Spectinomycin Research Articles

RESEARCH REGARDING THE DEVELOPMENT OF UNICELLULAR ALGAE FROM PROTOTHECA GENUS ISOLATED FROM MASTITIC COWS

Eighteen strains of Prototheca zopfii, an unicellular alga of Chlorococales order, Chlorellaceae family, Prototheca genus were isolated from subclinical mastitis cows. In our study, the Prototheca was able to develop on liquid and solid glucosed media, blood agar, potato agar, glucosed media with different pH values (from 2 to 9), media with antibiotics (lincomycin and spectinomycin). The Prototheca shows a good development in aerobic conditions at 37oC and the culture appears to be visible after 36 to 48 hours. The liquid media appears to be clear, but at the bottom of the culture tubes a granular sediment easy homogenisable is formed. At the surface a thin coating could appear. On solid media the colonies have small size, irregular ice crystal shape, but after 5 to 7 days at laboratory temperature the colonies grow to 3-4 mm, have matte whitish color, increased height and their surface has a blackberry or cauliflower appearance, aspects which are characteristic for Prototheca. On blood agar the colonies present the same aspect, without hemolysis. It is recommended to examine culture plates under a stereo microscope (x1.6 or x4). Acid and antibiotic based media have selective effect, and will eliminate bacteria and yeast growth.

THE ANTIBIOTIC RESISTANCE PHENOMENON IN CITROBACTER STRAINS ISOLATED FROM ANIMALS

The therapeutic choice of an antibiotic have to take into account, firstly, the ethiological agent, an also the susceptibility of this pathogen to antibiotics. Microorganisms from Micrococcus genus were isolated from animals and were identified using bacterioscopic and bacteriologic techniques, then were investigated through agar diffusion method about the susceptibility to different groups of antibiotics. The results of this study were interpreted according to the international standards and compared to the work of other researchers. Susceptibility of 100 % was registered for spectinomycin and tetracicline. It was found 66,66 % percentage of the bacterial strains to be sensitive to amoxicillin and 50 % to ampicillin, penicillin and only 33,33 % to potentiated sulfonamide. But the highest percentage of antibacterial resistance (100 %) was observed for flumequine and neomycin, followed by trimethoprim/sulphamethoxazole – 66,66 %, erytromycin – 60 % (in addition, 40 % intremediary strains) and penicillin (in addition, 50 % intermediary strains), ampicillin, kanamycin – 50 % . Although there were no reports of resistance to amoxicillin for the Micrococcus genus in this study, 33,33 % of the strains registered intermediary values for the inhibition areas around the microcomprimates, and 66 % of the bacterial strains presented susceptibility to this antibiotic. In conclusion, we recommend spectinomycin and tetracycline as the first choice for the Micrococcus genus strains, followed by penicillins, according to the results of this study.

Susceptibility of bacterial pathogens against lincomycin/spectinomycin (1/2), penicillin G/neomycin (1/1), and penicillin G/dihydrostreptomycin (1/1) as determined in the BfT-GermVet monitoring program 2004-2006.

A total of 368 bacterial pathogens, including 72 coagulase-positive and coagulase-variable staphylococci, 97 beta-haemolytic streptococci, 51 Escherichia coli, 75 Pasteurella multocida, 25 Mannheimia haemolytica, 25 Pseudomonas aeruginosa, and 23 Arcanobacterium pyogenes, were investigated for their susceptibility to the three combinations of antimicrobial agents lincomycin/spectinomycin (1/2), penicillin G/neomycin (1/1), and penicillin G/dihydrostreptomycin (1/1) in comparison to their susceptibility to the corresponding single substances. When comparing the minimum inhibitory concentrations (MICs) determined for any of the three combinations with those for the single substances, the lowest MIC of one of the two substances usually determined the MIC of the combination.This observation was made for all three combinations and all bacterial pathogens tested.Thus, it is assumed that the combination of lincomycin with spectinomycin as well as that of penicillin with either neomycin or dihydrostreptomycin resulted in an extended spectrum of target bacterial pathogens rather than in an increase in antimicrobial efficacy.

