Chlorhexidine Diacetate and Povidone-iodine Cytotoxicity to Canine Embryonic Fibroblasts and Staphylococcus aureus

Chlorhexidine diacetate and povidone-iodine were evaluated for fibroblast toxicity on a primary line of canine embryonic fibroblasts, and for bactericidal efficacy against Staphylococcus aureus. The cultured fibroblasts or S. aureus were exposed for 30 minutes to incremental dilutions of 0.5 and 0.0005% chlorhexidine diacetate, 5.0 to 0.05% povidone-iodine, or physiologic buffered saline as a control. To determine survival, fibroblasts were trypsinized and counted; S. aureus colonies were counted on brain-heart infusion agar. Survival for both groups was expressed by calculating the number of living cells in test dilutions as a percentage of the number in control cultures. Fibroblast survival occurred at chlorhexidine concentrations less than 0.013% and at povidone-iodine concentrations less than 0.5% (p less than 0.05). Significant S. aureus survival (p less than 0.05) was noted at chlorhexidine concentrations less than 0.05% and povidone-iodine concentrations less than 1.0%. These data showed that all bactericidal concentrations of chlorhexidine diacetate and povidone-iodine were lethal to canine embryonic fibroblasts in vitro, whereas non-lethal concentrations allowed significant bacterial survival.     

Effects of Chlorhexidine Diacetate and Povidone-lodine on Wound Healing in Dogs

To correlate the results of an in vitro study with clinical response, the effects of 0.005 and 0.05% chlorhexidine diacetate and 0.1 and 1.0% povidone-iodine concentrations on wound healing were evaluated in five beagle dogs. Full-thickness skin wounds (2 X 2 cm) were irrigated once daily for 14 days with the antiseptic solutions or physiologic-buffered saline. Chlorhexidine diacetate 0.05% had significantly more bactericidal activity than povidone-iodine and saline, and both chlorhexidine diacetate concentrations had residual effects 6 hours after irrigation. Neither povidone-iodine nor saline had significant bactericidal activity. The percentages of unhealed wound area and wound contraction were calculated 7, 14, and 21 days after wounding. Healed wound area and contraction were similar in wounds treated with chlorhexidine diacetate and povidone-iodine. However, wounds treated with chlorhexidine diacetate had more healed wound area on days 7 and 14 and more contraction on days 7, 14, and 21 than saline-treated wounds. At the concentrations tested, chlorhexidine diacetate irrigations provided bactericidal activity and were more beneficial to wound healing than irrigations with saline alone. These results suggest that concentrations of chlorhexidine diacetate which are cytotoxic to tissue culture fibroblasts in vitro do not interfere with wound healing in vivo.     

Evaluation of surgical scrub and antiseptic solutions for surgical preparation of canine paws

The purpose of the prospective study reported here was to evaluate surgical preparation of canine paws. Three combinations of surgical scrub solutions and antiseptic solutions were used: (1) 7.5% povidone-iodine scrub/10% povidone-iodine solution; (2) 2% chlorhexidine acetate scrub/2% chlorhexidine diacetate solution; and (3) tincture of green soap/70% isopropyl alcohol. The control was warm (38 to 42 C) tap water. Four microbial colony counts were used to evaluate surgical preparation of 4 paws of 8 dogs. Specimens were obtained from the paws for a baseline microbial flora count. After surgical scrub was performed, additional specimens were obtained for bacteriologic culturing. Antiseptic was applied followed by collection of another specimen for bacteriologic culturing. A final specimen was obtained following a 24-hour period under a sterile occlusive bandage. The 3 scrub solutions and the tap water control resulted in lower colony counts following scrubbing of the paws; however, only the 3 antiseptic solutions resulted in further colony count reduction after their application. Evaluation of residual colony counts isolated from specimens taken after a 24-hour period under a sterile occlusive bandage revealed chlorhexidine and povidone-iodine scrub/antiseptic combinations to be similar in antibacterial activity, with significantly (P less than or equal to 0.05) lower colony counts than those from specimens of paws treated with either the tincture of green soap/isopropyl alcohol combination or the tap water control. The lack of a significant difference between the bacterial counts immediately after surgical preparation with povidone-iodine and chlorhexidine and their respective 24-hour residual counts, indicated no particular advantage to surgical preparation and occlusive bandaging 24 hours prior to surgery.(ABSTRACT TRUNCATED AT 250 WORDS)     

