Methodological Aspects of Extraction, Phytochemical Characterization and Molecular Docking Studies of Salix caprea L. Bark and Leaves.

Contents of twelve selected bioactive substances and antioxidant potential of Salix caprea L. extracts were compared in its two vegetative organs (bark and leaves) and in terms of different ethanol/water mixtures used for extraction (30-70% aq, ethanol) and extraction time (30 min; 24, 48 and 72 h). The extracts were characterized by High Pressure Liquid Chromatography (HPLC), and total phenolics and flavonoids were determined spectrophotometrically. All secondary metabolites identified in Salix caprea L. extracts (gallic, chlorogenic and vanillic acid, epicatechin, rutin, quercetin and naringenin) were found more accumulated in bark. Salicin and p-hydroxybenzoic acid were detected in bark and ferulic, trans-cinnamic and p-coumaric acid in leaves extracts only. Rutin was most abundant bioactive compound both in bark (1.71 g/100 g of de) and leaves extracts (0.434 g/100 g of de). Bark extract with highest bioactive substances contents was obtained with 70% aq. ethanol as most suitable solvent during extraction time of 48 h. Molecular docking showed salicin to have similar affinity toward COX-2 as acetylsalicylic acid, but lower toward COX-1.

Chemical composition, antioxidant and antibacterial properties of Achillea collina Becker ex Heimerl s.l. and A. pannonica Scheele essential oils.

The in vitro antioxidant and antimicrobial activities of two Achillea millefolium (Adanson) Koch s.l species essential oils (A. collina Becker ex Heimerl s.l. and A. pannonica Scheele, Asteraceae) originating from the Golija and Radan mountains (Serbia) were investigated. The chemical profiles of the essential oils were evaluated by GC-MS. Antioxidant activity was assessed as free radical scavenging capacity (RSC) towards 2,2-diphenyl-1-picrylhydrazil (DPPH) radicals, together with effects on lipid peroxidation (LP). Antibacterial activity was examined on 21 bacterial strains. Based on the chemical composition of the essential oil, A. collina s.l. from Mount Golija was classified as a chamazulene chemotype (tetraploid). The high percentage of oxygenated monoterpenes and absence of azulene in the essential oil obtained from A. pannonica from Radan pointing that this population is octaploid. Essential oil of A. pannonica expressed stronger antimicrobial activity on almost all tested bacteria. Furthermore, this essential oil expressed higher scavenging effects on DPPH radical (IC(50) = 0.52 comparing to 0.62 mug/mL). Only in the LP evaluation, essential oil of A. collina s.l. from Golija exhibited stronger antioxidant activity (IC(50) = 0.75 comparing to 2.12 mug/mL).

New therapeutic potentials of milk thistle (Silybum marianum).

Silymarin is a bioflavonoid complex extract derived from dry seeds of Milk thistle [(Silybum marianum(L.) Gaemrnt. (Fam. Asteraceae/Compositaceae)] whose hepatoprotective effect has clinically been proved. Low toxicity, favorable pharmacokinetics, powerful antioxidant, detoxifying, preventive, protective and regenerative effects and side effects similar to placebo make silymarin extremely attractive and safe for therapeutic use. The medicinal properties of silymarin and its main component silibinin have been studied in the treatment of Alzheimer's disease, Parkinson's disease, sepsis, burns, osteoporosis, diabetes, cholestasis and hypercholesterolemia. Owing to its apoptotic effect, without cytotoxic effects, silymarin possesses potential applications in the treatment of various cancers. Silymarin is being examined as a neuro-, nephro- and cardio-protective in the damage of different etiologies due to its strong antioxidant potentials. Furthermore, it has fetoprotective (against the influence of alcohol) and prolactin effects and is safe to be used during pregnancy and lactation. Finally, the cosmetics industry is examining the antioxidant and UV-protective effects of silymarin. Further clinical studies and scientific evidence that silymarin and silibinin are effective in the therapy of various pathologies are indispensable in order to confirm their different flavonolignan pharmacological effects.

