In vitro Elution of Amikacin and Vancomycin from Impregnated Plaster of Paris Beads

Objective: To describe in vitro elution characteristics of amikacin and vancomycin from calcium sulfate hemihydrate 98% (plaster of Paris, POP) beads and characterize eluent inhibition of Staphylococcus spp. Study Design: Experimental study. Methods: POP beads were impregnated with amikacin or vancomycin alone or in combination and then incubated alone or in combination for 84 days at 37°C in plastic tubes containing sterile phosphate‐buffered saline (PBS). Beads containing no antimicrobial served as negative control. Beads were intermittently moved to a new tube containing drug‐free PBS. Antimicrobial was measured in the eluent using a polarized fluorescent immunoassay. Eluent inhibition of Staphylococcus spp. was determined at each time point. Results: Antimicrobial release from beads was characterized by an initial rapid phase then a slower phase. Although antimicrobial release from beads occurred throughout the 84 days, most was in the first 24 hours, except for vancomycin alone. Duration of eluent inhibition of Staphylococcus spp. growth ranged from 0.5 (amikacin alone) to 56 days (vancomycin alone). Control eluent did not inhibit bacterial growth. Conclusions: Amikacin elution from POP beads was rapid, inhibiting growth for <24 hours with or without vancomycin. Vancomycin elution was slower and inhibited growth for 56 days alone or for 5 days with amikacin. Clinical Relevance: Vancomycin‐impregnated beads appear to be reasonable as a therapeutic option whereas amikacin‐impregnated POP beads and amikacin and vancomycin combinations may require further study before considering as a therapeutic option.     

Elution of antimicrobials from a cross‐linked dextran gel: In vivo Quantification

Reasons for performing study: Use of a novel, biodegradable, antimicrobial‐impregnated gel provides an alternative method of local treatment of infections in horses. Objectives: To determine in vivo elution of antimicrobial medications from antimicrobial‐impregnated cross‐linked dextran gel and to evaluate the effect on wound healing when implanted subcutaneously in horses. Methods: Amikacin‐, vancomycin‐ or amikacin/clindamycin‐impregnated gel was placed subcutaneously in 11 horses' necks, using 6 replicates with a 3 month washout between experiments. Capillary ultrafiltration probes for collection of interstitial fluid were placed 0 cm and 1.5 cm from the gel‐filled incisions. Samples were collected at 0, 4, 8 and 12 h, and on Days 1–10. Blood was collected on Days 0, 1 and 7. Amikacin and vancomycin samples were analysed via fluorescence polarisation immunoassay, and clindamycin samples via high‐performance liquid chromatography. Histology of biopsy samples was performed at the completion of the study. Differences in mean histomorphological scores between groups were assessed using Wilcoxon's signed ranks test. Results: Maximum antimicrobial concentrations were detected at 4 h (amikacin), and 8 h (vancomycin, and amikacin and clindamycin from the combination gel). Mean ± s.d. peak concentrations for amikacin, vancomycin, amikacin (amikacin/clindamycin) and clindamycin were 6133 ± 1461, 7286 ± 2769, 3948 ± 317 and 985 ± 960, respectively. Median number of days for which antimicrobial concentration remained above minimum inhibitory concentration for target microorganisms at implantation was ≥10 days for vancomycin, 9 days (± 1) for amikacin and 8 days (± 1) for clindamycin. Mean plasma amikacin and vancomycin concentrations were lower than detectable limits; mean serum clindamycin concentrations were 0.52 µg/ml and 0.63 µg/ml at 24 h and 7 days, respectively. There were no significant differences in histomorphological scores between treatment and control incisions (P≥0.22). Conclusions and potential relevance: Cross‐linked dextran gel is a safe, effective alternative local antimicrobial delivery method.     

