Cytotoxic effect of aminoglycoside antibiotics on the mammalian cell lines.

Aminoglycoside antibiotics have been used for treating serious but also routine infections in veterinary and human medicine for many years. The basic aim of this work is to evaluate the cytotoxicity of dihydrostreptomycin and neomycin in vitro on three cell cultures - BHK-21 (Syrian golden hamster kidney fibroblast), VERO (African green monkey kidney fibroblast) and FEA (feline embryonic fibroblast) cells. The morphological changes were examined by Giemsa staining. Cells were dried and visualized under fluorescence microscope. After the exposure to different experimental doses of dihydrostreptomycin (812.5-20000 µg/mL) and neomycin (1000-20000 µg/mL) during 24 h, the viability of BHK-21, FEA and VERO cell lines were evaluated by MTT assay. Viability of BHK-21 cells significantly (P < 0.001) decreased after treatment with 3500; 5500 and 7500 µg/mL of dihydrostreptomycin and 9000; 10000 and 20000 µg/mL of neomycin. The FEA cell viability decreased significantly (P < 0.001; P < 0.01) at 2500 and 3000 µg/mL dihydrostreptomycin and at 3000 µg/mL of neomycin treatment. Only the highest concentration of dihydrostreptomycin (20000 µg/mL) reduced VERO cell viability significantly (P < 0.01). Based on or results we can assume the effect of different antibiotics in different concentrations on cell lines is various. Detection of antibiotic toxicity to animal cells is very important because of the increasing resistance of bacteria. One of the solutions is drug dose increasing, but only to a certain concentration, since the toxic effect over the therapeutic one will prevail, which we have also shown in this work.

Events occurring during the previous lactation, the dry period, and peripartum as risk factors for early lactation mastitis in cows receiving 2 different intramammary dry cow therapies.

The objective of this study was to investigate the association between mastitis events occurring during the previous lactation, the dry period, and the peripartum period on the incidence of early lactation mastitis in cows receiving ceftiofur hydrochloride or penicillin dihydrostreptomycin as intramammary dry cow antibiotic therapy. Cows (n=402) from 2 large dairy farms in Central Florida were enrolled in the study at the time of dry-off processing and were randomly assigned to 1 of 2 dry cow therapies: ceftiofur hydrochloride or penicillin dihydrostreptomycin. Composite milk samples were collected at dry-off and after calving for bacteriological examination and somatic cell count. Peripartal health disorders were monitored during the first 30 d of lactation and included calving difficulty, metritis, ketosis, and left displaced abomasum. Milk production and individual somatic cell scores (SCS) were recorded monthly by the Dairy Herd Improvement Association. The main outcome variables were the risk of clinical mastitis during the first 30 and 60 d of lactation, and the risk of subclinical mastitis at the first 2 monthly Dairy Herd Improvement Association tests after calving (up to 70 d in milk). Additionally, the SCS and the presence of mastitis pathogens in milk at dry-off and at calving were analyzed. Explanatory variables consisted of events occurring during the previous lactation, at dry-off and during the dry period, at calving, and within the first 30 d after calving. Multiple events occurring during the previous lactation had a significant effect on the incidence of mastitis in the subsequent lactation. These events included low milk yield, intermediate lactation length, clinical mastitis, and lactation SCS average. Similarly, intramammary infections with environmental bacteria at dry-off increased the chances of clinical mastitis the first month after calving. Dry-off therapy had a significant effect on mastitis incidence; cows treated with ceftiofur hydrochloride had lower odds of having clinical and subclinical mastitis in the subsequent early lactation compared with cows treated with penicillin dihydrostreptomycin.

Streptococcus dysgalactiae isolates at calving and lactation performance within the same lactation.