Influence of bile acids on rat gut microflora deterioration induced by oral ampicillin treatment

Background: Metabolic capacity of gut microflora is huge and this “microb” organ can be considered as second biggestmetabolic organ in body. The potential for an antibiotic to influence gut microflora is related to its spectrum of activity,pharmacokinetics, dosage and length of administration. In terms of pharmacokinetics, the rate of intestinal absorption playsa fundamental role. Apart from basic physiological functions, bile acids and their analogues are recognized as transportpromoters for other substances, in potentiating their action. The aim of this study was to demonstrate potential protectiveeffect of monoketocholic bile acid on rat intestinal microflora from oral ampicillin.Materials, Methods & Results: Eighteen Wistar rats were divided into three groups (n = 6). The experimental protocol wasapproved by Ethics Committee on Animal Use of the University Novi Sad. All animals received 10 mL/kg of body weightof drugs solutions per os by oral intubations. The animals have been treated twice daily for three days, with saline, ampicillin 500 mg/kg and ampicillin 500 mg/kg + monoketocholic bile acid (MKH) 4 mg/kg. The fecal pellets were collectedtwice, before and after the treatment was completed. Within 2 h of collection, samples of whole pellets were processedmicrobiologically. Weighed portions of feces were suspended 1:10 in sterile 0.9% NaCl and further diluted with samesolutions up to 1: 1013. The number of colony forming units (CFU) was determined by direct counting. Only the platescontaining 30 to 300 CFU were considered as valid. The ampicillin treated group, showed significant reduction...

Influence of bile acids on rat gut microflora deterioration induced by oral ampicillin treatment

Background: Metabolic capacity of gut microflora is huge and this “microb” organ can be considered as second biggestmetabolic organ in body. The potential for an antibiotic to influence gut microflora is related to its spectrum of activity,pharmacokinetics, dosage and length of administration. In terms of pharmacokinetics, the rate of intestinal absorption playsa fundamental role. Apart from basic physiological functions, bile acids and their analogues are recognized as transportpromoters for other substances, in potentiating their action. The aim of this study was to demonstrate potential protectiveeffect of monoketocholic bile acid on rat intestinal microflora from oral ampicillin.Materials, Methods & Results: Eighteen Wistar rats were divided into three groups (n = 6). The experimental protocol wasapproved by Ethics Committee on Animal Use of the University Novi Sad. All animals received 10 mL/kg of body weightof drugs solutions per os by oral intubations. The animals have been treated twice daily for three days, with saline, ampicillin 500 mg/kg and ampicillin 500 mg/kg + monoketocholic bile acid (MKH) 4 mg/kg. The fecal pellets were collectedtwice, before and after the treatment was completed. Within 2 h of collection, samples of whole pellets were processedmicrobiologically. Weighed portions of feces were suspended 1:10 in sterile 0.9% NaCl and further diluted with samesolutions up to 1: 1013. The number of colony forming units (CFU) was determined by direct counting. Only the platescontaining 30 to 300 CFU were considered as valid. The ampicillin treated group, showed significant reduction...(AU)

Seroepidemiological research of canine monocytic ehrlichiosis in the Autonomous Province of Vojvodina, Serbia / Seroepidemiological research of canine monocytic ehrlichiosis in the Autonomous Province of Vojvodina, Serbia

