Animal Blood Serum Research Articles

Biotransformation of fenbendazole in sheep after administration of fenbendazole solid dispersion prepared by mechanochemical technique with arabinogalactane

The purpose of the research is to study the biotransformation of fenbendazole in the body of sheep after administration of fenbendazole solid dispersion (FSD) prepared by mechanochemical technique with arabinogalactan. Materials and methods. The FSD at a dose of 2.0 mg/kg for the active substance was administered orally to sheep. Animal blood serum samples were studied by high performance liquid chromatography / tandem mass spectrometry (HPLC-TMS) to determine the concentration of fenbendazole (FBZ) and its sulfoxide and sulfone metabolites at 0, 1, 2, 4, 6, 8, 12, 24, 33, 48 and 72 hours after administered FSD and initial FBZ (substance). FBZ and its metabolite residual quantity in the organs and tissues of the sheep was determined at 1, 3, 6, 11, and 21 days after the drug administration. The prepared sample and validated method were described. Results and discussion. A significant difference was found in the metabolism, pharmacokinetics, and timing of the FBZ and its metabolite elimination after the base drug (FBZ substance) and FSD were administered to sheep in an equal dose of 2.0 mg/kg for the active substance. FBZ and its metabolites began to be detected in blood serum 2 hours after the FSD and 4 hours after the base FBZ. Pharmacokinetic parameters of FBZ and its metabolites characterize a higher drug concentration in the blood and a longer retention time in the circulation after the FSD as compared with the base drug parameters. The FBZ and its metabolite maximum concentration was found in the organs and tissues of the sheep that received the FSD on day 3 in the liver amount of 4862.3±296.2 ng/g of FBZ; 18243.5±486.1 ng/g of sulfoxide; and 2482.3±132.4 ng/g of sulfone; and tens of times lower concentration after the base FBZ on day 6. FBZ and its metabolites were not detected in the organs and tissues of the sheep on day 16 after the base FBZ, and on day 21 after the FSD.

Bio-reductive synthesis of silver nanoparticles, its antibacterial efficiency, and possible toxicity in common carp fish (Cyprinus carpio).

The biological synthesis of nanoparticles is an emerging field of study that seeks to synthesize nanoparticles using non-chemical mechanisms such as microorganisms, plants, and animal blood serum. Among these, plants have gained particular attention due to their ease of handling, availability, and ability to synthesize a wide range of nanoparticles. Therefore, the current study aimed to fabricate the silver nanoparticles (AgNPs) using Chinese medicinal plants (CMP) for their possible toxicity in common carp fish (Cyprinus carpio). For this purpose, CMP was dried, ground, and used as a bio-reductive agent. The fabricated AgNPs were characterized and a well dispersed AgNPs were obtained. Moreover, the C. carpio was exposed to the AgNPs for bioaccumulation and histological alterations. The obtained findings revealed that the AgNPs were mostly accumulated in the intestines followed by the gills, muscles, liver, and brain. The accumulated AgNPs caused histological alterations in gills and intestines at the highest concentration (0.08 mg/L). However, very less alterations were caused by the lowest concentration, especially in the intestine. In conclusion, further in-depth research is needed to determine the risks associated with the usage of nanoparticles to reveal their harmful impacts on fish and the aquatic environment. HIGHLIGHTS: The biological fabrication of AgNPs is considered eco-friendly. Chinese medicinal plants play a significant role in AgNPs synthesis. AgNPs have excellent antibacterial activity. AgNPs are bioaccumulated in various organs of fish.

