were grouped into three categories (forests, park-forests, parks). The values of ERI were compared with the number of tick bites in humans. Methods. Ticks were collected monthly by using the flag hours method and the infection rate was determined by using dark field microscopy. The ERI value was calculated for each locality where the ticks were collected. The related data about tick bites was obtained from the patient protocol of the Institute of Epidemiology, Military Medical Academy, Belgrade. Results. The total number of collected ticks, the number of nymphs and the infection rates of the nymphs were significantly higher in forests (p < 0.05) than park-forests and parks. Statistically, the ERI value was significantly higher in forests than parks of Belgrade (2 = 7.78, p < 0.01). In March and July, the ERI value was also significantly higher in forests, than park-forests (p < 0.01) and parks (p < 0.01). May was the month with the highest ERI value in each ecological category (forests p < 0.05; park-forests p < 0.01; parks p < 0.001). However, the number of tick bites in humans did not correlate with ERI values. Conclusion. The obtained results indicate that the risk of tick bite and human exposure to B. burgdorferi sensu lato is present at all selected localities in Belgrade. For a more comprehensive Lyme disease risk assessment the method of entomological risk index assessment should be combined with other methods, taking into consideration all tick stages and the behaviour and habits of people who may get infected B. burgdorferi sensu lato. [ABSTRACT FROM AUTHOR]

Two new platinum(IV) complexes (1, trifluoperazinehydrochloride-aquapentachloridoplatinate(IV) and2, chlorpromazine-chlorpromazinehydrochloridepentachloridoplatinate(IV)) were synthesized in the reaction of K2[PtCl6] with trifluoperazine dihydrochloride (TF·2HCl) or chlorpromazine hydrochloride (CP·HCl). The complexes were characterized by elemental analysis, molar conductivity measurement, and spectral (IR,1H,13C, 2D1H–13C heteronuclear correlation spectra,195Pt NMR, and MS) methods. Outer-coordination sphere was proposed for1; while in2, the ligand was coordinated to the metal. The complexes exhibit antibacterial effect on strains ofBacillus subtilis,Bacillus cereus,Bacillus pumilus, and methicillin-resistantStaphylococcias Gram-positive bacteria and anEscherichia colias Gram-negative bacteria, as well as the reference strains. [ABSTRACT FROM PUBLISHER]

The thermal decomposition of [RuCl(η- p-cymene)] ( 1) and its biologically active N-alkylphenothiazine compounds of composition L[RuCl(η- p-cymene)] where L = CPH ( 2), TFH·HCl ( 3), and TRH ( 4) (chlorpromazine hydrochloride, CP·HCl; trifluoperazine dihydrochloride, TF·2HCl; and thioridazine hydrochloride, TR·HCl, respectively) has been studied. The crystal and molecular structure of compound 3 was determined earlier by single crystal X-ray diffraction analysis. The thermal data were collected by simultaneous TG/DSC measurements. For evolved gas detection, the qualitative reaction of chlorides with AgNO in an acidic solution was applied. The measurements were carried out in the temperature range to 700 °C in nitrogen atmosphere. Compounds of L[RuCl(η- p-cymene)] crystallize with water or water/2-propanole. On the basis of thermal data, the trend in the solvent bonding energies was assessed. [ABSTRACT FROM AUTHOR]

Abstract: Three new ruthenium(II) complexes 1–3 containing N-alkylphenothiazine molecules were synthesized by reaction of [RuCl2(η6-p-cymene)]2 with chlorpromazine hydrochloride (1), trifluoperazine dihydrochloride (2) or thioridazine hydrochloride (3). The compounds of the general formula L[RuCl3(η6-p-cymene)] were characterized by elemental analysis and spectroscopic methods (FT-IR, UV–Vis, 1H and 13C NMR). Complex 2 was structurally characterized by single crystal X-ray diffraction. In vitro cytotoxic activity of complexes 1–3 were assayed in four human carcinoma cell lines MCF-7, MDA-MB-453 (breast carcinoma), SW-480 (colon carcinoma) and IM9 (myeloma multiple cells). The highest cytotoxicity (12.1 ≤ IC50 ≤ 17.3 μM) and induced a total (SW-480) or almost total cell death (MCF-7, MDA-MB-453) at 25 μM in 48 h of treatment were observed for complex 2. The influence of three different doses (0.4, 4.5 and 90.4 μM/kg bw) of complex 2 on activities of antioxidants enzymes (superoxide dismutase (SOD) and catalase (CAT)) and lactate dehydrogenase (LDH) were investigated under physiological conditions. The effects on nitrite production () and level of erythrocytes malondialdehyde (MDA) in rats blood were evaluated, too. [Copyright &y& Elsevier]