Discontinuation of Spectinomycin

Discontinuation of Spectinomycin

Determination of spectinomycin hydrochloride and its related substances by HPLC–ELSD and HPLC–MSn

A new and simple high-performance liquid chromatography-evaporative light scattering detection (HPLC-ELSD) method for the determination of spectinomycin hydrochloride and its related substances was developed. The column was Agilent SB-C(18) (250 mm x 4.6 mm, 5 microm). The mobile phase was 25 mM trifluoroacetic acid. The drift tube temperature was 40 degrees C. The pressure of nebulizing gas was 3.5 bar. Good separation of spectinomycin from main related substances could be achieved. The standard curve was rectilinear in the range of 0.07-3.8 mg/ml (r = 0.9997). Precision was 1.0% (R.S.D.). The limit of detection was 6 microg/ml. The method is simple and rapid, and the results are accurate and reproducible. The HPLC-MS(n) method was used to characterize the structures of impurities contained in the spectinomycin. In positive mode, impurities were elucidated by use of electrospray ion trap mass spectrometry in the multi-stage MS full scan mode. The possible structures of impurities C and D in spectinomycin were deduced based on the HPLC-MS(n) data.

Analysis of Spectinomycin by HPLC with Evaporative Light-Scattering Detection

A high-performance liquid chromatographic method with evaporative light-scattering detection (ELSD) has been developed for analysis of spectinomycin and related impurities. Separation of spectinomycin from structurally related impurities was achieved on a C18 column. The optimized mobile phase was 25 mmol L−1 ammonium acetate (pH 7.5)-methanol, 90:10 (v/v), at a flow rate of 0.6 mL min−1. The temperature of the drift tube of the ELSD was 95°C and the flow rate of carrier gas was 2.2 L min−1. The accuracy, specificity, precision, linearity, sensitivity, and robustness of the method were validated in accordance with ICH guidelines. In addition to determination of spectinomycin and related impurities, the method is also ideal for determination of the salts spectinomycin hydrochloride and spectinomycin sulfate.

Multiple doses of cefodizime are necessary for the treatment of Neisseria gonorrhoeae pharyngeal infection

A single dose of cefodizime (CDZM), ceftriaxone (CTRX), or spectinomycin (SPCM) is recommended for the treatment of gonococcal urethritis or uterine cervicitis in the era of multidrug-resistant Neisseria gonorrhoeae; namely, cefozopran-resistant N. gonorrhoeae (CZRNG). N. gonorrhoeae pharyngeal infection is not so rare in Japan; however, the proper treatment regimen for this infection is not clear. We previously found that a single dose of CDZM completely eradicated multidrug-resistant N. gonorrhoeae in patients with urethritis and uterine cervicitis, so we tried a single 1.0-g dose of CDZM for the treatment of N. gonorrhoeae pharyngeal infection, including infections with CZRNG. The eradication rate of N. gonorrhoeae from the pharynx was 63.0% with a single 1.0-g dose of CDZM, while the rate for CZRNG with the same dose of CDZM was 38.5%. N. gonorrhoeae was completely eradicated from the pharynx when patients received one or two additional doses of CDZM. Therefore, we concluded that two to three doses of CDZM were necessary for the treatment of N. gonorrhoeae pharyngeal infection including infection with CZRNG.

Expression of a Bacillus thuringiensis cry1C gene in plastid confers high insecticidal efficacy against tobacco cutworm - a Spodoptera insect

We have demonstrated that the toxic domain of a wild-type Bacillus thuringiensis (Bt) cry1 C gene could be expressed efficiently in the chloroplasts of transplastomic tobacco plants. resulting in high resistance to tobacco insects A DNA fragment with the toxic domain of a wild-type Bt cry1C gene was constructed under the control of rbcL promoter and an E colt thra terminator in a plasmid vector flanked by chloroplast sequences This plasmid vector xas then delivered into the chloroplast through biolistic bombardment technique Plant transformants that possessed spectinomycin and streptomycin resistance were selected, and the integration of cry1C gene in these transformants was confirmed by PCR and Southern analyses Results from PCR and Southern hybridization indicated that the truncated crylC gene was integrated into the chloroplast gene was we expected through homologous recombination High expression level of the integrated Bt crylC gene in the chloroplasts was observed through Northern and Western hybridization assays These transplastomic tobacco plants are highly toxic to the tobacco cutworm Spodoptera laura, causing a mortality rate of 76.9% to 100% after 72 h of feeding The truncated Bt toxin is expressed in high levels in the chloroplast (about l% of total proteins) and the plastid transgenes were known not to transmit through pollen So that, this study will facilitate not only improvement in breeding for insect-resistant plants. But also the prevention of contamination of transgenes among crop plants.