CORRECTION OF DISORDERS IN THE DIGESTIVE TRACT OF RATS WITH CHRONIC COLESTASIS WITH THE LEQUIN AND MINERAL COMPLEX

Problem. Complete or partial cessation of the flow of bile into the duodenum during cholestasis not only impairs the absorption of micro- and macroelements, fats and fat-soluble vitamins in the intestine, but also disrupts the microbiocenosis in the mucous membranes of the digestive tract. Knowledge of the mechanisms of disorders in the digestive tract caused by a decrease in the outflow of bile is important for the development of effective pathogenetic therapy. The drug Lequin (lecithin and quercetin), as well as the natural sorbent Mineralol, can be considered as such a preventive agent. Aim. To investigate the condition of the mucous membranes of the digestive tract of rats against the background of chronic cholestasis and to evaluate the effectiveness of prevention with the Mineralol and Lequin complex. Methods. The research was carried out on rats, which were divided into three groups: 1 group – intact, 2 group – rats that were modeled chronic cholestasis, 3 group – rats that were injected with Lequin (500 mg/kg) and Mineral (1 g/kg) against the background of cholestasis). Cholestasis in rats was modeled by ligation of the common bile duct under thiopental anesthesia. The drugs were administered orally to the rats of the 3rd group daily in the morning on an empty stomach for 4 months. Biochemical studies were carried out in blood serum (alkaline phosphatase, alanine aminotransferase, elastase activity), liver (elastase, acid phosphatase activity and malondialdehyde content) and mucous membranes of the oral cavity, small andlarge intestine (elastase, acid phosphatase activity and malondialdehyde content). Statistical processing of the research results was carried out using the Student-Fisher method. The main results. The conducted studies established liver parenchymal damage and cholestasis phenomena in rats with ligation of the common bile duct, which was confirmed by a decrease in alanine aminotransferase activity by 30.8% against the background of an increase in elastase activity by 42.0% and alkaline phosphatase activity by 43.6% in animal blood serum. Chronic cholestasis in rats led to the development of inflammatory processes in the liver: an increase in the activity of elastase by 24.5%, the activity of acid phosphatase by 29.8%, the level of malondialdehyde by 36.8%, along with a decrease in the antitoxic function of the liver: an increase in the activity of urease 2.2 times. Chronic cholestasis in rats caused the development of inflammation in the mucous membranes of the digestive tract: an increase in elastase activity by 28.4–41.1%, acid phosphatase activity by 19.3–45.6%, and malondialdehyde content by 55.7–127.0%. Daily prophylactic use of a complex of Lequin and Mineral drugs in conditions of chronic cholestasis in rats prevented the destruction of hepatocytes, the development of cholestatic phenomena, improved the antitoxic function of the liver, effectively prevented inflammatory processes, and the activation of lipid peroxidation in the mucous membranes of the digestive tract of animals. Conclusions. Established disorders in the mucous membranes of the digestive tract of rats with cholestasis are primarily caused by a decrease in the outflow of bile, which has antimicrobial properties, as a result of which toxic bile acids accumulate in the liver parenchyma along with a violation of the antitoxic function of the liver. The preventive effectiveness of the drugs was manifested due to the hepatoprotective, antioxidant, anti-inflammatory and choleretic effects of lecithin and quercetin, which, in combination with the sorption properties of Mineralol, made it possible to support the antitoxic function of the liver and its metabolic functions within normal limits. The conducted studies can be the basis for the use of the proposed drugs in the clinic for the treatment and prevention of cholestasis.

Evaluation of the Efficiency of a Soft Dosage Form for the Treatment of Osteoarthritis: Influence on the Level of Pro-inflammatory Cytokines