In vitro elution of amikacin and ticarcillin from a resorbable, self-setting, fiber reinforced calcium phosphate cement

Objective: To determine in vitro elution characteristics of amikacin and ticarcillin from fiber reinforced calcium phosphate beads (FRCP). Sample Population: Experimental. Methods: FRCP beads with water (A), amikacin (B), ticarcillin/clavulanate (C), or both amikacin and ticarcillin/clavulanate (D) were bathed in mL phosphate‐buffered saline (PBS) at 37°C, 5% CO₂ and 95% room air. PBS was sampled (eluent) and beads were placed in fresh PBS at time points 1 and 8 hours and 1, 2, 3, 4, 5, 6, 7, 10, 12, 14, 18, 21, 25, 28, 35, 42, 49, and 56 days. Antibiotic concentration and antimicrobial activity of eluent against Escherichia coli, Staphylococcus aureus, and Klebsiella pneumoniae were determined. Results: Both antibiotics eluted in a bimodal pattern. Beads with a single antibiotic eluted 20.8 ± 2.5% of amikacin and 29.5 ± 0.8% of ticarcillin over 56 days. Coelution of the antibiotics resulted in a lower proportion (AUC_0–∞) of antibiotics eluted for both amikacin (9.5 ± 0.2%) and ticarcillin (21.7 ± 0.09%). Bioassay of antimicrobial activity of the eluent (t=1, 8, and 24 hours) established reduced antimicrobial activity of amikacin from combination beads (D). Conclusions: FRCP beads with amikacin or ticarcillin/clavulanate, but not the combination, are suitable carriers for wound implantation. Clinical Relevance: Duration before complete resorption of FRCP beads in vivo should be determined before clinical use as a resorbable depot. The results of this study underscore the importance of testing drug combinations, despite success of the combination systemically, before their use in local applications.     

In Vitro and In Vivo Evaluation of Ferric-Hyaluronate Implants for Delivery of Amikacin Sulfate to the Tarsocrural Joint of Horses

Objective— To assess the antimicrobial elution characteristics, toxicity, and antimicrobial activity of amikacin‐impregnated ferric‐hyaluronate implants (AI‐FeHAI) for amikacin delivery to the tarsocrural joint of horses. Study Design— Experimental study. Sample Population— AI‐FeHAI implants, equine cartilage, and synovium, and horses (n=6). Methods— In vitro study: Five AI‐FeHAI were placed in saline solution with daily replacement until implant degradation. Eluent was tested for amikacin concentration and bioactivity. Synovial and cartilage explants were incubated in the presence or absence of AI‐FeHAI for 72 hours and subsequently assessed for morphology, viability, and composition. Synovial explants were incubated with Staphylococcus aureus in the presence or absence of AI‐FeHAI. Spent medium was cultured daily and explants were assessed for morphology and viability after 96 hours. In vivo study: AI‐FeHAI were placed in 6 tarsocrural joints. Standard cytologic analysis and amikacin concentration (SFAC) were determined in synovia obtained regularly for 28 days thereafter. Similar analyses were conducted after a single intra‐articular injection of amikacin 6 months later. Results— In vitro study: Amikacin concentrations exceeded 16 μg/mL and inhibited S. aureus growth for 8 days. AI‐FeHAI had no effect on cartilage explants. AI‐FeHAI eliminated bacteria from synovial explants. In vitro study: After AI‐FeHAI placement, SFAC was highest (140.78+63.81 μg/mL) at first sampling time. By 24 hours SFAC was <16 μg/mL. After intra‐articular injection, SFAC was the highest (377.91 ± 40.15 μg/mL) at first sampling time. By 48 hours SFAC was <16 μg/mL. Conclusions— A single intra‐articular amikacin injection demonstrated superior pharmacokinetics than AI‐FeHAI prepared as described. Clinical Relevance— AI‐FeHAI cannot be recommended for clinical use.     