Effects of 0.05% Chlorhexidine Lavage on the Tarsocrural Joints of Horses

In six horses, a 0.05% solution of chlorhexidine diacetate was used to lavage one tarsocrural joint; the contralateral control joint was lavaged with lactated Ringer's solution. Horses were evaluated daily for lameness. Synovial fluid samples were collected on days 1, 4, and 8 for determination of protein concentration, total and differential leukocyte counts, and mucin clot formation. After death on day 8, synovium and osteochondral samples were collected from the tarsocrural joints for examination of morphology and proteoglycan staining. Lavage with chlorhexidine solution caused lameness that was reduced but still evident at day 8. Synovial protein concentration was significantly increased by chlorhexidine lavage; the greatest increase occurred on day 1. Joint lavage increased synovial leukocyte counts on day 1, primarily by increasing polymorphonuclear (PMN) cell counts. Although total synovial leukocyte counts returned to normal by day 4, PMN cell counts remained elevated through day 8; PMN cell counts for chlorhexidine-lavaged joints were typically twice that of control joints. Chlorhexidine lavage caused synovial ulceration, inflammation, and abundant fibrin accumulation. Consistent differences in proteoglycan staining were not detected between control and chlorhexidine-lavaged joints. Joint lavage with 0.05% chlorhexidine diacetate, the lowest known bactericidal concentration, is not recommended for equine joints.     

Comparison of Three Skin Preparation Techniques in the Dog Part 1: Experimental Trial

Premeasured, clipped areas of skin on both sides of 30 adult dogs were prepared with povidone-iodine (PI), chlorhexidine gluconate (CG) with a saline rinse, or 4% CG with a 70% isopropyl alcohol rinse. Skin bacteria were quantified with Replicating Organism Detection and Counting (RODAC) plates and cultured for identification before, immediately after, and 1 hour after skin preparation. The percentages of bacterial reduction immediately and at hour 1 and the percentages of negative cultures, cultures with more than five colony-forming units (CFUs), and skin reactions were analyzed by analysis of variance and chi-square. The percentage of reduction in skin bacteria for all techniques was significant and comparable with that reported in humans. There were no significant differences between PI and CG results except that acute contact dermatitis was observed more frequently after skin preparation with PI. The authors conclude that for similar application times, PI and 4% CG rinsed with saline or 70% isopropyl alcohol are equally effective for up to 1 hour in the preoperative skin preparation of dogs.     

Comparison of Three Skin Preparation Techniques Part 2: Clinical Trial in 100 Dogs

The skin of 100 dogs undergoing clean or clean-contaminated surgical procedures was prepared with povidone-iodine (PI) or 4% chlorhexidine gluconate (CG) with saline or 70% isopropyl alcohol rinse. Skin bacteria at the incision site were quantified with Replication Organism Detection and Counting (RODAC) plates immediately before and after skin preparation in the preparation room, in the operating room, and postoperatively. The percentage of bacterial reduction, negative cultures, cultures with more than five colony-forming units, and skin reactions for each technique were calculated for each sample period and analyzed with the analysis of variance and Fischer tests. The percentage of bacterial reduction for all techniques was significant and comparable with results of a previous experimental study. There were no significant differences in percentages of bacterial reduction between PI and the CG techniques for surgical times up to 8 hours. There were fewer negative cultures and more cultures with high bacterial counts with PI than with CG and saline after the cleansing scrub. There were fewer negative cultures after surgery with CG and alcohol than with the other two techniques. Duration of the surgical procedure did not significantly affect the culture results. Significantly more skin reactions occurred with PI. The authors conclude that PI and 4% CG with a saline rinse are equally effective in antimicrobial efficacy under clinical conditions. However, 4% CG with a 70% isopropyl alcohol rinse may be inferior in residual antimicrobial activity.     