Introduction. Osteoarthritis is now considered to be a slowly progressive inflammatory disease that completely affects the joint. An important role in the development of this pathology is played by inflammation of the synovial membrane and ligaments (synovitis), supplemented by constant mechanical stress. Normally, a balance of anti-inflammatory and pro-inflammatory mediators is observed in cartilage, however, under the influence of risk factors, this balance shifts towards the latter.Aim. Evaluation of the effect of a previously developed soft dosage form containing meloxicam, a purine derivative and an immunomodulator M on the level of pro-inflammatory cytokines: IL-1α, IL-1β, IL-6 and TNF-α in osteoarthritis.Materials and methods. The experiment included 25 animals, which were divided into 5 groups by simple randomization: 1 – test (gel 0.5 %), 2 –test (gel 1 %), 3 – reference (Amelotex®, gel 1 %), 4 – control (gel base); 5 – intact. Preclinical modeling of the pathology was carried out by combined injection of 0.1 ml of a mixture of complete Freund's adjuvant with a 10 % suspension of talc in isotonic sodium chloride solution in a ratio of 1 : 10 into the cavity of the hock (tarsal) joint of Brown Norvay Catholic Rats male rats. Enzyme immunoassay of animal blood serum on the 28th day of the experiment was performed using standard ELISA plate kits (Cloud-Clone Corp., USA). Statistical data processing was performed using GraphPad Prism 8.0.2 software (GraphPad Software Inc., USA), differences were considered statistically significant at p < 0.05.Results and discussion. The developed compositions contributed to a decrease in the level of pro-inflammatory cytokines (IL-1α, IL-6 and TNF-α) compared with the main and reference gel preparation. At the same time, differences were found between the effect observed from the use of the test agents (gel 0.5 %, gel 1 %) and the reference drug in terms of the effect on the level of IL-1α and TNF-α, which indicates a greater effectiveness of the selected combination of active substances, because, unlike the single-component gel Amelotex®, the compositions developed by us additionally included a purine derivative and an immunomodulator M. The data obtained are important from the point of view of understanding the mechanism of action of a soft dosage form.Conclusion. Based on the results of previous and present studies, it is assumed that the combined composition of the soft dosage form with a half (0.5 %) concentration of meloxicam is of greatest interest for clinical practice, since its use at a high level of effectiveness additionally reduces the likelihood of adverse reactions from the non-steroidal anti-inflammatory drug, which is important in the case of long-term therapy of osteoarthritis.

Study of the influence of individual components in the cultivation environment on P. aeruginosa and A. baumannii biofilm-forming activity

Annotation. P. aeruginosa, A. baumannii are the causative agents of severe wound infections, ventilator-associated pneumonia, catheter-associated septic conditions and other severe pathological processes. The ability of bacteria to form biofilms in affected tissues plays an important role in the pathogenesis of these diseases. Therefore, the purpose of our work is to study the influence of certain external factors on the ability of bacteria to form biofilms. The ability of bacteria to form biofilms in affected tissues plays an important role in the pathogenesis of these diseases. 5 clinical strains of P. aeruginosa and 5 strains of A. baumannii were used in this research. The ability to form biofilms was studied in 96-well polystyrene plates by the spectrophotometric method, which is based on the ability of the crystal violet dye to bind to the cells and matrix of the biofilm. Statistical processing of the obtained results was carried out using Excel Microsoft Office tables. The effect of the presence of certain sugars and nitrogen-containing compounds in the cultivation medium on the processes of biofilm formation by pseudomonads and acinetobacteria was investigated. As a result of the research, it was established that acinetobacteria show the ability to form films, unlike pseudomonads, under the condition of their cultivation in sterile distilled water. The introduction of certain carbohydrates or urea into the cultivation medium intensified the biofilm-forming properties of pseudomonads to a much greater extent, compared to acinetobacteria. According to the research results, it was established that the most active process of biofilm formation occurs in both types of non-fermenting Gram-negative bacteria when there are animal blood serum proteins in the culture medium. The formation of biofilms is one of the main strategies that increases the survival of bacteria in the environment, as well as in the human body. The ability of bacteria to form biofilms is, among other things, a factor in the pathogenicity and virulence of bacteria. So, in recent years, special attention has been paid to the ability of microorganisms to form biofilms, and to the study of factors affecting this process.

Quantitative analysis of minor and trace elements in blood serum of dairy cattle (Bos taurus) by WDXRF


 
 
 
 Assessment of heavy metal content in biological fluids is essential to determine human and animal health as a result of exposure to contaminated environments, food, and water. WDXRF is a non-destructive method that involves no chemical treatments. It is possible to perform fast chemical analysis without producing hazardous waste and uses a small sample volume (± 50 mL), which is useful to veterinary usage. Therefore, the aim of this work was to implement an alternative method for minor and trace elements in blood serum samples of dairy cattle (Bos taurus) using WDXRF. It was implemented for the determination of Al, Cr, Co, Fe, Cu, As, and Hg, and the method was evaluated by comparing WDXRF and ICP-OES results of a set of 14 blood serum samples of dairy cattle using principal component analysis (PCA). Results of WDXRF and ICP-OES were statistically equivalent to all analyzed elements, except for Fe and Al, which were still comparable. WDXRF proved to be a suitable alternative methodology for heavy metal detection in animal blood serum samples due to the low sample volume. The results of this study enable a new and fast method for veterinary internal medicine, which requires a technique that quickly detects animals with a high concentration of toxic elements to assure the correct protocols to prevent the animal's death.
 