In vitro elution studies of amikacin and cefazolin from polymethylmethacrylate

OBJECTIVE: To compare the in vitro elution characteristics of amikacin and cefazolin from polymethylmethacrylate (PMMA) alone and in combination. STUDY DESIGN: A prospective, controlled, experimental study. METHODS: Three aliquots of 6 g sterile PMMA were measured and to them added (1) 750 mg amikacin; (2) 1050 mg cefazolin; and (3) 750 mg amikacin and 1050 mg cefazolin. Ten beads of each antimicrobial/PMMA combination were placed in 5 mL phosphate-buffered saline (PBS) at pH 7.4 and room temperature with constant agitation. PBS was sampled at 15 time points between 1 hour and 30 days. Amikacin concentrations were determined by fluorescence polarization immunoassay and cefazolin concentrations by high-performance liquid chromatography. RESULTS: Amikacin and cefazolin eluted at concentrations greater than 8 and 4 times, respectively, above the minimum inhibitory concentration (MIC) for susceptible bacteria over 30 days. Co-elution of the antibiotics resulted in a greater rate and proportion of antibiotic eluted. Concentrations of amikacin and cefazolin in the co-eluted fluid were not maintained sufficiently above the MIC for selected bacteria over 30 days. CONCLUSIONS: PMMA beads of only amikacin or cefazolin-eluted concentrations greater than the MIC for selected bacteria for 30 days. Co-elution of the antibiotics at the selected doses resulted in a significantly shorter duration of elution and may not be effective for treatment of wound infection. CLINICAL RELEVANCE: Co-elution of amikacin and cefazolin from PMMA at the selected doses cannot be recommended for sustained treatment of infection.     

Methicillin‐resistant Staphylococcus aureus keratitis in a dog

The purpose of this study is to report a case of methicillin‐resistant Staphylococcus aureus (MRSA) keratitis in a dog. A 7‐year‐old intact male American cocker spaniel that had undergone removal of a nictitating gland was referred for severe ulcerative keratitis. Slit‐lamp examination showed swelling of the eyelid, mucopurulent discharge, conjunctival injection and chemosis, diffuse corneal edema and opacity, and a deep ulcer in central cornea. Gram staining of discharge from the eye demonstrated Gram‐positive cocci. Despite topical ofloxacin, oxytetracycline and polymyxin B ophthalmic solution and intravenous cefazolin, there was no improvement. Cultures revealed MRSA that was sensitive only to chloramphenicol, vancomycin, lincomycin, and clindamycin. The antibiotic regimen was changed to topical and systemic chloramphenicol. After 9 days of treatment, although inflammation started to be resolved, the dog developed nonregenerative anemia. The antimicrobial regimen was changed again to topical and systemic vancomycin. Inflammation continued to improve over the next week. MRSA should be considered a potential organism in infectious keratitis, especially when general antibiotics are not effective. Although topical and systemic chloramphenicol and/or vancomycin are effective for treating MRSA keratitis, vancomycin should only be used when culture and susceptibility results indicate it is appropriate and no other options are available. To our knowledge, this is the first detailed case report of MRSA keratitis in a dog.     

In vitro elution of gentamicin, amikacin, and ceftiofur from polymethylmethacrylate and hydroxyapatite cement

OBJECTIVE: To compare the elution characteristics of ceftiofur and liquid and powdered gentamicin and amikacin from polymethylmethacrylate (PMMA) and from hydroxyapatite cement (HAC). METHODS: PMMA and HAC beads in triplicate were impregnated with various amounts and formulations of antibiotics. Beads were immersed in 5 mL of phosphate buffered saline that was replaced at 1, 3, 6, and 12 hours, and 1, 2, 3, 5, 7, 10, 14, 18, 22, 26, and 30 days. The eluent was stored at -70 degrees C until assayed within 2 weeks by microbiological assay (gentamicin and amikacin) or capillary electrophoresis (ceftiofur). RESULTS: Rate of elution for all beads was greatest within the first 24 hours. Cumulative release of total antibiotic dose from beads over 30 days was significantly greater from HAC than PMMA. Antibiotic elution was directly related to the amount of antibiotic incorporated into the cement. Powdered and liquid forms of gentamicin had similar elution rates from PMMA. Elution of amikacin from PMMA beads was greater when the powdered form was used compared with liquid amikacin. Eluent concentrations of ceftiofur were similar to those of the aminoglycosides during the first 3 to 7 days but then decreased precipitously by comparison. CONCLUSIONS: Elution of antibiotics from HAC was greater than from PMMA. Gentamicin- and amikacin-impregnated PMMA and HAC released bactericidal concentrations of antibiotic for at least 30 days. Ceftiofur-impregnated PMMA or HAC is unlikely to provide long-term bactericidal concentrations. CLINICAL RELEVANCE: Gentamicin and amikacin elute effectively from PMMA and HAC.     