Background: Monocytic ehrlichiosis is an important disease in canine pathology and is present worldwide and has a potentially lethal outcome. This infection as a new infective disease of dogs and people and it has not been sufficiently researched, especially from the aspect of clinical picture, pathogenesis, diagnostics and differential diagnostics. The aims of this seroepidemiological research of the E. canis infection with dogs are to determine the values of G class antibodies prevalence to E. canis antigens, also to determine a agreement of positive serological findings between two immunodiagnostic tests (iELISA and IFAT), and to make an overview of the epidemiological characteristics of this canine infection. Materials, Methods & Results: One hundred dogs were involved in the seroepidemiological research of the E. canis infection. These dogs were from the City of Novi Sad area and its outskirts, Autonomous Province of Vojvodina, Serbia. The epidemiological survey of the closed type has been used in the research. The survey has been conducted with all of the dogs involved in this research. Apart from identifying veterinary practices and the patients (breed, sex, age), the survey was designed to collect data about the tick bites (first bite or a repeated one); the number of the removed ticks; the dogs lifestyles and information on whether the dogs had been abroad. An o

Background: Monocytic ehrlichiosis is an important disease in canine pathology and is present worldwide and has a potentially lethal outcome. This infection as a new infective disease of dogs and people and it has not been sufficiently researched, especially from the aspect of clinical picture, pathogenesis, diagnostics and differential diagnostics. The aims of this seroepidemiological research of the E. canis infection with dogs are to determine the values of G class antibodies prevalence to E. canis antigens, also to determine a agreement of positive serological findings between two immunodiagnostic tests (iELISA and IFAT), and to make an overview of the epidemiological characteristics of this canine infection. Materials, Methods & Results: One hundred dogs were involved in the seroepidemiological research of the E. canis infection. These dogs were from the City of Novi Sad area and its outskirts, Autonomous Province of Vojvodina, Serbia. The epidemiological survey of the closed type has been used in the research. The survey has been conducted with all of the dogs involved in this research. Apart from identifying veterinary practices and the patients (breed, sex, age), the survey was designed to collect data about the tick bites (first bite or a repeated one); the number of the removed ticks; the dogs lifestyles and information on whether the dogs had been abroad. An o

Antimicrobial activity of copper (II) complex with 1,2-bis [(1,3-diphenylpyrazol-4-yl)methyl] diaminoethane / Antimicrobial activity of copper (II) complex with 1,2-bis [(1,3-diphenylpyrazol-4-yl)methyl] diaminoethane

Background: Multi-resistant strains multiply daily, populate farms, hospitals and other ecological niches around the world, and cause serious infections in animals and humans, often leading to a fatal outcome. Researchers of all profiles are investigating intensively to fi nd new substances with antimicrobial activity. In the period between 1981 and 2002, 163 new chemical compounds were approved for use as drugs. Synthesized compounds have become much more interesting than the natural ones in the production of new antimicrobial agents. Some of these synthesized compounds are Copper (II) complex. The antimicrobial properties of copper were known in ancient Egypt (2000 BC), where it was used to sterilize water and wounds. Copper is still interesting for todays research. Materials, Methods & Results: Antimicrobial activity was tested using a microdilution method according to Clinical and Laboratory Standards Institute. The percentage of surviving bacteria was calculated in comparison to the number of bacteria placed in each well. Based on these results, using the Excel software package from Microsoft Office 2007, graphs were generated that showed the percentage of surviving bacteria depending on the corresponding effective concentrations of the tested substance. The function, which was used to approximate the experimental results, was determined using the Power Trendline supp

Background: Multi-resistant strains multiply daily, populate farms, hospitals and other ecological niches around the world, and cause serious infections in animals and humans, often leading to a fatal outcome. Researchers of all profiles are investigating intensively to fi nd new substances with antimicrobial activity. In the period between 1981 and 2002, 163 new chemical compounds were approved for use as drugs. Synthesized compounds have become much more interesting than the natural ones in the production of new antimicrobial agents. Some of these synthesized compounds are Copper (II) complex. The antimicrobial properties of copper were known in ancient Egypt (2000 BC), where it was used to sterilize water and wounds. Copper is still interesting for todays research. Materials, Methods & Results: Antimicrobial activity was tested using a microdilution method according to Clinical and Laboratory Standards Institute. The percentage of surviving bacteria was calculated in comparison to the number of bacteria placed in each well. Based on these results, using the Excel software package from Microsoft Office 2007, graphs were generated that showed the percentage of surviving bacteria depending on the corresponding effective concentrations of the tested substance. The function, which was used to approximate the experimental results, was determined using the Power Trendline supp