 
 

PSVII-6 Effects of Hydroponically Sprouted Cereal Grains on Feedlot Performance, Blood Serum Chemistry, and Apparent Nutrient Digestibility in Beef Cattle

Abstract Recent technological advancements in the local chain agricultural technology sector have produced automated systems that overcome pitfalls of traditional forage production. With significantly altered nutrient composition when compared with typical forages, research investigating the effects of hydroponically sprouted grain (HG) on feedlot growth and performance is needed. Angus steers (n = 40) averaging 274 kilograms body weight were randomly assigned to one of four pens in replicated pen experimental feedlot setting. Treatment pens were balanced for incoming weight, age, and genetic background to ensure uniformity throughout the experiment. Dietary treatments were randomly assigned and delivered continuously for an eight-week comparison period; diets were balanced for metabolizable energy, fiber, and crude protein content with 15% inclusion of HG administered in the treatment group. Nutrient digestibility was assessed through weekly manure sampling. Daily rate of gain, feed conversion ratio, and cost of gain was assessed through monthly replicated (n = 3) weight evaluations. Blood serum chemistry was evaluated through individual animal blood serum samples collected from the lateral tail vein. Multiple variables measured indicated positive and statistically significantly (p < 0.05) changes resulting from the inclusion of HG (Table 1). On a percentage basis, daily rate of gain and dry matter intake improved 12% and 11%, respectively. Marginal trends (p < 0.20) in nutrient digestibility were noted and blood serum metrics associated with nutrient bioavailability and rumen health were positive. Results highlight the potential feeding value of HG and automated production systems as a novel agricultural technology solution.

Long-term humoral immunogenicity, safety and protective efficacy of inactivated vaccine against reindeer rabies.

The core element of the reindeer rabies eradication strategy is regular application of vaccines to obtain and uphold a vaccination coverage sufficient for the ceasing of rabies virus transmission. This article presents the results of reindeer humoral immunity intensity and duration study after the immunization with two form of inactivated rabies vaccines (adjuvanted liquid vaccine and non-adjuvanted lyophilized vaccine) based on the Shchelkovo-51 rabies virus strain. Efficiency of post-vaccine immunity was assessed by measuring the animal blood serum virus-neutralizing antibody level in a neutralization test. The study determined the efficient rabies vaccine injection dose as equal to 3 ml. A single dose of 3 ml of these vaccines induced stable production of specific neutralizing antibodies in reindeer as early as 7 day after administration, and by 30 days after immunization, it significantly exceeded the minimal threshold level accepted by OIE. Two doses of vaccines administration with an interval of 30 days are required to achieve a strong immunity with the rabies-specific virus-neutralizing antibody titer of more than 0.5 IU/ml for at least 2 years. Our data do not support the benefit of an adjuvanted vaccine for the prevention of rabies in reindeer.

Study of immunogenic properties of B. suis 245 vaccine strain on domestic reindeer

This article presents the results of the study of immunogenic properties of the museum vaccine strain Brucella suis 245, from the collection of museum strains of brucella at All-Russian Research Institute of Experimental Veterinary Medicine named after K.I. Scriabin and Y.R. Kovalenko, which is of particular interest in anti-epizootic efficacy in the system of prevention and control of brucellosis of reindeer, as it is a homologous strain. Currently, a vaccine from the heterogeneous strain B. abortus 82 is used to prevent reindeer brucellosis. The subcutaneous injection of the Brucella suis 245 vaccine causes a local and general reaction of the body, as in the case of heterogeneous strains, the use of the vaccine at the root of the tongue (per os) eliminates all side effects. In the first days after the injection, the culture of the vaccine strain B. suis 245 is well settled in the reindeer. The humoral immune response of the body of the reindeer was studied within 15, 30, 75, 310 days after immunization by testing animal blood serum on brucellosis in RBT, SAT, Complement fixation test and immunodiffusion test with O-PS antigen. Agglutinating antibodies, detected in SAT, were detected in all animals on the 15th day after immunization and their levels in groups were as following: 1gr. - 466.6; 2 gr. - 150.0; 3 gr. - 125.0; 4 gr. - 100.0; 5 gr. - 233.3; 6 gr. - 100.0 ME. A decrease in SAT below the diagnostic level was observed in oral vaccinated animals after 75 days; and in animals vaccinated subcutaneously at 10 billion. m.k. with vaccine from strain 19 - in 310 days. In the body of reindeer grafted under the subcutaneous method, complementary binding antibodies appear on the 15th day after the injection of the vaccine, reaching the maximum level by 30 and the minimum level by 310 days.