Effect of estradiol‐17β during in vitro growth culture on the growth,maturation, cumulus expansion and development of porcine oocytes from early antral follicles

Growing porcine oocytes from early antral follicles can acquire meiotic and developmental competence under suitable culture conditions, but at lower rates compared to full‐grown oocytes. We postulated that estradiol‐17β (E₂) supported the acquisition of meiotic and developmental competence as well as cumulus‐expansion ability during growth culture. Growing oocytes from early antral follicles (1.2 to 1.5 mm in diameter) were grown in vitro for 5 days in a medium containing 0, 10^?7, 10^?6, 10^?5 or 10^?4 mol/L E₂; after in vitro maturation, 35, 58, 47, 74 and 49% of oocytes matured to metaphase II, 25, 79, 77, 90 and 97% acquired cumulus‐expansion ability, and 23, 54, 63, 89 and 64% were fully surrounded by cumulus cells, respectively. Following maturation, electro‐stimulation was applied to the oocytes grown with 10^?5 mol/L E₂. After 6 days of culture, in vitro‐grown oocytes developed to the blastocyst stage at a rate similar to that for full‐grown oocytes (31% and 40%, respectively). Therefore, we suggest that the use of E₂ during growth culture improves the meiotic and developmental competence of oocytes, cumulus‐expansion ability, and cumulus cell attachment to the oocytes.     

Microbiological Features of Staphylococcus schleiferi subsp. coagulans,Isolated from Dogs and Possible Misidentification with Other Canine Coagulase‐positive Staphylococci

Staphylococcus schleiferi subsp. coagulans has only rarely been isolated and identified from the external auditory meatus of dogs suffering from external otitis. Its morphological and basic biochemical characteristics are of relatively little value for identification, as it phenotypically resembles another coagulase‐positive staphylococci (CPS) and, consequently, may be easily misidentified as S. intermedius or even as S. aureus. In the present work, differentiation of S. schleiferi ssp. coagulans was therefore based on specific biochemical and genetic methods. All the strains were evaluated with the following commercial methods: Api Staph System (bioMérieux, Marcy l'Etoil, France), BBL Crystal Identification Systems (Gram‐Positive ID Kit and Rapid Gram‐Positive ID Kit; Becton Dickinson), and GEN‐PROBE^® AccuProbe, Staphylococcus aureus identification test (bioMérieux). Gram‐Positive ID System/GP database includes the broadest range of staphylococcal species and correctly identifies the majority of strains important in veterinary medicine. Therefore, it is an acceptable alternative to conventional methods for identification of canine staphylococcal isolates. Reliable differentiation of S. aureus from S. schleiferi ssp. coagulans and S. intermedius was feasible with AccuProbe for S. aureus, which gave positive results only for S. aureus; all other CPS tested were negative.     

The effect of three tropical medicinal plants and superzist probiotic on growth performance,carcass characteristics,blood constitutes,immune response,and gut microflora of broiler

The pivotal aim of the present context was to determine the effect of probiotic (superzist) and medicinal plants (R. coriaria, H. persicum, and M. piperita) powder on growth performance, carcass characteristics, blood parameters, immunity response, and microflora counts of broilers over a 42-day feeding period. One hundred and fifty one-day-old chickens were randomly allocated to the following five treatments: (1) Control diets; (2) control diets?+?0.03% w/v superzist; (3) control diets?+?0.5% w/v R. coriaria; (4) control diets?+?0.5% w/v H. persicum; and (5) control diets?+?0.5% w/v M. piperita. No significant differences among treatments on broiler performance during 29th to 35th days of age, whereas a gradual reduction (P?<?0.05) in the feed efficiency and economic index values between 36th to 42nd days of age, across the treatments versus control diet. Supplementations were increased (P?<?0.05), the colon weight, colon length, and right cecum weight versus control. However, the superzist and phytogenic supplementation slightly increased the jejunum length, ileum length, and left cecum length. The supplementation showed reduced heterophils, increased lymphocytes and monocytes percentage, and decreased heterophils/lymphocytes ratio. Additions of superzist and medicinal plants reduced (P?<?0.05) the total counts of E. coli and improved (P?<?0.05) the cecal lactobacilli. In conclusion, under the appropriate conditions of this investigation, the superzist and tested medicinal plants could undoubtedly be ideal alternatives to the use of existing antibiotics as growth promoters in poultry industry.