Effects of Four Preparations of 0.05% Chlorhexidine Diacetate on Wound Healing in Dogs

Four chlorhexidine diacetate (CHD) antiseptic wound lavage preparations were evaluated in vivo to determine their effects on second intention wound healing in the dog in vitro to determine their relative antibacterial activity against Staphylococcus intermedius. Chlorhexidine was diluted to 0.05% in sterile water, 0.9% sodium chloride, lactated Ringers solution (LRS), and LRS that was allowed to form a precipitate with CHD. Control solutions included sterile water and LRS. There were no significant differences in wound contraction or epithelialization. All 0.05% CHD preparations provided 100% bacterial kill.     

Response of pony peritoneum to four peritoneal lavage solutions

Peritoneal lavage was performed on ponies to determine the effect on peritoneal surfaces. Lavage solution (20 L) was introduced into each pony's peritoneal cavity through catheters placed in the paralumbar fossa, and the solution was removed by drainage from the ventral portion of the abdomen. Six ponies each were lavaged with sterile saline (0.9% NaCl) solution, sterile saline solution containing 5 X 10(6) U of potassium penicillin and 3 g of neomycin or povidone-iodine diluted to 3% by volume with sterile saline solution, and 3 ponies were lavaged with povidone-iodine diluted to 10% with sterile saline solution. Peritoneal lavage catheters were inserted in 3 control ponies, but lavage fluids were not administered. Peritoneal fluid specimens were collected at 6, 24, 48, and 96 hours after lavage. Nucleated cell counts, RBC counts, total protein determinations, and cytologic analysis were performed. The ponies were euthanatized at 96 hours, and representative sections of the peritoneum were examined. Lavage with saline solution and saline solution with antibiotics induced a mild, transient inflammatory response in the peritoneal fluid, with minimal or no changes observed at necropsy. Solutions containing povidone-iodine induced chemical peritonitis, which was severe in ponies lavaged with 10% povidone-iodine solution. Peritoneal lavage with povidone-iodine solutions as dilute as 3% cannot be accomplished without causing inflammation of peritoneal surfaces.     

Effect of antimicrobial solution lavage on the palmar digital tendon sheath in horses

Sixteen horses were allotted to 4 groups of 4 horses each to evaluate the effect of tendon sheath lavage with 4 solutions (balanced electrolyte solution, 0.1% povidone-iodine, 0.5% povidone-iodine, and 0.5% chlorhexidine). The synovitis caused by 0.1% povidone-iodine lavage was not appreciably worse than that caused by balanced electrolyte solution lavage, but the 0.5% povidone-iodine and chlorhexidine lavages caused severe synovitis, and, therefore, should not be used for tendon sheath lavage.     

Effect of Four Antimicrobial Lavage Solutions on the Tarsocrural Joint of Horses

Three concentrations of povidone-iodine (0.1% w/v, 0.2% w/v, 0.5% w/v) and one concentration of chlorhexidine (0.5% w/v) were selected as antimicrobial joint lavage solutions. Through-and-through joint lavage was performed with one of these antimicrobial solutions on a tarsocrural joint of 12 horses. The contralateral tarsocrural joints (control limbs) were lavaged with a balanced electrolyte solution (BES). The effect of the lavage solution on the joints was evaluated with respect to lameness, foot flight pattern, soreness to joint palpation, articular and periarticular enlargement, and synovial fluid composition on Day 1,4, and 8 postlavage. On Day 8 postlavage, all horses were euthanized and the tarsocrural joints were examined.
All solutions induced a synovitis. Based on clinical assessment, synovial fluid protein levels, color, clarity, mucin clot forming ability, gross appearance of the joint at necropsy, and synovial membrane histologic evaluation, a similar, mild, transient, synovitis was induced by the BES and 0.1% povidone-iodine (PI) solution. The 0.2% PI solution induced a more prolonged neutrophilic response and poorer mucin clot forming ability in the synovial fluid as compared to the BES.
The 0.5% PI and 0.5% chlorhexidine solutions produced severe lameness, soreness to joint palpation, and limb enlargement. The elevated synovial fluid total protein content persisted significantly longer (p < 0.05) than the corresponding control (BES) solution. Histologic evaluation of the synovial membrane confirmed the presence of a moderate to severe neutrophilic synovitis in these treatment groups.     