Antimicrobial activity of copper (ii) complex with 1, 2-bis [(1, 3-diphenylpyrazol-4-yl)methyl] diaminoethane

Background: Multi-resistant strains multiply daily, populate farms, hospitals and other ecological niches around the world, and cause serious infections in animals and humans, often leading to a fatal outcome. Researchers of all profiles are investigating intensively to find new substances with antimicrobial activity. In the period between 1981 and 2002, 163 new chemical compounds were approved for use as drugs. Synthesized compounds have become much more interesting than the natural ones in the production of new antimicrobial agents. Some of these synthesized compounds are Copper (II) complex. The antimicrobial properties of copper were known in ancient Egypt (2000 BC), where it was used to sterilize water and wounds. Copper is still interesting for today's research. Materials, Methods & Results: Antimicrobial activity was tested using a microdilution method according to Clinical and Laboratory Standards Institute. The percentage of surviving bacteria was calculated in comparison to the number of bacteria placed in each well. Based on these results, using the Excel software package from Microsoft Office 2007, graphs were generated that showed the percentage of surviving bacteria depending on the corresponding effective concentrations of the tested substance. The function, which was used to approximate the experimental results, was determined using the Power Trendline supplement from the Microsoft Excel program. Cytotoxicity (growth inhibition) was evaluated by tetrazolium colorimetric MTT assay, after exposure of cells to the tested compound for 48 h. Inhibition of growth was expressed as a percentage of cytotoxicity and calculated according to the following equation: (1-A test/A control) x 100. MBC99.9 and MIC99 of the test substance were lowest for Arcanobacterium haemolyticum being 0.2 mg/L and 0.0054 mg/L, respectively. The highest values were obtained for Arcanobacterium pyogenes and methicillin-resistant Staphylococcus aureus (MRSA) 488.002 mg/L and 20.2 mg/L. MIC80 for all four strains ranged from 0.00002 to 0.0023 mg/L. Measured values for MIC99 are 0.00545 mg/L for A. haemolyticum, 0.0443199 mg/L for R. equi, 0.0520712 mg/L for S. aureus and 2.36378 mg/L for A. pyogenes. Values for MIC99.9 ranged from 0.236134 to 488,002 mg/L. Most of the MIC values obtained in this study are significantly lower than those reported by other researchers. The values we obtained were lower as compared to MIC values for standard antibiotics, which were considered acceptable by the relevant institutions. This speaks in favor of a stronger antibacterial effect of our tested substances. In regards to cytotoxicity, the obtained MIC80 doses were lower than toxic, whereas MIC90 could be classified as low-toxic (less than 0.0625 µM), except of Arcanobacterium pyogenes only. According to the IC50 values, the compound Cu (L) Br2·MeOH was 6.4-fold and 4.8-fold more potent against HCT116 cells compared to normal lung fi broblasts and SW620 cells, respectively. Discussion: Copper (II) complex with an arylpyrazole ligand exhibits strong antibacterial properties, and it shows bacteriostatic effect at concentrations where there is no cytotoxic effect in normal human cells. The emergence of multi-resistant strains of pathogenic bacteria is a growing problem worldwide. Therefore, each new compound with potential antimicrobial activity, especially if it is not cytotoxic in effective dosage, deserves the attention of the scientific community. In this paper, we presented a newly synthesized substance with such properties.