     

Effect of Antibiotics in Extender on Bacterial and Spermatozoal Quality of Cooled Buffalo (Bubalus bubalis) Bull Semen

The present study was designed to study the effect of traditional antibiotic combination (streptomycin and penicillin; SP) and relatively modern combination of antibiotics (gentamycin, tylosin, lincomycin and spectinomycin; GTLS) in extender on bacterial control and spermatozoal quality of liquid buffalo bull semen stored at 5°C. Semen collected from Nili‐Ravi buffalo bulls (n = 10) was diluted with skim milk extender containing either SP (streptomycin 1000 μg/ml and penicillin 1000 IU/ml), GTLS (gentamycin 500 μg/ml, tylosin 100 μg/ml, lincomycin 300 μg/ml and spectinomycin 600 μg/ml) or negative control with no antibiotics (NA). Liquid semen was stored at 5°C for 5 days. Aerobic bacteria isolated from buffalo semen were Pseudomonas aeruginosa and Staphylococcus aureus. The only facultative anaerobic bacterium isolated was Klebsiella pneumoniae. In vitro antibiotic sensitivity test revealed that Ps. aeruginosa and Staph. aureus were susceptible to gentamycin. Staphylococcus aureus and K. pneumoniae were susceptible to tylosin and linco‐spectinomycin. Total aerobic bacterial count was significantly lower in semen samples treated with GTLS than those of SP on third and fifth day of storage at 5°C. There was no difference (p > 0.05) in sperm motility, longevity and plasma membrane integrity (PMI) in extender containing SP or GTLS combination until the third day of storage at 5°C. On fifth day of storage sperm motility, longevity and PMI was significantly better in extender containing SP compared with GTLS and NA. Intact acrosomes, and sperm head, mid piece and tail abnormalities remained similar (p > 0.05) because of antibiotics up to 5 days of storage. In conclusion, GTLS is more capable than SP for bacterial control of buffalo bull semen. Moreover, GTLS and SP are equally efficient in preserving spermatozoal quality of extended buffalo bull semen for 3 days at 5°C.     

The survival of multidrug‐resistant bacteria in thermophilic and mesophilic anaerobic co‐digestion of dairy manure and waste milk

Anaerobic digestion is considered as a promising method to manage animal waste with antibiotic‐resistant bacteria. Current research was conducted to investigate the survival of multidrug‐resistant bacteria (MDRB) resistant to three groups of antibiotics: (i) cefazolin, neomycin, vancomycin, kanamycin (group 1); (ii) penicillin, oxytetracycline, ampicillin, streptomycin (group 2); and (iii) cefazolin, neomycin, vancomycin, kanamycin, penicillin, oxytetracycline, ampicillin, streptomycin (group 3), in anaerobic digestion of dairy manure and co‐digestion of dairy manure and waste milk at 37°C and 55°C for 22 days, respectively. The population densities of three groups of MDRB on peptone, tryptone, yeast and glucose agar plates incubated at 30°C for 7 days before and after digestion showed 100% destruction in both digestates at thermophilic temperature. Overall reduction of more than 90% of three groups of MDRB was observed in mesophilic digestion with no significant differences (P > 0.05) between manure and milk mixture. Co‐digestion of dairy manure and waste milk always produced significantly (P < 0.05) higher total gas and methane gas than digestion of manure alone at both temperatures. Gas production in each case was significantly (P < 0.05) higher in thermophilic digestion than in mesophilic digestion. The results demonstrate that thermophilic co‐digestion of dairy manure and waste milk offers more benefits in terms of the environment and economy.     