The objective of the study was to investigate the association between early lactation Streptococcus dysgalactiae isolates and milk yield, somatic cell count (SCC), clinical mastitis, and culling in the same lactation. The 178 commercial dairy herds were randomly placed into 3 penicillin- or penicillin-dihydrostreptomycin-based dry-cow treatments and 3 different postmilking teat disinfection groups-negative control, iodine, or external teat sealant. All cows were sampled in early lactation, and Strep. dysgalactiae-positive and culture-negative cows were followed throughout the remainder of the lactation. Mixed models, including repeated measurements, with test-day observation as dependent variable, were used to compare milk yield, SCC, and available milk quality variables throughout the remaining lactation. Survival analyses, using a positive frailty model to account for any herd random effects, were used to estimate the hazard ratio for clinical mastitis and culling. Streptococcus dysgalactiae-positive cows had a significantly higher SCC throughout the lactation compared to culture-negative cows. For primiparous or multiparous cows, respectively, the differences in the geometric mean SCC between Strep. dysgalactiae-positive and culture-negative cows was 197,000 or 280,000 cells/mL at the beginning of the lactation, 24,000 or 46,000 cells/mL in mid lactation, and 39,000 or 111,000 cells/mL at the end of the lactation. Streptococcus dysgalactiae-positive primiparous or multiparous cows produced 334 or 246 kg less milk, respectively, during a 305-d lactation compared with culture-negative cows. Compared with culture-negative cows, the hazard ratios for clinical mastitis in Strep. dysgalactiae-positive cows were 2.3 (1.9 to 2.9) and 1.6 (1.3 to 2.0) for culling. For cows with both Strep. dysgalactiae and Staphylococcus aureus isolates, the hazard ratio for culling significantly increased to 2.5 (1.9 to 3.2).

Clinical mastitis in Norwegian herds after a combined selective dry-cow therapy and teat-dipping trial.

The objective of this study was to see if introduction of a 2-yr combined selective dry-cow therapy and teat-dipping trial would reduce clinical mastitis (CM) events in 164 Norwegian dairy herds. Three different penicillin or penicillin/dihydrostreptomycin-based dry-cow treatments, and 3 different teat-dipping regimens (negative control, iodine teat dip, or an external teat sealant) were independently and randomly allocated to each herd. Complete lactations both before and during the trial were investigated. Altogether, 1,005 CM cases were recorded in the lactations before the trial and 924 cases were recorded during the trial. Bacteriological milk samples were available from 784 of the 924 CM cases during the trial. Among these, Staphylococcus aureus were isolated from 47.4%, Streptococcus dysgalactiae from 22.5%, Escherichia coli from 10.7%, and coagulase-negative staphylococci from 6.3%. In addition, 12.5% cases were bacteriological negative, and the remainder of the CM cases were caused by other microbes. The different models were analyzed using Cox regression analysis with PROC PHREG and a positive stable frailty model in the SAS macro. Separate models were made for cows housed in tie-stalls and free-stalls. Parity had a significant impact on the CM risk in both type of stalls. Older cows (parity > 3) had the highest hazard ratio of contracting CM in tie-stalls (1.68) and free-stalls (2.18) compared with parity 1. The CM risk decreased significantly (13%) in tie-stalls and by 18% in free-stalls. In tie-stalls, iodine-dipped cows had a significantly lower chance (21%) of getting CM compared with the negative control and the use of external teat sealant. The same trend was seen in free-stalls; however, the differences were not significant. Compared with CM before the trial, the reduction of CM was 15% during the trial.

The comparative pharmacokinetics of 3H-dihydrostreptomycin in solution and liposomal form in normal and Mycobacterium tuberculosis infected mice.