Influence of Physiological Donor of Nitric Oxide on Blood Serum Crystallostasis in Rats

The aim of the work was to study the effect of intraperitoneal administration of dinitrosyl iron complexes with glutathione ligands on the initiated crystallogenesis of rat blood serum. The experiment was performed on 70 male Wistar rats divided into 7 equal groups. The first group of animals was intact (without any manipulation). Rats included in the other groups were administered intraperitoneal injection of 1 ml. 0.9% sodium chloride solution daily for 10 days. At the same time, animals of the third to sixth groups were additionally added to the injected solution with dinitrosyl iron complexes with glutathione ligands (the concentration of the agent was 0.15; 0.30; 0.45 and 0.60 mM, respectively). The rats of the seventh group received 1 ml. an aqueous solution of glutathione (0.15 mM). The study of the effect of DNIC on the crystallogenic properties of rat blood serum was carried out by the method of comparative tezioraphy using a special system of criteria. The data obtained as a result of this study indicate the presence of modification of the crystallogenic (initiating) properties of rat blood serum during the course intraperitoneal administration of a physiological donor of nitric oxide to animals – DNIC, and this effect turns out to be dose-dependent and having an extremum in the region of 0.3-0.45 mM. Taking into account the fact that the crystallogenic properties are a reflection of the component composition and one of the physico-chemical parameters of the biological fluid, it can be assumed that the metabolic response of the body to the use of the compound is also dose-dependent. At the same time, relatively low doses of the agent have the most optimal effect on metabolic processes, stimulating the antioxidant potential of blood serum with a corresponding decrease in the intensity of lipid peroxidation processes, contributing to the optimization of energy metabolism and increasing the activity of detoxification enzyme systems. The results of the teziographic evaluation of animal blood serum samples after a course of injections of DNIC in various concentrations clearly indicate the activating effect of the compound with respect to the crystallogenic (initiating) properties of biological fluid, which is most pronounced when using the substance in 0.3- and 0.45-millimolar aqueous solutions.

Study of the resistance of cows with mastitis depending on the age of lactation

Relevance.The breeding stock and young cattle in the conditions of industrial technology of animal husbandry experience an increased antigenic load, which leads to disturbances in the immune status and the development of inflammatory processes manifested by endometritis and mastitis, diarrheal and respiratory syndromes. It was relevant to conduct a comprehensive assessment of the immunological reactivity of the animal’s body according to the state of resistance in cattle with clinical and subclinical mastitis, depending on the age of lactation. As a result of complex studies, the specific and nonspecific reactivity of the body of cattle in different periods of lactation was studied.Methods.The object of the study were cows of a black-and-white Holstein breed (n = 450). Groups of animals were formed according to the age of lactation:1st lactation, 2–3rd lactation, 3–4th lactation and for health reasons: clinically healthy animals, with subclinical mastitis and clinical mastitis. Milk samples were taken from cows to count the number of somatic cells, and blood to determine the quantitative content of IgG immunoglobulin and indicators of nonspecific resistance.Results.It was found that with mastitis in cows of different lactation ages, almost identical changes in the immune status were observed. Quantitative determination of IgG levels in animal blood serum is the basis for assessing the immunity of cattle. The immune status in clinically healthy animals was characterized by a higher level of the total amount of IgG immunoglobulin in the blood serum (amounted to 22.25 ± 0.70 mg/ml), whereas in cows with a subclinical form of mastitis, the level of IgG immunoglobulin decreased by 12.3%, with a clinical form of mastitis — by 17.2%. The results also showed that in cows of different lactations with subclinical and clinical mastitis, the bactericidal activity (by 10.8%) and the lysozyme activity of the blood increased significantly (by 8.22%). In cows with clinical mastitis — by 8.6% and 9.7% respectively, compared with the indicators in groups of healthy animals. Thus, mastitis, occurring in acute and subacute form, leads to a decrease in the resistance of the cows’ organism.