Residue analysis of spectinomycin in tissues of chicken and swine by HPLC

A reversed-phase HPLC method with ultraviolet detection using p-nitrophenyl hydrazine as a pre-column derivatizing reagent was investigated for the determination of the antibiotic spectinomycin (SPCM) in muscle, liver, kidney and fat of chicken and swine. SPCM was extracted from samples with 10% trichloroacetic acid saturated with EDTA-2Na, and then cleaned up with coupled Sep-Pak Plus PS-2 cartridges. The detection limit of SPCM was 0.02 microgram (potency)/g. Recoveries of SPCM ranged from 77.4 to 97.4% for chicken tissues and from 74.5 to 91.8% for swine tissues. The present method was used for the analysis of chicken tissues after the 11th day of withdrawal (SPCM-medicated drinking water: 500 mg (potency)/L, for 7 days), and swine tissues after the 14th day of withdrawal (SPCM-medicated feed: 100 mg (potency)/kg, for 7 days). Results showed that SPCM concentrations were lower than the MRLs in all tissues.

Screening of Salmonella isolates from a turkey production facility for antibiotic resistance

An ecological survey was conducted from April 1997 to June 1999 on four turkey flocks (F1 to F4). Turkey cecal contents, litter, waterers, feed, feeders, and environmental swabs were analyzed. Presence of Salmonella was determined using conventional microbiological screening techniques and confirmed by serology. Positive isolates were serotyped and screened for antibiotic resistance. From a total of 69 Salmonella isolates 25% were resistant to one or more antibiotics including gentamicin (G), spectinomycin (SP), streptomycin (S), tetracycline (T), tobramycin (TO), and trimethoprim/sulfamethoxazole. Isolates included 45 S. heidelberg, 13 S. senftenberg, 7 S. muenster, 2 S. anatum, and 2 S. worthington. Resistance to antibiotic(s) was highest among waterer isolates (55%) followed by environmental swabs (43%), feeder content samples (33%), turkey cecal contents (26%), and litter samples (5%). Frequencies of antibiotic-resistant Salmonella in F1, F2, and F4 were 27, 13, and 40%, respectively. Salmonella was undetected in F3. In F1, S. heidelberg from cecal content and waterer samples was resistant to G, SP, S, and T, whereas S. anatum from waterer samples was resistant to T and S. In F2, S. worthington from litter and feeder content samples was resistant to T, and in F4, S. muenster from environmental swabs was resistant to TO, S, SP, and G. Identifying preharvest sources and characterizing serotype and antibiotic-resistance profile can assist poultry producers and integrators in tracking movement of Salmonella on turkey farms.

Aktivitas Hormon Endogen dalam Buah Anggur Muscat OfAlexandria Muda tanpa Biji Hasill Induksi Antibiotika

The experiment was conducted to determine the activity of endogenous hormones in young seedless berry of Muscat of Alexandria (MOA) grapes induced by streptomycin (SM) and spectinomycin (SE). Young berries dipped in 200 ppm M or 200 ppm SE at 3 days before full-bloom and untreated berries were analyzed for the activity of . endogenous auxin, cytokinin. gibberellin and abscisic acid (ABA). The experiment was conducted at the Laboratory of ! Pomology, Faculty of Agriculture, Okayama University, Japan, from February 1999 to August 1999. Both SM and SE treatments did not affect ABA activities of young berries. Cytokinin activity were slightly decreased by SM and SE treatments at 6 days after treatment (DAT) and became more severe at J 0 and J 3 DAT The response of berry auxin activity to antibiotic treatment was similar but stronger than cytokinins. Antibiotic treatments significantly decreased the activity ofGA at 6 and 10 DAT 1 Keywords,' Antibiotics, Endogenous hormones, Spectinomycin, Streptomycin

Shortage of Spectinomycin

Shortage of Spectinomycin

Shortage of Spectinomycin

Shortage of Spectinomycin

Presence of 22-kDa protein reacting with sera in piglets experimentally infected with Brachyspira hyodysenteriae.