In intact and tuberculous mice a comparative pharmacokinetics of 3H-dihydrostreptomycin (DHS) encapsulated into the liposomes (LE) prepared from egg lecithin and soluble form of DHS (SOL) after their intravenous administration has been studied. The values of the total area under the serum concentration--time curve of DHS (AUC 0----infinity), administered in the LE form was 8.8 times higher in normal and 5.9 times higher in mice with advanced tuberculosis than in mice treated with soluble form of DHS (SOL). AUC 0----infinity values in spleen and liver were 2.2 and 4.5 times higher, respectively, in use of LE form, where as AUC 0----infinity values in lungs and kidneys were independent from the drug form of DHS in normal mice. DHS levels in lungs of tuberculous mice were 3 times higher 3 hours after administration of LE in comparison with those after SOL DHS. AUC 0----infinity values in spleen and liver of tuberculous mice were 9.2 and 7.3 times higher, respectively, in comparison with those after SOL DHS. AUC 0----infinity values in kidneys of tuberculous mice didn't differ significantly independently from the use of one or another form of the drug.

Antibiotics and steroids in the treatment of acquired subglottic stenosis. A canine model study.

The efficacy of a combination of systemic antibiotics and a steroid in the prevention of acquired subglottic stenosis (ASGS) was evaluated employing a previously developed canine animal model. Thirty-five healthy, postweanling mongrel puppies aged 5 weeks were each intubated for 14 days with an uncuffed polyvinyl endotracheal tube. Twenty puppies received intramuscular dexamethasone (1 mg) daily, and procaine penicillin (100,000 IU) and dihydrostreptomycin (0.125 g) in two divided doses on the day of intubation and each day thereafter until the completion of the study. The remaining 15 puppies served as a control group and received no medical therapy. Animals from both groups were killed at 5, 7, 12, 15, 20, 30, and 56 days following intubation. Comparative examinations of the laryngotracheal complexes of treated and control dogs showed that there was no significant difference between the two groups in the ultimate degree of ASGS attained, as determined by intraluminal cross-sectional area analysis, or in the extent of the lesion as documented by gross and microscopic histology. This investigation showed that the specific systemic combination of two antibiotics and a steroid used in the study was not efficacious in the prevention of ASGS in a canine animal model; for this reason we question the benefit of analogous medical regimens employed to prevent ASGS in infants and children who require long-term endotracheal intubation.

The patas monkey as a model for dihydrostreptomycin ototoxicity.

Although the cochlear toxicity of dihydrostreptomycin (DHSM) is well-recognized in man, it has always proved difficult to demonstrate in animals. Hearing thresholds in M. nemestrina monkeys remained essentially unchanged after DHSM 100 mg/kg im daily for 8 months, but E. patas monkeys were severely deafened by DHSM 20 mg/kg for 90 days, a regimen formerly used in treating human tuberculosis. The patas monkey may prove to be the animal model of choice for evaluating aminoglycoside ototoxicity.

Effectiveness of various therapeutic regimens for bovine brucellosis.

Various chemotherapeutic regimens were evaluated in 48 culture-positive dairy cows. Cessation of shedding of Brucella abortus from udder secretions and absence in selected tissues at necropsy were criteria of success. A combination of a long-acting oxytetracycline and streptomycin eliminated Brucella in 10 of 14 (71.4%) cows. Two cows that were retreated with the same regimen also became culture-negative. Other treatment regimens, including the use of liposome-encapsulated antibiotics, were less successful. Serotests were a poor criterion of effectiveness.

Effect of combined antibiotic therapy on fertility in brood bitches infected with Brucella canis.

Bitches with naturally occurring Brucella canis infection were treated with combined antibiotic therapy consisting of tetracycline, dihydrostreptomycin, and trimethoprim-sulfadiazine. After treatment, all but 1 bitch became abacteremic, and serologic titers declined for a variable length of time (3 months to 1 years). Abortion did not occur while these bitches were abacteremic. Although sequential antibiotic therapy for 6 weeks did not eradicate Brucella canis from affected bitches, it did not prevent abortion. The number of live pups whelped and weaned by treated bitches was comparable with that in bitches before they became infected.

Comparative efficacy of three antibiotic products for the treatment and prevention of subclinical mastitis during the dry period.