Long-term exposure\xa0to high-concentration silver nanoparticles induced toxicity, fatality, bioaccumulation, and histological alteration in fish (Cyprinus carpio)

BackgroundCurrently, nanotechnology and nanoparticles have quickly emerged and have gained the attention of scientists due to their massive applications in environmental sectors. However, these environmental applications of silver nanoparticles potentially cause serious effects on terrestrial and aquatic organisms. In the current study, freshwater fish C. carpio were exposed to blood-mediated silver nanoparticles for toxicity, mortality, bioaccumulation, and histological alterations. Silver nanoparticles were fabricated using animal blood serum and their toxic effect was studied against common carp fish at different concentrations levels (0.03, 0.06, and 0.09 mg/L).ResultsThe findings have revealed a little influence of blood-induced silver nanoparticles on fish behavior at the highest concentration (0.09 mg/L). However, bioaccumulation of blood-mediated silver nanoparticles was reported in different organs of fish. Maximum bioaccumulation of silver nanoparticles was reported in the liver, followed by the intestine, gills, and muscles. Furthermore, the findings have shown that the bioaccumulation of silver nanoparticles led to histopathological alterations; including damaged structure of gill tissue and have caused necrosis. It is summarized that histopathological alteration in gill and intestine mostly occurred by the highest concentration of blood-induced silver nanoparticles (0.09 mg/L).ConclusionThis study provides evidence of the silver nanoparticles influence on aquatic life; however, further systematic studies are crucial to access the effects of AgNPs on aquatic life.

Pharmacological Evaluation of Cannabis indica For Their Aphrodisiac Potential

Objective: The plant Cannabis indica is a very popular and well known plant across the globe since thousands of years. In India also it is even more popular as bhang or marijuna from ancient time. A quite descriptions of this plant is written in various ancient books like the granths, vedas, puranas, ayurvedic books, dravyaguna, etc.. Number of medicinal properties have been described for different parts of the plant like flower buds(Ganja), Leaf(Bhang) & Leaf wax(Charas). All the parts of this plant have potent and complex psychotropic properties along with some additional therapeutic actions on different body systems. Here we have fond some opportunities to evaluate herbal extracts of leaves for having aphrodisiac potential. Method: The aphrodisiac activity of Cannabis indicaleaf extracts (Pet.ether, methenol & aqueous) were evaluated in male albino rats. Extracts were administered at a dose of 150mg/kg body weight. At a interval of 0,7,14,21 & 28 day several sexual behavior parameters were determined. At the end serum testosterone and FSH level were also determined. Results: This study also contributes to the pharmacology of aphrodisiacs as the plant Cannabis indica leaf extracts has significant effects on sexual physiology. It has shown effects on sexual behaviors as well as significant change in gonadotropin hormone labels in animal blood serum. Conclusion: This study provides a strong experimental evidence for its aphrodisiac use.

Simultaneous extraction of antibiotic and estrogen from animal blood serum using aqueous two-phase systems as predictor of environmental impact.

Efficient analytical methods are required for optimizing dosage of veterinary antibiotics and hormones in order to reduce toxicity and antimicrobial resistance in the environment. Thus, the objective of this work was to develop a rapid and low-cost method for determination of hormone estradiol and antibiotic chlortetracycline in bovine and porcine blood serum by aqueous two-phase system (ATPS) extraction and capillary electrophoresis quantification. ATPS based on ionic liquid cholinium alaninate and citrate salt along with mixtures of protic and aprotic polar solvents were evaluated in terms of recovery of extraction (%R). The liquid-liquid equilibrium, phase diagrams, and tie lines are discussed. Antibiotic migrated to solvent-rich phase (R ≈ 89.0%) to all systems. Estradiol migrates to ionic liquid-rich phase; however, addition of 10% methanol changed partition to solvent-rich phase (R ≈ 89.7%). The method has high recovery and cleanliness, is cost-efficient, scalable, and hence is adequate for screening of antibiotics and hormones tested in animal blood serum for dosage optimization and to predict their environment.