Seroepidemiological research of canine monocytic ehrlichiosis in the autonomous province of Vojvodina, Serbia

Background: Monocytic ehrlichiosis is an important disease in canine pathology and is present worldwide and has a potentially lethal outcome. This infection as a 'new' infective disease of dogs and people and it has not been sufficiently researched, especially from the aspect of clinical picture, pathogenesis, diagnostics and differential diagnostics. The aims of this seroepidemiological research of the E. canis infection with dogs are to determine the values of G class antibodies prevalence to E. canis antigens, also to determine a agreement of positive serological findings between two immunodiagnostic tests (iELISA and IFAT), and to make an overview of the epidemiological characteristics of this canine infection. Materials, Methods & Results: One hundred dogs were involved in the seroepidemiological research of the E. canis infection. These dogs were from the City of Novi Sad area and its outskirts, Autonomous Province of Vojvodina, Serbia. The epidemiological survey of the closed type has been used in the research. The survey has been conducted with all of the dogs involved in this research. Apart from identifying veterinary practices and the patients (breed, sex, age), the survey was designed to collect data about the tick bites (first bite or a repeated one); the number of the removed ticks; the dogs' lifestyles and information on whether the dogs had been abroad. An overall of 62 ticks were collected from the bodies of these dogs. By applying the screening iELISA test, the seroprevalence of class G specific antibodies was determined with dogs to E. canis antigens in 25%, while 75% of dogs were negative to the presence of specific antibodies of IgG class to E. canis. Out of 25 seropositive dogs, with antibodies to E. canis discovered through iELISA test, 12 (48%) dogs were in contact with ticks for the first time, while the repeated contact with ticks was experienced by 9 (36%) dogs. For 4 (16%) dogs, the owners negated that their dog had been bitten by a tick. Upon the application of a confirming IFAT, a seroprevalence was determined of specific class G antibodies with dogs to E. canis antigens in 16%, while 84% were negative to the presence of specific antibodies of IgG class antibodies to E. canis. Out of 16 seropositive dogs, in which antibodies were discovered with IFAT to E. canis, 9 (56%) dogs had been in contact with ticks for the fi rst time, while 4 dogs (25%) had repeated contacts with ticks. For 3 dogs (19%), the owners negated that ticks had bitten their dogs. Through a statistical processing of the results gained from the applied immunodiagnostic tests, a moderate agreement (Kappa value of 0.48) was determined. In the group of dogs that had been bitten by a tick, three types were determined: Dermacentor marginatus, Rhipicephalus sanguineus and Ixodes ricinus. Overall 35 ticks of Dermacentor marginatus ticks were replaced from the dogs' bodies just as were 15 Rhipicephalus sanguineus ticks and 12 Ixodes ricinus ticks. Discussion: The determined values of the seroprevalence in our country (iELISA - 25% and IFAT - 16%) are in harmony with the acquired values in other countries with warm climate. The values of seroprevalence determined in this research point to the fact that this disease must be taken into consideration in everyday health protection of dogs, especially after a registered tick bite within a differential diagnostics procedure.

A proposal of a diagnostic protocol for isolation of Corynebacterium ulcerans from cow's milk