The comparative efficacy of a product containing 500 mg benzathine cloxacillin (Orbenin D.C.). another product containing 100 mg nafcillin, 300 mg procaine benzylpenicillin and 100 mg dihydrostreptomycin (Nafpenzal D.C.) and a third product containing 250 mg cephalonium (Cepravin D.C.), in the elimination and prevention of mastitis due to gram-positive microorganisms during the dry period was investigated in 1253 cows located in 14 herds in Israël. Although more uniform results were observed after treatment with Nafpenzal D.C. differences among herds were rather large. The mean cure rate for Staphylococcus aureus quarter infections was 81.4 per cent. New S. aureus infections which occurred between drying off and post calving sampling time were found in 7.0 per cent of the quarters. Dry period therapy reduced infection level with S. aureus from 13.5 per cent of quarters to 8.0 per cent. A similar proportional decrease was found in infection level due to non-agalactiae Streptococcus quarter infections.

Clinical trial of three therapeutic regimens for bovine mastitis.

An open, block randomised multi-centre clinical trial was performed in Norway during 1985 to 1987 to compare the therapeutic efficacy of three antibiotic regimens against clinical bovine mastitis caused by penicillin-sensitive bacteria. Two regimens consisted of procaine penicillin injected intramuscularly for either three or five days, and the third, the traditional Norwegian regimen, consisted of one intramuscular injection of a combination of procaine penicillin and dihydrostreptomycin followed by one intramammary treatment daily per infected quarter for four days. The study included 621 quarters with infectious mastitis from 439 cows. The most efficient regimen for all bacteria was five days systemic treatment (53.1 per cent cured), and the traditional regimen was second best (46.7 per cent cured). The least efficient regimen consisted of systemic therapy with procaine penicillin for three days (36.9 per cent cured). The difference between the therapeutic efficacies of the three regimens was reduced when the clinical mastitis was severe, and in severe mastitis caused by Staphylococcus aureus the difference was very small.

Antibiotic levels in bovine lacrimal fluid after single application of ointments containing procaine benzyl penicillin plus dihydrostreptomycin; and benzathine cloxacillin.

The efficacy of an experimental slow-release formulation, containing procaine benzyl penicillin and dihydrostreptomycin, was investigated in a cross-over study with a cloxacillin eye ointment in 12 cows with clinically normal eyes. After a single topical application the therapeutic concentrations of penicillin were sustained for 48 to 92 hours and of cloxacillin for 32 to 48 hours. These long-acting ointments will simplify the successful treatment of painful eye disorders such as keratoconjunctivitis. A practical and non-irritant method for sampling tears is described.

Comparison of two regimens for the treatment of clinical bovine mastitis caused by bacteria sensitive to penicillin.

The efficacies of two regimens for the treatment of acute clinical mastitis were compared in a randomised multi-centre field trial in Norway, using 657 cows. The purpose was to determine whether repeated intramuscular injections of penicillin G for three days were more effective than a single injection, when given in combination with intramammary treatment for five days. The results were evaluated on the basis of clinical and microbiological examinations and cell count determinations of quarter milk samples taken at the initial visit and four weeks later. There were no significant differences between the effects of the treatments, either for all the cows, or for subgroups of the cows based on age, stage of lactation, and systemic reaction, or the type of causal bacteria.

Treatment of clinical mastitis: two intramammary formulations compared.

Two hundred cases of mild clinical mastitis were treated on two farms using two intramammary preparations in quick release bases. One preparation contained penicillin and streptomycin, the other contained lincomycin, neomycin and prednisolone. Fifty-eight per cent of cases were clinically and bacteriologically cured using the first preparation; and 61 per cent of cases, with a similar range of organisms, using the second. The main infections were Streptococcus uberis and coliform; all but three of the infections were sensitive in vitro to one or both of the antibiotics in the preparations. On the farm with 100 cows, 69 per cent of cases were cured, while only 52 per cent were cured on the farm with 300 cows. It is suggested that it is more difficult to detect and treat cases in the larger herd. A number of reservations about the interpretation of the results are discussed.