High pressure pulsatile lavage and high pressure syringe lavage in the treatment of contaminated wounds in dogs

The effectiveness of high pressure pulsatile lavage and syringe lavage in cleansing experimentally contaminated and infected wounds was studied. Such treatment significantly (P < 0.05) lowered the bacterial counts in contaminated wounds, but high pressure pulsatile lavage caused extensive damage to tissue and was considered to be unsuitable for treatment of wounds in the loose skin areas of the dog. Both pulsatile and syringe lavage caused significant (P < 0.05), but transient, reduction in the bacterial counts in infected wounds but both methods failed to lower the wound bacterial counts to 10⁵ or less. Successive wound biopsies for quantitative bacterial analysis showed that syringe lavage with povidone-iodine caused greater reduction in bacterial numbers than did pulsatile lavage with povidone-iodine. Combination of high pressure syringe lavage with povidone-iodine and systemic penicillin-streptomycin given at the time of lavage sufficiently reduced the bacterial numbers to allow for safe closure of the wounds.     

Comparison of four protocols for preoperative preparation in cattle

This study was designed to evaluate 4 preoperative skin preparations, that is, more specifically, to compare the efficacy of chlorhexidine gluconate (CG) and povidone-iodine (PI), as well as 2 hair removal techniques (clipper alone or clipper followed by razor) for preoperative skin preparation in cattle. The 4 protocols resulted in a significant decrease in the number of bacterial colony-forming units (cfu). Group 4 (clipping + shaving + CG) had a significantly lower number of preoperative cfu per gel plate compared with groups 1 (clipping + PI) and 3 (clipping + shaving + PI). Skin reaction frequency was significantly higher in groups 3 and 4 (47.8% for both protocols) than in groups 1 and 2 (clipping + PI or CG) (8.7% for both). Wound infection frequency was 4.3% (4/92) and no significant difference was observed between the 4 treatment groups. The 4 protocols tested were equivalent as to efficacy and satisfactorily decreased skin microflora. Clipping alone was shown to be preferable to clipping plus shaving as a method of hair removal in cattle, with fewer skin reactions and no more wound infections.     

双乙酸钠在肉鸭日粮中的添饲效果试验

试验以樱桃谷肉鸭为研究对象,进行了不同双乙酸钠添加水平的饲养对比试验,试验分三组,A为基础日粮,B、C分别添加0.2%、0.4%的双乙酸钠。结果表明:日粮中添加双乙酸钠对肉鸭日增重、瘦肉率有显著影响(P<0.05);对肉鸭的饲料转化率有极显著影响(P<0.01);对肉鸭的成活率和屠宰率、皮脂率无显著影响(P>0.05),B组日增重比A组提高了8.65%(P<0.05),比C组提高了5.45%(P>0.05);B组料肉比较A组降低了11.52%(P<0.01),较C组降低了3.29%(P<0.05);B组的成活率与A组、C组间差异不显著(P>0.05);B组每只肉鸭分别比A、C组多盈利1.04元、1.06元。     

Antibacterial activity of dilute povidone-iodine solutions used for ocular surface disinfection in dogs

Bacterial cultures of specimens from healthy canine eyelids and ocular surfaces were found to demonstrate bacterial growth in 69.7% (53/76) of the eyes sampled. Organisms most commonly isolated included: Staphylococcus aureus, alpha-hemolytic Streptococcus sp, S epidermidis, and Escherichia coli. Evaluation of dilute povidone-iodine solutions for effectiveness as ocular surface disinfectants was conducted. Bacterial growth initially detected in 32 of 46 eyes was not detected after disinfection with a 2-minute scrub and 2-minute soaking procedure, using 1:2, 1:10, or 1:50 dilutions of a povidone-iodine solution that contained 1% available iodine. The eyelid and ocular surfaces of 16 eyes were disinfected with 1:100 povidone-iodine solution. Bacterial growth initially present in 10 of 16 eyes was present in 1 eye after disinfection and consisted of a single colony of E coli. After eyes were disinfected with 1:10, 1:50, or 1:100 povidone-iodine solutions, there was no evidence of corneal epithelial edema or sloughing. In 15 eyes subjected to disinfection with the 1:2 dilution, one instance of epithelial corneal edema was noticed. A 1:50 dilution of povidone-iodine is recommended as an ocular surface disinfectant for use in presurgical situations.     