Ways to Improve Quality Control of Culture Medium Used for Mycoplasma Detection

An urgent safety concern associated with biological products is contamination with mycoplasmas, which may originate from donor tissues and organs, virus harvests, culture medium components, trypsin, animal blood serum, as well as be transmitted by personnel involved in the manufacture of medicines. Currently, due to an increase in the range of biologicals available, there is a need for more sensitive and specific test methods. In the Russian practice, microbiological (culture-based) testing of finished pharmaceutical products for mycoplasma contamination is performed using complex culture media whose sensitivity depends on the quality of proteins, ingredients, and reagents used. Growth promotion properties of the media are determined according to the State Pharmacopoeia of the Russian Federation, 14th ed., using a single test strain — Mycoplasma arginini G230 (M. arginini G230 industry reference material). The aim of the study was to analyse current Russian and foreign requirements for the quality control of culture media that are used for mycoplasma detection, in order to update and improve the quality control procedure in Russia. It was demonstrated that a compelling advantage of the State Pharmacopoeia of the Russian Federation is the possibility of using a semi-liquid culture medium which does not require special aerobic or anaerobic incubation conditions and allows for quantification of mycoplasma colonies and determination of mycoplasma titre in culture medium while testing its growth promotion properties using reference М. arginini G230 test strain. The analysis revealed some differences in Russian and foreign requirements for quality evaluation of culture media. These differences were taken into account when developing recommendations for improvement of the Russian test procedure, i.e. enlarging the range of test strains used and development of respective reference standards.

Minerals in the Blood, Hair, and Faeces of the Critically Endangered Western Derby Eland Under Human Care in Two Wildlife Reserves in Senegal.

The widely used means of investigating animal mineral profiles are blood serum or plasma and internal organ tissues. The acquisition of these types of samples can be invasive and requires much effort. These factors become key obstacles in the case of rare and elusive species such as the Western Derby eland (Taurotragus derbianus derbianus, WDE), which is a critically endangered antelope with a current distribution limited to the Niokolo Koba National Park, and two wildlife reserves in Senegal. One of the solutions to this problem is to collect easily accessible samples, such as faeces or fur, which may provide valid information about animal mineral status. Our study focuses on determining the macroelement and microelement levels in animal blood serum, hair, and faeces, and analysing their correlations to evaluate whether hair and/or faeces can be used as a proxy for blood mineral levels. Samples were collected from 11 individual WDEs (6 males, 5 females) during translocations within two reserves. Correlations of mineral concentrations in the blood, hair, and faeces were not found except for Fe in the faeces, which was positively correlated with Fe in the hair (r = 0.64, P < 0.05) and blood (r = 0.69, P < 0.05). The lack of correlations among the different types of samples may be because of the low number of samples; hence, we recommend conducting further research with a broader dataset. Our findings, however, currently indicate that faeces and fur analyses cannot stand alone for the assessment of the mineral status and the determination of WDEs' potential mineral deficiencies.

Early Cytokine Response After Vaccination with Coxiella Burnetii Phase I in an Infected Herd of Dairy Cattle.

IntroductionCoxiella (C.) burnetii, the aetiological agent of Q fever, is able to modulate the macrophage/T-lymphocyte axis in an infected organism and impair synthesis of monokines and lymphokines.Material and MethodsThe purpose of this research was to determine the levels of the cytokines that play a key role in the response to C. burnetii antigens (IL-1β, IL-2, IL-6, IL-10, IFN-γ and TNF-α) in the serum of animals originating from an infected herd prior to vaccination (day 0) and at 1, 7, and 21 days afterwards.ResultsThe vaccination of animals did not affect the production of IL-6, IL-1β, or IL-2. The serum levels of these cytokines were too low to measure in most of the samples. The initial levels of TNFα, IFNγ, and IL-10 were higher in seropositive than in seronegative animals, although significant differences between seropositive shedders and seropositive nonshedders appeared only in the levels of IFNγ and IL-10. Additionally, the course of the post-vaccination response concerning these two cytokines was different among seronegative nonshedders, seropositive nonshedders, and seropositive shedders.ConclusionIt seems that analysis of the IFNγ and IL-10 concentrations in animal blood serum may have some practical value in an assessment of the health status of seropositive animals and post-vaccination response.