Presence of dihydrostreptomycin and penicillin in cows' milk following intrauterine administration.

Cows were given liquid intrauterine infusions of dihydrostreptomycin and penicillin or dihydrostreptomycin alone approximately 24 hours after the onset of oestrus to determine the necessary withholding period for milk intended for human consumption. When dihydrostreptomycin and procaine penicillin were administered together, a withholding period of 72 hours provided an adequate safety margin. Thirty-six hours was an adequate withholding period when dihydrostreptomycin was administered.

Successful treatment of mycoplasmosis in layer chickens with single dose therapy.

The efficacy of treatment with single dose administration of 5 drugs at different dosages to layer hens naturally infected with Mycoplasma gallisepticum was studied. The drugs were tiamulin, which was administered orally, tylosin (parenterally and orally), spiramycin (orally), long-acting oxytetracycline (parenterally) and tylosindihydrostreptomycin (parenterally). Cure was assessed by the absence of nasal discharge. The cure rate was significantly higher (P less than 0.05) in treated hens than in untreated hens, as early as 1 day after treatment. Remission for 33 days was achieved in 60% of hens treated with 100 mg oxytetracycline, in 100% of hens treated with 100 mg or 200 mg spiramycin, in 92% and 85% of hens treated with 100 mg tylosin, parenterally and orally, and in 89% and 88% of birds given 100 mg tiamulin and tylosin-dihydrostreptomycin, respectively.

Amoxycillin as an alternative to dihydrostreptomycin sulphate for treating cattle infected with Leptospira borgpetersenii serovar hardjo.

OBJECTIVE: To assess the effect of amoxycillin treatment on urinary excretion of leptospires from cattle infected with Leptospira borgpetersenii serovar hardjo. DESIGN: A chemotherapy trial with controls. PROCEDURE: Fourteen heifers serologically negative to L hardjo were inoculated with L hardjo via the conjunctival route and assessed for evidence of infection by serological, fluorescent antibody and microbiological tests. Two injections (48 h apart) of amoxycillin at a dose of 15 mg/kg were administered intramuscularly to seven heifers 6.5 weeks after infection; the remaining heifers acted as untreated controls. Later, these seven control group heifers were treated with a single dose of amoxycillin (15 mg/kg). Samples of urine were collected before and after amoxycillin treatments; kidneys were collected at slaughter, and examined by fluorescent antibody test and microbiological culture. RESULTS: Leptospires were isolated from the urine of 11 of 14 heifers inoculated with L hardjo. After treatment of six of these with two injections of amoxycillin, leptospires were not isolated. Of the controls, four of the five initially leptospiruric heifers continued to shed leptospires; after a single injection of amoxycillin, no leptospires were detected in the kidneys of these four. CONCLUSION: Amoxycillin may be an acceptable alternative to dihydrostreptomycin sulphate for the treatment of cattle infected with L hardjo.

An unusual presentation of suspected oedema disease of swine in Kenya.

From a group of 11 recently weaned pigs, 4 were reported to be sick. Clinical examination of the sick pigs revealed marked dyspnoea, bluish-red discolouration of the skin, incoordination and difficulty in walking. Bacteriological examination of the gut contents of 2 pigs that had died earlier yielded pure cultures of haemolytic Escherichia coli. Post mortem examination of the remaining 2 pigs that died subsequently revealed progressive pulmonary collapse. One of these also showed subcutaneous oedema of the head and marked oedema of the mesentery of the spiral colon and oedema of the brain. Microscopically there was pulmonary alveolar collapse and degenerative changes in the liver. On the basis of the clinical signs, isolation of haemolytic E. coli and the post mortem findings, a diagnosis of oedema disease was made.