Thermal decomposition of chlorpromazine hydrochloride (CP·HCl), trifluoperazine dihydrochloride (TF·2HCl) and thioridazine hydrochloride (TR·HCl), and the ruthenium complexes with dimethyl sulfoxide (dmso) of composition [RuCl(dmso)] and L[RuCl(dmso)]· xEtOH, L = CP·HCl, TF·2HCl or TR·HCl is described. The phenothiazines are stable to temperature range of 200-280 °C with an increasing stability order of TF·2HCl < CP·HCl < TR·HCl. The decomposition of all the compounds takes place in superposing steps. For detection of chlorides and sulfides, EGD analysis was performed. The decomposition pattern of the complexes, due to their similar structure, is similar. The thermal data unambiguously resolve the contradiction between the elemental analysis and X-ray structural data for (TF·2HCl)[RuCl(dmso)]Cl·EtOH. The compound crystallizes with one EtOH, evaporating in part at room temperature. [ABSTRACT FROM AUTHOR]

Abstract: Three new complexes of the general formula L[RuCl3(DMSO)3] (1–3), where L = chlorpromazine hydrochloride, trifluoroperazine dihydrochloride or thioridazine hydrochloride, were prepared and characterized by elemental analysis and spectroscopic methods (FT-IR, UV–Vis, 1H NMR and 13C NMR). In addition, the crystal structure of the complex 2 containing trifluoroperazine dihydrochloride was solved by single crystal X-ray diffraction. The complex crystallizes in the monoclinic system, space group P21/n, with a = 10.4935(7) Å, b = 18.6836(12) Å, c = 19.9250(13) Å, β = 98.448(2)°, V = 3864.0(4) Å3. The structure was refined to the agreement factors of R = 4.79%, R w = 11.23%. The effect of three different doses (0.4, 4.5 and 90.4 μM/kg bw) of complex 2 on superoxide dismutase (SOD) and catalase (CAT) activity was investigated under physiological conditions. Influence on nitrite production (NO2 −) and the level of erythrocytes malondialdehyde (MDA) in rats blood was also evaluated. Complex 2 did not affect the CAT enzyme activity in vivo and did not cause the hydroxyl radicals production. In the 0.4 and 4.5 μM/kg bw doses it showed almost the same or lower SOD activity and nitrite levels, while the dose of 90.4 μM/kg bw significantly increased these parameters. Finally, the cytotoxicity of complexes were assayed in four human carcinoma cell lines MCF-7, MDA-MB-453 (breast carcinoma), SW-480 (colon adenocarcinoma) and IM9 (myeloma multiple cells). Antiproliferative activity in vitro with low IC50 during 48 h of treatment was observed. [ABSTRACT FROM AUTHOR]

Background/Aim. Lyme disease (LD) is a cosmopolitan disease from the group of zoonoses that in humans can affect skins, joints, heart and/or nervous system. Research conducted in endemic areas, shows not only that the population, which lives or works in tick habitats, is in a high risk for getting infected by LD cause, but also that every environment itself is specific in regard to both vectors and B. burgdorferi species. In our environment, research of seropositivity to B. burgdorferi in persons exposed to vectors of LD have not been conducted. The aim of this study was to determine whether there was a higher risk for getting B. burgdorferi infection in the fieldworks of JKP "Zelenilo Beograd" than in other persons. Methods. Risk for getting LD in the fieldworkers of JKP "Zelenilo Beograd" (park-maintanance company) was determined according to seropositivity, the number and infection rate of LD vectors, and entomological risk index (ERI) value. Serum samples of 34 fieldworkers of JKP "Zelenilo Beograd", and 35 persons of the control group, were tested for the presence of specific IgM and IgG anti-B. burgdorferi antibodies by using ELISA test. In the research localities, the number of ticks was estimated by using the flag hours method, and tick infection rate was determined by dark field microscopy, with magnification rate of 400x. The data were analyzed statistically using χ² test, and Pearson's correlation analysis. Results. Out of 34 fieldworkes serum samples, 8 (23.5%) were positive on anti-B. burgdorferi antibody presence, while in the control group only one (2.9%) sample was positive (p < 0,05). Seropositivity rate varied from 0-66.6% among localities. Flag hours value among the localities ranged from 8.2-29.9, and tick infection rate ranged from 10.8-22.3%. ERI value was 0.03-0.15. Correlation between flag hour rate and seropositivity, tick infection rate and seropositivity, and also ERI and seropositivity was established. Conclusion. Frequency of B. burgdorferi seropositivity was significantly higher in green-surface maintenance workers, than in the control group not exposed to these habitats. A significant correlation was found between anti-B. burgdorferi antibodies findings and the number of ticks, tick infection rate, and ERI. The results obtained in our study indicate that there was a significantly higher risk for getting LD cause in the fieldworkers than in the control group. [ABSTRACT FROM AUTHOR]

