In existing processes, the extraction of steviol glycosides from stevia leaves involves many process steps often including extraction by organic solvents. The purpose of the present study was to develop a process for the effective extraction of steviol glycosides, which can provide a concentrated juice exhibiting a high level of recovery with regard to the target compounds, rebaudioside A and stevioside. Pressurized Hot Water Extraction (PHWE) was first optimized with Response Surface Methodology in terms of maximized rebaudioside-A yield and minimized colour components. PHWE was then combined with pressing in a wine-press, resulting higher efficiency for extracting both steviol glycosides in comparison to the reported methods in the literature. Finally, spray drying was applied for both product stabilization and removal of contaminants.
In a survey of about 3000 dairy cows producing low somatic cell count (SCC) milk and kept on a large-scale dairy farm, California Mastitis Test (CMT) positivity was found in 2714 udder quarters of 1491 cows. Pathogenic microorganisms were isolated from 57.6% of these 2714 udder quarters during bacteriological examination. The commonest pathogens were coagulase-negative staphylococci (CNS, 41%) and Staphylococcus aureus (32.5%); however, udder infections caused by environmental streptococci (12.8%) and coliform bacteria (6.8%) were also common. All pathogens resulted in a significant increase of the SCC in individual bulk milk (IBM) samples. In the case of CNS, this SCC elevation in IBM was significantly lower than in the case of infection by the other pathogens. In spite of this, because of the high number of udder infections caused by CNS, the adverse effect exerted by CNS on dairy herds is considered to be substantial. It was found that 54.6% of all CMT-positive cows produced IBM of an SCC below 400 thousand per ml. The milk produced by 41% of the 315 cows excreting S. aureus also had an SCC below 400 thousand per ml. This poses a serious risk of infection to the healthy herdmates. At the same time, 11% of the infected cows produced IBM with an SCC below 100 thousand per ml. On the basis of these findings, only the regular analysis of SCC of IBM can be a reliable indicator of chronic intramammary infection. As the SCC of milk produced by CMT-positive cows (and especially of those excreting pathogens) tended to increase with advancing lactation, the authors suggest that an efficient drying-off therapy should be used to restore udder health and, whenever justified, culling of cows cannot be avoided either.
Because of the rapid development and spread of antimicrobial resistance it is important that a system be established to monitor antimicrobial resistance in pathogenic zoonotic and commensal bacteria of animal origin. Susceptibility testing of bacteria from carcasses and different samples of animal origin has been carried out in veterinary institutes for a long time but by an inconsistent methodology. The disc diffusion method proposed by the National Committee for Clinical Laboratory Standards (NCCLS) was introduced in all institutes in 1997. In order to obtain a coherent view of the antimicrobial resistance of bacteria a computer system was consulted, consisting of a central computer to store all data and some local computers attached to it through the network. At these local measuring stations computers are connected to a video camera, which displays the picture of Petri dishes on the monitor, and inhibition zone diameters of bacteria can be drawn with the mouse by the inspector. The software measures the diameters, evaluates whether or not the bacteria are sensitive, and stores the data. The evaluation is based upon the data of the NCCLS. The central computer can be connected to as many local computers with measuring stations as we wish, so it is suitable for an integrated system for monitoring trends in antimicrobial resistance of bacteria from animals, food and humans, facilitating comparison of the occurrence of resistance for each circumstance in the chain. It depends on the examiners which antibiotics they want to examine. Thirty-two different antibiotic panels were compiled, taking into consideration the active ingredients of medicinal products permitted for veterinary use in Hungary, natural resistance and cross-resistance, the mechanism of resistance and the animal species, i.e. which drugs were recommended for treatment in the given animal species, and the recommendations of the OIE Expert Group on Antimicrobial Resistance. The members of the panels can be changed any time, even during the measuring process. In addition to the inhibition zone diameters of bacteria the database also includes information about bacterial and animal species, the age of animals and the sample or organ where the bacteria are from. Since January 2001 the antibiotic susceptibility of E. coli, Salmonella, Campylobacter and Enterococcus strains isolated from the colons of slaughter cows, pigs and broiler chickens has also been examined. Each of the 19 counties of Hungary submits to the laboratory three tied colon samples from a herd of the above-mentioned animals every month.