In order to examine the effect of spectinomycin on outbreaks of swine dysentery, experimental infection of piglets with Brachyspira hyodysenteriae was carried out. Feed with and without spectinomycin (SP) was given to each piglet ad libitum and the susceptibility of the piglets to infection with B. hyodysenteriae was compared between SP-treated and untreated piglets. The results showed that the SP-treated piglets did not display clinical signs of swine dysentery unlike the untreated piglets. The sera obtained from these piglets were examined by the microscopic agglutination test and antibodies to B. hyodysenteriae in both groups of experimentally infected piglets were detected and the reaction was serogroup-specific. The agglutination titers were very high in the untreated piglets with dysentery while the titers in the SP-treated piglets were lower than those in the untreated piglets. In addition, the immunoblotting technique was applied and the results demonstrated that 22- and 17-kDa proteins in strain ATCC 31212 (serogroup B) reacted strongly with the sera from the untreated piglets but not with the sera from the SP-treated piglets. The 22- and 17-kDa proteins also reacted with strain ATCC 27164 (serogroup A) which belongs to a different serogroup. The 22- and 17-kDa proteins were also confirmed in six other strains of B. hyodysenteriae which belong to six different serogroups. These proteins were sensitive to proteinase K. These results indicate that the 22- and 17-kDa proteins are common to eight strains of B. hyodysenteriae which differ serologically from each other.

Presence of 22-kDa protein reacting with sera in piglets experimentally infected with Brachyspira hyodysenteriae

In order to examine the effect of spectinomycin on outbreaks of swine dysentery, experimental infection of piglets with Brachyspira hyodysenteriae was carried out. Feed with and without spectinomycin (SP) was given to each piglet ad libitum and the susceptibility of the piglets to infection with B. hyodysenteriae was compared between SP-treated and untreated piglets. The results showed that the SP-treated piglets did not display clinical signs of swine dysentery unlike the untreated piglets. The sera obtained from these piglets were examined by the microscopic agglutination test and antibodies to B. hyodysenteriae in both groups of experimentally infected piglets were detected and the reaction was serogroup-specific. The agglutination titers were very high in the untreated piglets with dysentery while the titers in the SP-treated piglets were lower than those in the untreated piglets. In addition, the immunoblotting technique was applied and the results demonstrated that 22- and 17-kDa proteins in strain ATCC 31212 (serogroup B) reacted strongly with the sera from the untreated piglets but not with the sera from the SP-treated piglets. The 22- and 17-kDa proteins also reacted with strain ATCC 27164 (serogroup A) which belongs to a different serogroup. The 22- and 17-kDa proteins were also confirmed in six other strains of B. hyodysenteriae which belong to six different serogroups. These proteins were sensitive to proteinase K. These results indicate that the 22- and 17-kDa proteins are common to eight strains of B. hyodysenteriae which differ serologically from each other.

Antibiotik sebagai Induktor Buah Tanpa Biji pada Anggur

Four kinds of antibiotics were examined, viz. streptomycin (SM) and gentamicin (GM) as bactericidal antibiotica and tetracycline (TC) and spectinomycin (SE) as bacteriostatical antibiotics. Kyoho an Pione (tetraploid hybris cultivars) were used as the plant materials. Among the four antibiotics, SM and SE were effective to induce seedlessness in Muscat of Alexandria (MOA) and less effective for Kyoho and Pione. The seedless induction effect of SM and SE seemed not to be affected by their mode of action. Using the four cultivars, Kyoho, Pione, MOA, and Neo Muscat (NM), either SM or SE most effective to induce seedlessness when applied 3 days before full bloom. The ineffectiveness of antibiotics for Kyoho and Pione was indicated by the higher percentage of empty-seeded berries in these cultivars compared to MOA and Nm. Since there was no normal seed in the antibiotic-treated berries, this phenomenon indicated that the outer parts of the tetraploid ovules were more resistant to antibiotic treatments than those of diploid ovules. Key word: Grape, Seedless, Antibiotics

HPLC determination of residues of spectinomycin in various tissue types from husbandry animals†