This study was undertaken to investigate the effect of dietary fat supplementation with linseed oil and/or pig lard on performance, carcass characteristics, fatty acid profile and serum biochemistry parameters in broiler chickens. In the study, 240 one-day-old Cobb 500 broiler chickens were randomly distributed to one control and three experimental dietary groups: diet supplemented with pig lard; diet supplemented with linseed oil, and; diet supplemented with a 1:1 ratio of lard to linseed oil. The fat sources influenced serum biochemistry parameters and fatty acid profiles of drumsticks with thighs, but had no influence on growth performance or carcass characteristics. With dietary linseed oil, eicosapentaenoic acid and docosahexaenoic acid increased (P<0.05) in meat from drumsticks with thighs. Supplementation with pig lard significantly (P<0.05) increased the saturated and monounsaturated fatty acids in drumsticks with thighs. In conclusion, dietary incorporation of linseed oil and pig lard during starter, grower and finisher phases can enrich broiler chickens meat with n-3 PUFA. This study has clearly shown that linseed oil in broiler nutrition provided the best n-6/n-3 ratio. [ABSTRACT FROM AUTHOR]

Two non-steroidal antiinflammatory drugs indomethacin and mefenamic acid were coordinated to Ru(ii)–arenes to afford four new complexes. The cytotoxic activities of the ligands and ruthenium complexes were tested in three human cancer cell lines (K562, A549, MDA-MB-231) and non-tumour human fetal lung fibroblast cells (MRC-5) by MTT assay. Cytotoxicity studies revealed that indomethacin Ru(ii)–arene complexes 1 and 3 displayed good cytotoxicity and apparent cytoselective profiles. The IC50 values obtained in leukemia K562 cells were comparable to those of cisplatin (10.3 μM (CDDP), 11.9 μM (1) and 13.2 μM (3)). Flow cytometric analysis of 1 and 3 in triple-negative breast cancer MDA-MB-231 cells revealed an interesting mechanism of action. At IC50 concentrations, 1 and 3 arrested cell cycle progression in S phase and caused rapid accumulation of cells in sub-G1 phase (up to 48%), while Annexin V-FITC/PI staining showed simultaneous occurrence of apoptotic and necrotic cell populations at approximately similar levels of 20%. Measurement of reactive oxygen species (ROS) production by DCFH-DA staining confirmed the potential of 1 and 3 to increase ROS even more than cisplatin. The interaction of the complexes with serum albumins showed their potential ability to bind tightly and reversibly to albumins. The affinity of the complexes to calf-thymus DNA was investigated by UV-vis spectroscopy, viscosity measurements and fluorescence emission spectroscopy for competitive studies of the complexes with ethidium bromide, revealing that their interaction probably occurs via intercalation. Taken together, the results strongly suggest the potential of complexes 1 and 3 to alter cell cycle progression and cause DNA-damage by means of direct DNA-binding or indirectly by ROS production. [ABSTRACT FROM AUTHOR]