Comparison of three preoperative skin preparation techniques in ponies

The purpose of this study was to evaluate the efficacy of 3 preoperative skin preparations (povidone‐iodine [PI] removed with 70% isopropanol, and 4% chlorhexidine gluconate [CG] removed with either 70% isopropanol [CA] or sterile saline solution [CS]) in ponies. Eighteen ponies were randomly assigned to one of the 3 preoperative skin preparation groups. The skin of ponies was aseptically prepared with PI removed with alcohol, or 4% CG removed with either alcohol or sterile saline solution. The antimicrobial efficacy of each skin preparation technique was assessed quantitatively by culturing for surface bacteria with replicating organism detection and counting plates. The percentage of negative cultures, the percentage of cultures with >5 colony forming units (CFU) and the percentage bacterial reduction after the cleansing scrub, the sterile scrub, and the surgical procedures were compared. There was a significant difference between CG and PI in percentage bacterial reduction after the cleansing scrub, but no significant difference at any other time (sterile scrub, post operative skin sampling). In a comparison of the number of negative microbial cultures at each sampling point, there were no significant differences among the 3 skin preparation techniques. There were no significant differences among the treatment groups comparing the number of cultures with a high colony count (>5 CFU) after the cleansing scrub. Skin preparation with CS and PI resulted in significantly fewer cultures with >5 CFU after the sterile scrub than CA. Post operatively, CA had a higher number of samples with >5 CFU than CS and PI. PI removed with alcohol and 4% CS are equally effective in the reduction of skin bacteria after a sterile skin scrub in the operating room; however, CG was more effective in reducing bacterial numbers after the cleansing scrub. The number of cultures with high bacterial counts was greater post operatively, when alcohol was used as a rinse with chlorhexidine. In cases where a sterile scrub is not performed following a cleansing scrub, CG may be a better choice than PI. CA should not be used as a presurgical skin preparation method in ponies.     

复方氟康唑软膏临床药效试验

为研究复方氟康唑软膏对犬耳道真菌、细菌感染的治疗效果,发犬为试验用动物,采用人工感染和自然感染犬,以耳肤灵为对照,进行42d的临床药效研究。人工感染临床试验结果显示,复方氟康唑软膏和耳肤灵对犬耳道感染均有较好的治疗效果,两者无显著性差异(P>0.05);复方氟康唑软膏推荐剂量组和加倍剂量组对耳道感染的治愈率+显效率均为90%,且与空白对照组相比,差异极显著(P<0.01)。自然感染临床药效试验数据表明,耳肤灵和复方氟康唑软膏对真菌、细菌引起的耳道感染的治愈率+显效率均大于90%,且差异不显著(P>0.05),再次验证了复方氟康唑软膏推荐剂量对犬耳道细菌、真菌感染的治疗效果。表明复方氟康唑软膏推荐剂量(0.5mL/耳,1次/d,连续给药4周)可用于犬耳道细菌、真菌感染的治疗,治疗效果较好,可以在临床中推广使用。     

Comparison of the 5.8S rRNA gene and internal transcribed spacer regions of trichomonadid protozoa recovered from the bovine preputial cavity.

Sequence analysis of the 5.8S rRNA gene and the internal transcribed spacer regions (ITSRs) was used to compare trichomonadid protozoa (n = 39) of varying morphologies isolated from the bovine preputial cavity. A multiple sequence alignment was performed with bovine isolate sequences and other trichomonadid protozoa sequences available in GenBank. As a group, Tritrichomonasfoetus isolates (n = 7) had nearly complete homology. A similarity matrix showed low homology between the T. foetus isolates and other trichomonads recovered from cattle (<70%). Two clusters of trichomonads other than T. foetus were identified. Eighteen isolates comprised 1 group. These isolates shared >99% homology among themselves and with Pentatrichomonas hominis. The other non-T. foetus cluster (n = 14) did not exhibit a high degree of homology (<87%) with other bovine isolates or any of the trichomonad sequences available in GenBank. The sequence homology among isolates in that cluster was >99%, except for 1 isolate that varied from the others in both ITSRs (approximately 2% dissimilarity). Sequence analysis of the 5.8S rRNA gene and ITSRs was useful for comparing trichomonadid protozoa isolated from the bovine preputial cavity and demonstrated that 2 distinct types of trichomonads constituted the non-T. foetus isolates recovered from the bovine preputial cavity.     