CHANGES IN AMINO ACID COMPOSITION OF BLOOD PROTEIN HYDROLYSATE DEPENDING ON THE RAW MATERIAL PREPARATION MATERIAL

The raw material for blood protein hydrolysate preparation is whole animal blood, its clots and other serum production wastes. The dependence of amino acid composition of blood protein hydrolysate on the season of the raw material preparation was studied. The research lasted three years. It was demonstrated that the amino acid composition changed depending on the season. The peak, as a rule, was during summer months when their amount increased by 1.2–2.3 times and during autumn and winter it went down by 1.2–1.4 times (the difference is considerable, р &lt; 0,05). The peak of glutamic and asparagine acid growth was in November when their amount was 1.4 times higher then during the previous months (р &lt; 0,01). The increase of alanine, asparagine, valine, lysine, methionine, histidine, proline, tyrosine, threonine, and phenylalanine by 1.3–1.8 times was observed in March (the difference is considerable, р &lt; 0,05). The amount of histidine, glycine, leucine, serine, and tryptophane in the beginning of spring was at the same level and the amount of arginine, asparagine, isoleucine in March decreased by 1.2–1.6 times (the difference is considerable, р &lt; 0,01). So, it was determined that the dynamics of BPH amino acid composition was directly associated with the seasonal dynamics of physiological and biochemical cattle blood values. It was noted that in case of considerable change in absolute amino acid parameters their relative amount, in general, remained constant.

INFLUENCE OF CHANGESIN AMINO ACID COMPOSITION OF BLOOD PROTEIN HYDROLYSATE ON THE PRODUCTIVITY OF ВНК-21/2-17 CELL LINEAGE AND AMOUNT OF FMDV IMMUNOGENIC COMPONENTS

The raw material for blood protein hydrolysate preparation is whole animal blood, its clots and other serum production wastes. The dependence of ВНК-21/2-17 cell lineage productivity on the season of hydrolysate raw material preparation was observed. Cell growth stimulated by blood protein hydrolysate from raw material prepared in summer months 1.2–1.3 times exceeded cell growth stimulated by blood protein hydrolysate prepared in autumn-winter period (differences are significant, p &lt; 0,001). The amount of FMDV immunogenic components from “summer” blood protein hydrolysate 1.3–1.6 exceeded the amount obtained during other seasons of the year (differences are significant, p &lt; 0,001). There was found a direct correlation between BHK-21/2-17 cell lineage productivity, concentration of FMDV immunogenic components and seasonal dynamics of the contents of the following amino acids composing blood protein hydrolysates: alanin, asparagine, valine, histidine, glycine, isoleucine, leucine, lysine, methionine, proline, serine, tyrosine, threonine, tryptophane, and phenylalanine. To increase BHK-21/2-17 cell lineage productivity and concentration of FMDV immunogenic components it was suggested to add some additional amino acids into the culture growth medium for suspension cell culture in autumn-summer period together with blood protein hydrolysate

Determination of testosterone esters and nortestosterone esters in animal blood serum by LC-MS/MS

ABSTRACTA sensitive and robust confirmatory method for determination of steroid esters in blood serum is essential for reliable monitoring of possible illegal use of steroid hormones as growth promoters in meat production. A previously used sample preparation methodology was improved. The procedure consists of protein precipitation and removal of phospholipids by dispersive SPE Supel™ QuE Z-Sep (Sigma-Aldrich) followed by clean-up on alumina column and LC-MS/MS measurement. The modified method has been validated according to Commission Decision 2002/657/EC. Validation parameters for determination of six testosterone esters and five nortestosterone esters in bovine and porcine blood serum are presented in this article. Decision limits for all analytes were observed in the range 10–20 pg mL−1. The method described is considerably robust for bovine and porcine serum analyses and can be applied both for screening and confirmatory determination in routine residue monitoring.