Background: Literature about presence Corynebacterium ulcerans in milk samples from cows with mastitis is rare and in the literature there are only a few reports. In this study the isolation and identification of Corynebacterium ulcerans from mastitis in dairy cows were done. Also, optimization of diagnostic protocols to identify Corynebacterium ulcerans was performed. Materials, Methods & Results: The investigation was performed at the cattle farm that is characterized by closed housing system diary Holstein-Friesian cows during an outbreak of acute mastitis. Milk samples from 298 lactating cows were collected in sterile sampling tubes. Before the collection of quarter milk samples, the udder was thoroughly cleaned with soap and water and rubbed to dry. All collected milk samples were examined for mastitis using California mastitis test, which was carried out by the method first described by Schalm and Noorlander. Equal volumes (5 mL) of commercial CMT reagent and quarter milk were mixed and the changes in milk fluidity and viscosity were observed. Sample portions (0.1 mL each) were inoculated on 10% sheep blood agar, Endo agar and Sabouraud agar as well as on thioglycolate medium and nutrient broth. Primary plates were incubated for 3 days at 37ºC in aerobic conditions. Cultural, morphological and conventional biochemical testing was done. The survey was complemented by double CAMP and plasma coagulation tube test. All 14 isolates developed a synergistic haemolysis with Rhodococcus equi (ATCC 6939) and inverse CAMP phenomenon with Staphylococcus aureus and coagulated rabbit plasma. Final diagnosis was confirmed using API Coryne V 2.0 and software program by BioMerieux1, revealing an identity rate of 99.9%, accuracy rate T = 1, test count = 0. Discussion: The first fourteen isolates of Corynebacterium ulcerans have been identified in our country, on the basis of a diagnostic protocol that is proposed in this paper. In our experience double CAMP test, rabbit plasma coagulation, catalase, oxidase tests and selected biochemical parameters, are sufficient as a diagnostic minimum. In the diagnostics of bacterial agents in cow mastitis, the attention of a bacteriologist is mostly limited to most widespread agents of mastitis, the isolation of which is mandatory pursuant to national legislation (Staphylococcus aureus and Streptococcus agalactiae). A more important reason for "missing" Corynebacterium ulcerans in the diagnosis is its colonial morphology that could resemble organisms of the genus Staphylococcus. Complex and expensive diagnostic procedure that is not available to most laboratories is also responsible for the small number of reports of isolation C. ulcerans. Furthermore, in routine work C. ulcerans could be misidentified with Staphylococcus intermedius, because of cultural similarity, positive plasma coagulation tube test and absence of manitol fermentation of both species. This paper is a report on isolation and identification of Corynebacterium ulcerans from milk of cows with mastitis, as well as a suggestion of a diagnostic protocol available for routine work in most veterinary microbiology laboratory. Therefore we suggest as the diagnostic protocol double CAMP test to be used as a complementary method to rabbit plasma coagulation tube test.

A proposal of a diagnostic protocol for isolation of Corynebacterium ulcerans from cows milk / A proposal of a diagnostic protocol for isolation of Corynebacterium ulcerans from cows milk

Background: Literature about presence Corynebacterium ulcerans in milk samples from cows with mastitis is rare and in the literature there are only a few reports. In this study the isolation and identification of Corynebacterium ulcerans from mastitis in dairy cows were done. Also, optimization of diagnostic protocols to identify Corynebacterium ulcerans was performed.Materials, Methods & Results: The investigation was performed at the cattle farm that is characterized by closed housing system diary Holstein-Friesian cows during an outbreak of acute mastitis. Milk samples from 298 lactating cows were collected in sterile sampling tubes. Before the collection of quarter milk samples, the udder was thoroughly cleaned with soap and water and rubbed to dry. All collected milk samples were examined for mastitis using California mastitis test, which was carried out by the method first described by Schalm and Noorlander. Equal volumes (5 mL) of commercial CMT reagent and quarter milk were mixed and the changes in milk fluidity and viscosity were observed. Sample portions (0.1 mL each) were inoculated on 10% sheep blood agar, Endo agar and Sabouraud agar as well as on thioglycolate medium and nutrient broth. Primary plates were incubated for 3 days at 37o C in aerobic conditions. Cultural, morphological and conventional biochemical testing was done. The survey was complemented by d

Background: Literature about presence Corynebacterium ulcerans in milk samples from cows with mastitis is rare and in the literature there are only a few reports. In this study the isolation and identification of Corynebacterium ulcerans from mastitis in dairy cows were done. Also, optimization of diagnostic protocols to identify Corynebacterium ulcerans was performed.Materials, Methods & Results: The investigation was performed at the cattle farm that is characterized by closed housing system diary Holstein-Friesian cows during an outbreak of acute mastitis. Milk samples from 298 lactating cows were collected in sterile sampling tubes. Before the collection of quarter milk samples, the udder was thoroughly cleaned with soap and water and rubbed to dry. All collected milk samples were examined for mastitis using California mastitis test, which was carried out by the method first described by Schalm and Noorlander. Equal volumes (5 mL) of commercial CMT reagent and quarter milk were mixed and the changes in milk fluidity and viscosity were observed. Sample portions (0.1 mL each) were inoculated on 10% sheep blood agar, Endo agar and Sabouraud agar as well as on thioglycolate medium and nutrient broth. Primary plates were incubated for 3 days at 37o C in aerobic conditions. Cultural, morphological and conventional biochemical testing was done. The survey was complemented by d