Effects of leptin on the release of luteinizing hormone, growth hormone and prolactin from cultured bovine anterior pituitary cells

The effects of leptin on the release of luteinizing hormone (LH), growth hormone (GH) and prolactin (PRL) were studied in cultured bovine anterior pituitary (AP) cells in vitro. The AP cells were obtained from fully‐fed Japanese Black steers and were incubated for 3 h with 10^?13 to 10^?7 mol/L of leptin after incubating in Dulbecco's modified Eagle's Medium for 3 days. Leptin significantly increased the concentration of LH in the culture medium by 45 and 44% at doses of 10^?8 and 10^?7 mol/L, respectively, compared with the controls (P < 0.05). Leptin significantly increased the concentration of GH in the culture medium by 14 and 12% at doses of 10^?8 and 10^?7 mol/L, respectively (P < 0.05). Leptin also significantly increased the concentration of PRL in the culture medium by 26% compared with the controls at a dose of 10^?7 mol/L (P < 0.05). These results show that leptin stimulates the release of LH, GH and PRL by acting directly on bovine AP cells from fully‐fed steers.     

A phase II study to evaluate the toxicity and efficacy of alternating CCNU and high‐dose vinblastine and prednisone (CVP) for treatment of dogs with high‐grade,metastatic or nonresectable mast cell tumours

Safety and efficacy of a protocol of alternating 1‐(2‐chloroethyl)‐3‐cyclohexyl‐1‐nitrosourea (CCNU; 70 mg m^?2) and vinblastine (3.5 mg m^?2), and prednisone (1–2 mg kg^?1; CVP) in dogs with mast cell tumours (MCT) were evaluated. A total of 17 dogs had nonresectable MCTs and 35 received CVP as adjunctive treatment to locoregional control of metastatic MCTs or grade III MCTs. Neutropenia with fever occurred in 8% of dogs after treatment with vinblastine and in 2% after treatment with CCNU. Persistent elevation of serum alanine transaminase, suggestive of hepatotoxicity, occurred in 9% of the dogs. Response rate in dogs with nonresectable MCTs was 65%; five achieved a complete response (median, 141 days) and six achieved a partial response (median, 66 days). Overall median progression‐free survival (PFS) time in dogs treated in the adjuvant setting was 489 days. Dogs with grade III MCTs had shorter PFS compared with dogs with metastatic grade II MCTs (190 days versus 954 days; P < 0.001). Phase III studies are needed to provide reliable information about the comparative efficacy of this protocol.     

Effect of Intramammary Injection of RbIL‐8 on Milk Levels of Somatic Cell Count,Chemiluminescence Activity and Shedding Patterns of Total Bacteria and S. aureus in Holstein Cows with Naturally Infected‐subclinical Mastitis

The effect of intramammary injection of recombinant bovine interleukin‐8 (rbIL‐8, 1 mg/10 ml of saline) on quarter milk levels of somatic cell count (SCC), chemiluminescence (CL) activity and counts of total bacteria and Staphylococcus aureus (S. aureus) was investigated, using 10 Holstein cows with an early stage or a late stage of subclinical mastitis naturally infected with S. aureus. In the late‐stage group, milk SCC and CL activity had significant rises with maximum levels at 6 h, following maintained high levels thereafter post‐cytokine injection. The counts in milk total bacteria and S. aureus were insignificantly decreased, being increased back on day 7 post‐cytokine injection. Thus, the cytokine was inefficient for the late‐stage subclinical mastitis. However, in the early‐stage group milk SCC and CL activity declined to under pre‐injection levels on day 7 after marked and significant rises at 6 h and day 1 post‐cytokine injection. The milk total bacterial count decreased significantly on days 0.25 and 2. Furthermore, the milk S. aureus count was decreased significantly on days 1, 2, 3 and 7 by the cytokine injection. These results suggest that the rbIL‐8 has a potential as a therapeutic agent of the subclinical mastitis of dairy cows, if the cytokine is applied at an initial stage of infection.     