OBJECTIVE: The aim of our study is to show the potential and actual risk of transmission of Borrelia burgdorferi at a number of localities in Belgrade. MATERIALS & METHODS: On the territory of Belgrade on 16 sites of which 15 were test and belonged to different environmental categories (5 parks, 5 park-forests and 5 forests), while 1 was control, we investigated potential and actual risk of transmission of B. burgdorferi. From March to October 2012, the ticks were collected once a month by using the fl ag-hours method. Ticks were determined by the species and all Ixodes ricinus were viewed by dark fi eld microscopy. RESULTS: A total of 3158 ticks were collected, of which 2797 (88.6%) Ixodes ricinus, 175 (5.5%) Dermacentor reticulatus and 186 (5.9%) Rhipicephalus sanguineus. In forest were collected 1102 I. ricinus, in park-forests 730 and 569 in parks. Twelve sites met the requirement of high potential risk and of moderate there were three. In all forest sites, the actual risk in March and April was not certain and in the other months it was variable. In park-forests certain actual risk was registered 16 times during investigation, possible risk 23 and limited just once. In parks, certain actual risk was established 3 times, possible risk 30 and limited 7 times. Of 16 sites, there was one control (not chemicaly treated) where actual risk was registered during all months for 3 times. CONCLUSION: In Belgrade there are favorable conditions for maintaining potential and actual risk of transmission B. burgdorferi. [ABSTRACT FROM AUTHOR]

Mefenamic acid and indomethacin, medicaments belonging to the group of non-steroidal anti-inflammatory agents, were determined separately and simultaneously at anodically prepared BDDE. Differential pulse voltammetry was used to establish simple and fast analytical procedure based on electrooxidation of mefenamic acid at 0.6 V and indomethacin at 1.0 V (and 1.1 V). At optimal conditions, it was suitable for highly sensitive and selective determination of these compounds with LODs comparable to previously reported modified electrodes. Additionally, considering the importance of these compounds, we proposed precise and accurate quantification of these species based on green and simple pre-treatable electrochemical sensor. The effect of interfering agents (tablet ingredients and vitamins) was appeared to be negligibly confirming a favourable selectivity of the method. The proposed method was analytically applied by simultaneously determining mefenamic acid and indomethacin in pharmaceutical formulations. Unlabelled Image • Separate and simultaneous DPV determination of mefenamic acid and indomethacin at BDDE • Efficient procedure with simple electrochemical pretreatment at unmodified electrode • Applied for highly selective analysis of those compounds from pharmaceutical products [ABSTRACT FROM AUTHOR]

By WHO defi nition, the term biosecurity refer to the protection of microbial agents from loss, theft, diversion or intentional misuse.Security is not a new concept in biological research and medical laboratories. Biosafety and biosecurity are related, but not identical concepts. In some cases, biosecurity practices may confl ict with biosafety practices. Designing a biosecurity program that does not jeopardize laboratory operations or interfere with the conduct of research requires a familiarity with microbiology and the materials that require protection. Protecting pathogens and other sensitive biological materials while preserving the free exchange of research materials and information may present signifi cant institutional challenges. A risk management methodology can be used to identify the need for a biosecurity program. The risks need to be identifi ed, prioritized and resources allocated based on that prioritization. Management, researches and laboratory supervisors must be committed to being responsible stewards of infectious agents and toxins. Development of a biosecurity program should a collaborative process involving all stakeholders. The stakeholders include but are not limited to: senior management; scientifi c staff; human resource staff; and safety, security and engineering offi cials. The involvement of organizations and personnel responsible for a facility's overall security is critical because many potential biosecurity measures may already be in place as part of an existing safety or security program.This coordinated approach is critical in ensuring that the biosecurity program provides reasonable, timely and cost-effective solutions addressing the identifi ed security risks without unduly affecting the scientifi c or business enterprise or provision of clinical and diagnostic services. [ABSTRACT FROM AUTHOR]