Based on data of the Hungarian resistance monitoring system the antibiotic resistance of Staphylococcus strains of human and animal origin was studied. No methicillin-resistant staphylococci harbouring mecA gene were isolated from animals in 2001. Penicillin resistance, mediated by penicillinase production, was the most frequent among Staphylococcus aureus strains isolated from humans (96%), from bovine mastitis (55%), from foods (45%) and from dogs. In staphylococci isolated from animals low resistance percentages to aminoglycosides (0-2%), fluoroquinolones (0.5-3%) and sulphonamides (0.5-4%) were found but in strains isolated humans these figures were higher (1-14%, 5-18% and 3-31%, respectively). The most frequent antibiotic resistance profiles of strains isolated from animals and food were penicillin/tetracycline, penicillin/lincomycin and penicillin/lincomycin/tetracycline. Penicillin/tetracycline resistance was exhibited by strains from mastitis (3), samples from the meat industry (31), poultry flocks (1), poultry industry (1), noodle (1) and horses (2). Penicillin/lincomycin resistance was found in 10 Staphylococcus strains from mastitis, 1 from the dairy industry, 1 from the meat industry and 6 from dogs. Isolates from mastitis (2), from the dairy industry (2), from pigs (1), from the meat industry (1) and from poultry (1) harboured penicillin/lincomycin/tetracycline resistance pattern. Multiresistant strains were usually isolated only from one and sometimes from two animal species; therefore, the spread of defined resistant strains (clones) among different animal species could not be demonstrated. These results also suggest that the transfer of antibiotic resistance of S. aureus from animals to humans probably occurs less frequently than is generally assumed.
The objective of the investigations was to study the causes of abortion in sheep and goats in Hungary during a 7.5-year period. The authors investigated 246 cases of ovine and 75 cases of caprine abortions by different diagnostic methods. An infectious origin was found in 126 cases (51.2%) of ovine and 19 cases (25%) of caprine abortions. The most important cause of ovine and caprine abortions was Chlamydophila abortus infection with a prevalence of 46% and 17%, respectively. Other infections causing sheep and goat abortions were present only in 5.2% and 8% of the cases, respectively. The results obtained by different diagnostic methods are discussed.
The antibiotic resistance pattern of 1921 Staphylococcus strains isolated from animals and food within the last two years were examined using diffusion tests. Among them there were only 35 strains of S. aureus having an inhibition zone diameter of 15 mm or less, and 4 strains of coagulase-negative staphylococci (CNS) having a zone diameter of 18 mm or less to 1-µg oxacillin disk. These 39 strains were examined also by E-test to oxacillin and for the detection of the mecA gene by PCR in order to determine whether they might be real methicillin-resistant staphylococci. Among the 39 strains there were only two that were susceptible to penicillin by disk diffusion method; however, further examination by the penicillinase test showed that they produced ß-lactamase. While 19 (15 S. aureus, 4 CNS) strains were resistant and 7 strains were intermediate to oxacillin in disk diffusion test, the E-test gave 8 resistant and 5 intermediate results. Six out of the 8 oxacillin-resistant strains examined by disk diffusion and E-test harboured the mecA gene. Thus only 6 out of the examined 1921 strains proved to be mecA positive. These methicillin-resistant, mecA-positive strains (5 of the S. aureus strains and 1 of the S. epidermidis) originated from two dairy herds. The results prove that methicillin-resistant S. aureus (MRSA) strains in animals are really rare in Hungary. Eighteen strains were chosen and screened for minimal inhibitory concentration (MIC) of oxacillin with or without clavulanic acid or sulbactam, and three of them produced methicillinase enzyme.
The energy imbalance related predisposition to mastitis was studied in group-fed postpartum dairy cows (n = 333) kept in 4 large-scale units and producing milk of low somatic cell count (SCC). Blood samples were taken on Days 1-3 after calving for assaying some metabolites and hormones related to the negative energy balance (NEB). If mastitis was diagnosed later, aseptic milk samples were taken to identify the pathogens. Considering pathogen types [contagious pathogens: Staphylococcus (S.) aureus, Gram-positive (GP) environmental pathogens, and Gram-negative (GN) environmental pathogens + mastitis with no detectable pathogens (NDP)] separately, stepwise logistic regression was used to analyse the relation between the potential prognostic value of hormones and metabolites and mastitis outbreak. Only the elevated (= 1.00 mmol/l) serum ß-hydroxybutyrate (BHB) levels predisposed the cows to mastitis in the subsequent 4 weeks. This prognostic value of BHB was significant only in GN + NDP mastitis and in cases caused by GP environmental pathogens, but not in S. aureus mastitis (odds ratio: 5.333, 3.600 and 1.333, respectively).