Genotypes,Antibiotic Resistance Profiles and Microarray‐Based Characterization of Methicillin‐Resistant Staphylococcus aureus Strains Isolated from Livestock and Veterinarians in Switzerland

Using different typing methods (MLST, spa‐, SCCmec‐ and agr‐typing), PFGE and DNA microarray‐based chip analysis, we characterized 20 MRSA strains isolated from livestock and veterinarians. PFGE analysis after macrorestriction with EagI provided seven different band patterns, which could be grouped into four clusters. One cluster consisted of all MRSA ST398 strains isolated from pigs, calves, mastitis milk and two veterinarians. One strain of ST398 from a veterinarian and the two strains of ST1 and ST8 formed the three other clusters. Antimicrobial susceptibility testing showed that 15 of 20 strains were resistant to ampicillin, cefoxitin, clindamycin, erythromycin, oxacillin, penicillin and tetracycline. All strains were susceptible to rifampin and vancomycin, 19 were susceptible to ciprofloxacin and 18 were susceptible to sulphamethoxazole/trimethoprim. Genes encoding different enterotoxins, leukotoxins and haemolysins were found in certain strains.     

The effect of dietary supplementation with Aurantiochytrium limacinum on lactating dairy cows in terms of animal health,productivity and milk composition

The aim of this study was to investigate the effect of dietary supplementation with the docosahexaenoic acid (DHA )‐rich microalgae, Aurantiochytrium limacinum (AURA ), on a variety of health and productivity parameters in lactating cows. Twenty‐four cows were blocked by parity and number of days in milk and then randomly assigned to a control (CON ; n  = 12) group with no algal supplementation, or a treatment group (AURA ; n  = 12) provided with 100 g AURA  cow^?1 day^?1 or 16 g DHA  cow^?1 day^?1. A variety of health and productivity measurements were taken, and results indicated that supplementation had no negative effects on animal health in terms of somatic cell count, haematological and biochemical blood parameters, while body condition was marginally improved by algal supplementation. No differences were found for the various production parameters measured; however, a tendency towards increased milk production was observed for the AURA group during the final stage of the study (+4.5 kg cow^?1 day^?1, day 78–84). The fatty acid profile of milk was improved by supplementation, with significantly lower saturated fatty acids, significantly higher omega‐3 fatty acids and an improved omega‐3/omega‐6 ratio observed when compared to the control group. The amount of DHA in the milk of cows provided 105 g AURA  head^?1 day^?1 was 4.7 mg/100 g milk with a peak transfer efficiency from feed to milk at day 49 of 8.3%. These results indicate that supplementation with 105 g AURA  head^?1 day^?1 resulted in the successful enrichment of milk with DHA without negatively impacting the health or productivity of the animals.     

Elution of metronidazole and gentamicin from polymethylmethacrylate beads

OBJECTIVE: To characterize the elution and bioactivity of metronidazole and gentamicin sulfate polymerized, individually and in combination, with polymethylmethacrylate (PMMA). STUDY DESIGN: In vitro experimental study. METHODS: PMMA beads containing metronidazole (3 concentrations), gentamicin sulfate, or metronidazole and gentamicin sulfate were immersed in 5 mL of phosphate-buffered saline in triplicate. Eluent was replaced at specified time intervals for 1 or 21 days, and antibiotic concentrations were measured by high-performance liquid chromatography. Changes in antibiotic bioactivity attributable to polymerization or copolymerization of the antibiotics with PMMA, ethylene oxide sterilization, and storage of AIPMMA beads containing metronidazole were evaluated. RESULTS: Antibiotic elution patterns were similar for all groups. Day 1 elution for groups containing metronidazole or gentamicin individually represented a mean 63%-66% and 79%, respectively, of the 21-day total. Approximately 50% of the day 1 elution occurred during the first hour. The elution of metronidazole was dose dependent. The elution of metronidazole (day 3-21) and gentamicin (all days) was significantly greater when metronidazole and gentamicin were combined (P <.05). The addition of metronidazole delayed polymerization of PMMA. Neither polymerization nor copolymerization of metronidazole and gentamicin with PMMA, gas sterilization, or 2-month storage of beads containing metronidazole significantly affected antimicrobial bioactivity. CONCLUSIONS: Metronidazole elution from PMMA was dose dependent. Copolymerization of metronidazole and gentamicin sulfate in PMMA resulted in increased rates of elution. Intraoperative preparation of metronidazole-impregnated PMMA beads is not practical, but sterilization and storage for 2 months should not affect efficacy. CLINICAL RELEVANCE: The local delivery of biologically active metronidazole and gentamicin by elution from PMMA is feasible.