Background/Aim. Despite numerous research of Lyme disease (LD), there are still many concerns about environmental of infectious agent of LD, as well as its prophylaxis, diagnosis and treatment. The aim of this work was to determine the risk of LD in relation to the way of removing ticks and duration of tick attachment. Methods. In the period from 2000 to 2007 a prospective study was conducted including persons with tick bite referred to the Institute of Epidemiology, Military Medical Academy, and followed for the occurrence of early Lyme disease up to six months after a tick bite. Epidemiological questionnaire was used to collect relevant information about the place and time of tick bites, the way of a removing tick, duration of tick attachment, remnants of a tick left in the skin (parts of the mouth device) and the signs of clinical manifestations of LD. Duration of tick attachment was determined on the basis of size of engorged tick and epidemiological data. Removed ticks were determined by the key of Pomerancev. Professional removing of attached tick was considered to be removing of tick with mechanical means by healthcare personnel. Fisher's exact test, Chi squares test and calculation of the relative risk (RR) were used for data analysis. Results. Of 3 126 patients with tick bite, clinical manifestations of LD were demonstrated in 19 (0.61%). In the group of subjects (n = 829) in which a tick was not removed professionally there were 17 (2.05%) cases with LD, while in the group of respondents (n = 2 297) in who a tick was removed professionally there were 2 (0.09%) cases with LD after tick bite (RR, 23.55; p < 0.0001). The disease was most frequent in the group of respondents with incompletely and unprofessionally removed ticks (2.46%). In the groups of patients with unprofessionally but completely removed ticks LD occurred in 0.89%, while in the group of subjects with a tick removed by an expert, but incompletely in 0.78% cases. The disease occurred rarely in the group with a tick removed completely and professionally (0.05%). There was no case of LD in the group of patients with a tick removed within 24 hours. The longer time of exposure after 24 hours, the higher absolute risk of disease was reported. Conclusion. In prevention of Lyme disease it is important to urgent remove a tick, to use a correct procedure of removing and to remove the whole tick without any remnants. [ABSTRACT FROM AUTHOR]

In the past decades there is growing awareness of the threat of bioterorrism which is defined as: «hostile use of microbes for political, religious and other ideological or criminal motivation». The risk of this threat is certainly augmented by the propulsion of scientific disciplines such as molecular biology and genetics, biotechnology, bioinformatics, and nanotechnology. Results and knowledge gained in these fields are typical dual-use products: from one side they serve for welfare of humankind, whereas from the other side they may be misused for deliberate impair of man and animal health as well as for the destruction of plant. In neutralizing bioterroristic act a major role play microbiology laboratory which principal goal is rapid detection and identification of biological agents. The main characteristics that determine laboratory abilities in managing these activities are existing levels of biosafety and biosecurity. Biosafety is the application of knowledge, techniques and equipment to prevent personal, laboratory and environmental exposure to potentially infectious agents or biohazards, i.e. biosafety defines the containment conditions under which infectious agents can be safely manipulated. Biosecurity is the effective implementation of measures to control access to pathogens and prevent unauthorized access. Vital elements that confere relevant level of biosafety and biosecurity are: laboratory equipment, staff competence,existing legislation, instructions and guidlines. If the use of biological weapon is suspected, basic principle during specimen processing is that screening and other tests should be performed in bisafety level 3 (BSL-3) rooms and cabinets, whereas in the case of probable use of highly contagious and lethal agents, laboratory tests should be carried out in the highest biocontainment level, BSL-4. Today definitive results are obtained in a significantly shorter period of time due to the expansion of molecular genetic methods, first of all PCR. Numerous other sensitive and specific methods are used with increasing frequency: immunodiagnostic (HHA in magnetic field, ELISA, ECORIGEN, TRF-Victor, Luminex- FC), multiple tests with high throughput, (Nanogen, Combimatrix, Affymetrix Resequencing), robotics (many sensors in one chip, high throughput), and biological mass spectrometry. Rationale protocol for the detection and identification of biological agents is as follow: 1. part of specimen, after inactivation (sterilization), is examined by (electron) microscopy; (result is obtained after 1-2h, preliminary); 2. part of specimen is seeded in nutrient media (liquid, enriched); after that PCR and/or immunological tests are performed (result after 6-12h, preliminary); 3. part of specimen is subjected to standard methods of isolation and identification (result after 2-5 days, definitive); 4. additional tests in animal model are carried out if virulence properties or other traits of agent in question are investigated; 5. part of specimen is saved (frozed) for additional investigation, if needed. It may be concluded that microbiological investigation play a key role in defending against bioterroristic attack. In dealing with this problem attention should be focused on: specimens handling, screening tests, speed, sensitivity and specificity of applied methods, reliability of obtained results , and on working conditions above all, taking into account existing levels of biosafety and biosecurity. [ABSTRACT FROM AUTHOR]