An HPLC method was developed for the determination of bacteriostatic aminocyclitol spectinomycin (SP) in animal tissue products. These products included chicken eggs and edible fat, kidney, liver, muscle tissues from calf, poultry, pig and sheep. Residues of SP were extracted from homogenized tissue and egg-derived material with 25 mM citrate of pH 4.0, trichloroacetic acid and dichloromethane. The extract was purified and concentrated over a carboxylic acid-bonded solid-phase extraction (SPE) column. The SPE-eluate was analysed by cation-exchange HPLC involving a two-column switching system, post-column derivatization and fluorescence detection. Spectinomycin could be successfully determined at levels of 0.05 mg kg-1 and higher. Recoveries from spiked tissue material and from spiked egg material were in excess of 74% and did not show a concentration or tissue-type dependence. Precision of the elution position and signal response was better than 2%. Matrix effects and interference from lincomycin were less than 7 and 2%, respectively, on the signal response. Spectinomycin was shown to be stable at -20 degrees C in combined egg yolk and white over a test period of 12 weeks and in calf and sheep muscle tissue over a test period of 10 days. SP was, however, not stable at this temperature over a period of 12 months in chicken muscle tissue. Incurred SP residues were successfully determined in kidney and muscle tissue at the injection site of pigs administered with two doses of 15 mg kg-1 body weight SP with an intermittent withdrawal period of 15 days. Kidney showed higher concentrations and more persistent residues of SP than muscle tissue at the injection site.

Easy Enzyme-Linked Immunosorbent Assay for Spectinomycin in Chicken Plasma

An easy, sensitive, competitive indirect enzyme-linked immunosorbent assay (CI-ELISA) for spectinomycin in chicken plasma was developed. Preparation of a spectinomycin-bovine serum albumin conjugate in which the hapten is linked to the carrier protein through the C-4 position is described. Antibodies raised against antigens in rabbits had excellent specificity for spectinomycin, exhibiting a cross-reactivity of 44.0% with dihydrospectinomycin and 13.8% with tetrahydrospectinomycin. No cross-reactivity was observed with other antibiotics. The detection limit of the CI-ELISA was 2 ng/mL (equivalent into 40 ng/mL undiluted chicken plasma) spectinomycin. Known amounts (0.1-100 micrograms/mL) of spectinomycin were added to chicken plasma and then analyzed. Average recoveries were 97-110%. This procedure may be used without prior extraction of samples.

Residue Analysis of Spectinomycin in Chicken and Swine Tissues by HPLC

A reversed-phase HPLC method using p-nitrophenyl hydrazine as a pre-column derivatizing reagent was developed for the residue analysis of spectinomycin (SPCM) in chicken and swine tissues. SPCM was extracted from samples with 10% trichloroacetic acid saturated with EDTA-2Na, and then cleaned up with a Sep-pak tC18 cartridge. The detection limit of SPCM was 0.05μg (potency)/g. Recoveries of SPCM ranged from 73% to 95% for chicken tissues and from 75% to 87% for swine tissues. The present method was applied to the analysis of tissues of chicken and swine given various doses of SPCM.

Antibioresistance of Escherichia coli strains isolated in Morocco from chickens with colibacillosis

Two hundred and fifty eight isolates of Escherichia coli were made from autopsied chickens showing lesions of avian colibacillosis. Antibiograms showed high levels of resistance (greater than 40%) to sulphonamides (SSS), oxytetracycline (OT), trimethoprim+sulphamethoxazole (STX) and chloramphenicol (C). Medium frequencies of resistances (from 15 to 40%) were noted for streptomycin (S), spectinomycin (SPT), nalidixic acid (NA), oxolinic acid (OA), flumequine (UB) and enrofloxacine (ENR). For ampicillin (AM), gentamicin (GM), nitrofurans (FT), colistin (CS) and rifampin (RA) the frequencies of resistance were low (less than 15%). A linked resistance was observed for the 4 quinolones. A significant percentage of isolates (82.5%) were resistant to at least 2 antimicrobial agents. The most frequent antibiotypes were: C.OT.SSS.STX (4.65%), C.OT.SSS.STX.OA.NA.UB.ENR (4.65%), AM.S.C.OT.SSS.STX (4.26%) and OT.SSS.STX (3.87%).