Effects of Chlorhexidine Gluconate and Chlorous Acid-Chlorine Dioxide on Equine Fibroblasts and Staphylococcus aureus

Equine fibroblasts and Staphylococcus aureus were exposed for 30 minutes to six dilutions of chlorhexidine gluconate, a chlorous acid-chlorine dioxide irrigation solution, a chlorous acid-chlorine dioxide disinfectant, and phosphate buffered saline controls. Cell viability was determined by trypsinizing the cells, staining them with trypan blue, and counting cells that did not take the stain. All fibroblasts were killed when exposed to 1.0% and 0.5% chlorhexidine. The survival rate of fibroblasts increased linearly with decreasing concentrations of chlorhexidine gluconate, with a peak survival of 50% at 0.005% chlorhexidine. The chlorous acid-chlorine dioxide irrigation solution was the least toxic to fibroblasts, with survival rates equivalent to those of controls. The chlorous acid-chlorine dioxide disinfectant was 100% cytotoxic even when diluted 1:1 with phosphate buffered saline. S. aureus growth was inhibited by 1.0% and 0.5% chlorhexidine gluconate; concentrations of 0.05%, 0.01%, and 0.005% did not differ from sterile water controls. The chlorous acid-chlorine dioxide irrigation solution did not inhibit growth of S. aureus in brain-heart infusion broth. The chlorous acid-chlorine dioxide disinfectant inhibited growth of S. aureus.     

A Comparison of Antimicrobial Efficacy and Tissue Reaction of Four Antiseptics on Canine Wounds

Three concentrations of povidone iodine, chlorhexidine, benzalkonium chloride, and a pluronic polyol were tested for antimicrobial efficacy and tissue reaction in wounds inoculated with Staphyloccocus aureus .
Five paired incisions were created on the back of each of 16 dogs in four treatment groups. Five wounds were inoculated with beta-hemolytic, coagulase positive S. aureus (10⁹ organisms); the contralateral wounds were not inoculated. After two hours, three wounds on each side were irrigated with antiseptic in low, medium, or high concentrations; another was irrigated with physiological saline; and one was left untreated. One hundred ml of solution was applied in each instance with a syringe and needle at approximately 8 psi.
Tissue bacteria were quantitated immediately following irrigation. The wounds were sutured with 5/0 monofilament stainless steel and bandaged. After 48 hours, the dogs were euthanized and signs of wound infection were recorded. Wound bacteria were quantitated again, and sections of the wounds were removed for examination by light microscopy.
Antiseptics were significantly effective (p < 0.01) in reducing the bacterial population in contaminated wounds when compared to control wounds on the same dog. High antiseptic concentrations were more effective than low concentrations. At 48 hours, the bacterial population was significantly lower in wounds treated with chlorhexidine or benzalkonium chloride compared with povidone iodine or pluronic polyol. The antiseptics caused no significant difference (p < 0.01) in tissue reaction in noninoculated wounds. Chlorhexidine (0.5 or 1.0%) was the most effective antiseptic under the conditions used.     

Prevalence of Histophilus ovis and Actinobacillus seminis in the genital tract of sheep

Histophilus ovis and Actinobacillus seminis were isolated from the preputial cavity of 6-month-old rams and the vagina of 6-month-old ewes at a substantially higher rate than that in mature (greater than 2 years old) rams and ewes. These organisms appeared to be a transitory component of the ovine genital flora, the prevalence of which was associated with age regardless of gender. Additional evaluation of the recoverability of H ovis and A seminis from the preputial cavity of rams from birth to 1 year of age indicated that the isolation rate from rams and predominance of the organisms in the preputial cavity differed greatly over this age period. These organisms were not recoverable until ram lambs were 12 weeks of age and were most prevalent at 20 weeks of age, after which recoverability of H ovis and A seminis from the preputial cavity steadily decreased, continuing through the time of the last evaluation at 1 year of age. The time period with which these organisms can be isolated from the preputial cavity is closely correlated with the time period when epididymitis associated with these organisms develops, and may be an important factor in the pathogenesis of epididymitis.