Antimicrobial activity of copper (II) complex with 1,2-bis [(1,3-diphenylpyrazol-4-yl)methyl] diaminoethane / Antimicrobial activity of copper (II) complex with 1,2-bis [(1,3-diphenylpyrazol-4-yl)methyl] diaminoethane

Background: Multi-resistant strains multiply daily, populate farms, hospitals and other ecological niches around the world, and cause serious infections in animals and humans, often leading to a fatal outcome. Researchers of all profiles are investigating intensively to fi nd new substances with antimicrobial activity. In the period between 1981 and 2002, 163 new chemical compounds were approved for use as drugs. Synthesized compounds have become much more interesting than the natural ones in the production of new antimicrobial agents. Some of these synthesized compounds are Copper (II) complex. The antimicrobial properties of copper were known in ancient Egypt (2000 BC), where it was used to sterilize water and wounds. Copper is still interesting for todays research. Materials, Methods & Results: Antimicrobial activity was tested using a microdilution method according to Clinical and Laboratory Standards Institute. The percentage of surviving bacteria was calculated in comparison to the number of bacteria placed in each well. Based on these results, using the Excel software package from Microsoft Office 2007, graphs were generated that showed the percentage of surviving bacteria depending on the corresponding effective concentrations of the tested substance. The function, which was used to approximate the experimental results, was determined using the Power Trendline supp

Background: Multi-resistant strains multiply daily, populate farms, hospitals and other ecological niches around the world, and cause serious infections in animals and humans, often leading to a fatal outcome. Researchers of all profiles are investigating intensively to fi nd new substances with antimicrobial activity. In the period between 1981 and 2002, 163 new chemical compounds were approved for use as drugs. Synthesized compounds have become much more interesting than the natural ones in the production of new antimicrobial agents. Some of these synthesized compounds are Copper (II) complex. The antimicrobial properties of copper were known in ancient Egypt (2000 BC), where it was used to sterilize water and wounds. Copper is still interesting for todays research. Materials, Methods & Results: Antimicrobial activity was tested using a microdilution method according to Clinical and Laboratory Standards Institute. The percentage of surviving bacteria was calculated in comparison to the number of bacteria placed in each well. Based on these results, using the Excel software package from Microsoft Office 2007, graphs were generated that showed the percentage of surviving bacteria depending on the corresponding effective concentrations of the tested substance. The function, which was used to approximate the experimental results, was determined using the Power Trendline supp

A proposal of a diagnostic protocol for isolation of Corynebacterium ulcerans from cows milk / A proposal of a diagnostic protocol for isolation of Corynebacterium ulcerans from cows milk

Background: Literature about presence Corynebacterium ulcerans in milk samples from cows with mastitis is rare and in the literature there are only a few reports. In this study the isolation and identification of Corynebacterium ulcerans from mastitis in dairy cows were done. Also, optimization of diagnostic protocols to identify Corynebacterium ulcerans was performed.Materials, Methods & Results: The investigation was performed at the cattle farm that is characterized by closed housing system diary Holstein-Friesian cows during an outbreak of acute mastitis. Milk samples from 298 lactating cows were collected in sterile sampling tubes. Before the collection of quarter milk samples, the udder was thoroughly cleaned with soap and water and rubbed to dry. All collected milk samples were examined for mastitis using California mastitis test, which was carried out by the method first described by Schalm and Noorlander. Equal volumes (5 mL) of commercial CMT reagent and quarter milk were mixed and the changes in milk fluidity and viscosity were observed. Sample portions (0.1 mL each) were inoculated on 10% sheep blood agar, Endo agar and Sabouraud agar as well as on thioglycolate medium and nutrient broth. Primary plates were incubated for 3 days at 37o C in aerobic conditions. Cultural, morphological and conventional biochemical testing was done. The survey was complemented by d

Background: Literature about presence Corynebacterium ulcerans in milk samples from cows with mastitis is rare and in the literature there are only a few reports. In this study the isolation and identification of Corynebacterium ulcerans from mastitis in dairy cows were done. Also, optimization of diagnostic protocols to identify Corynebacterium ulcerans was performed.Materials, Methods & Results: The investigation was performed at the cattle farm that is characterized by closed housing system diary Holstein-Friesian cows during an outbreak of acute mastitis. Milk samples from 298 lactating cows were collected in sterile sampling tubes. Before the collection of quarter milk samples, the udder was thoroughly cleaned with soap and water and rubbed to dry. All collected milk samples were examined for mastitis using California mastitis test, which was carried out by the method first described by Schalm and Noorlander. Equal volumes (5 mL) of commercial CMT reagent and quarter milk were mixed and the changes in milk fluidity and viscosity were observed. Sample portions (0.1 mL each) were inoculated on 10% sheep blood agar, Endo agar and Sabouraud agar as well as on thioglycolate medium and nutrient broth. Primary plates were incubated for 3 days at 37o C in aerobic conditions. Cultural, morphological and conventional biochemical testing was done. The survey was complemented by d

Seroepidemiological research of canine monocytic ehrlichiosis in the Autonomous Province of Vojvodina, Serbia / Seroepidemiological research of canine monocytic ehrlichiosis in the Autonomous Province of Vojvodina, Serbia

Background: Monocytic ehrlichiosis is an important disease in canine pathology and is present worldwide and has a potentially lethal outcome. This infection as a new infective disease of dogs and people and it has not been sufficiently researched, especially from the aspect of clinical picture, pathogenesis, diagnostics and differential diagnostics. The aims of this seroepidemiological research of the E. canis infection with dogs are to determine the values of G class antibodies prevalence to E. canis antigens, also to determine a agreement of positive serological findings between two immunodiagnostic tests (iELISA and IFAT), and to make an overview of the epidemiological characteristics of this canine infection. Materials, Methods & Results: One hundred dogs were involved in the seroepidemiological research of the E. canis infection. These dogs were from the City of Novi Sad area and its outskirts, Autonomous Province of Vojvodina, Serbia. The epidemiological survey of the closed type has been used in the research. The survey has been conducted with all of the dogs involved in this research. Apart from identifying veterinary practices and the patients (breed, sex, age), the survey was designed to collect data about the tick bites (first bite or a repeated one); the number of the removed ticks; the dogs lifestyles and information on whether the dogs had been abroad. An o

Background: Monocytic ehrlichiosis is an important disease in canine pathology and is present worldwide and has a potentially lethal outcome. This infection as a new infective disease of dogs and people and it has not been sufficiently researched, especially from the aspect of clinical picture, pathogenesis, diagnostics and differential diagnostics. The aims of this seroepidemiological research of the E. canis infection with dogs are to determine the values of G class antibodies prevalence to E. canis antigens, also to determine a agreement of positive serological findings between two immunodiagnostic tests (iELISA and IFAT), and to make an overview of the epidemiological characteristics of this canine infection. Materials, Methods & Results: One hundred dogs were involved in the seroepidemiological research of the E. canis infection. These dogs were from the City of Novi Sad area and its outskirts, Autonomous Province of Vojvodina, Serbia. The epidemiological survey of the closed type has been used in the research. The survey has been conducted with all of the dogs involved in this research. Apart from identifying veterinary practices and the patients (breed, sex, age), the survey was designed to collect data about the tick bites (first bite or a repeated one); the number of the removed ticks; the dogs lifestyles and information on whether the dogs had been abroad. An o