Evaluation of Surgical Scrub Methods for Large Animal Surgeons

Objective— The objective of this study was to determine the effectiveness of a 5–minute surgical scrub using either a one-brush or a two-brush technique in clean and dirty surgical procedures, and to compare the efficacy of povidone iodine with chlorhexidine as surgical scrub solutions. Study Design— Prospective clinical trial. Methods— Nine veterinarians scrubbed their hands on eight separate occasions using either povidone iodine or chlorhexidine gluconate. A 5-minute scrub and either a one-brush or two-brush technique used in both clean and dirty operations were evaluated by taking glove juice samples before scrubbing, immediately after scrubbing, and 30, 60, 90, and 120 minutes after scrubbing. Glove juice samples were cultured and the colonies were counted. Percent reductions of bacterial forming units were calculated for all eight scrub procedures. Results— All scrub procedures provided an adequate percent reduction in colony forming units (CFU) during the 2–hour sampling period. The number of CFU immediately after scrubbing were significantly lower than prescrub. At 120 minutes, there were significantly fewer CFUs than presecrub, but there were more than immediately after scrubbing. No significant difference in reduction in CFUs were detected between one-brush and two-brush techniques. Both chlorhexidine and povidone iodine scrub solutions adequately reduced bacterial colony counts for 120 minutes after scrubbing regardless of the amount of contamination before skin preparation. Conclusions— Bacterial counts after a hand scrub procedure using a one-brush technique were not significantly different than after a procedure that used a two-brush technique. Povidone iodine and chlorhexidine are equally effectively in decreasing bacterial numbers on the skin, given a variety of contamination levels present before the scrub procedure. Clinical Relevance— Surgeons may use either chlorhexidine or povidone iodine for antiseptic preparation of their hands before surgery. A two-brush technique is not necessary.     

Comparison of an alcohol-based hand rub and water-based chlorhexidine gluconate scrub technique for hand antisepsis prior to elective surgery in horses

This prospective clinical study evaluates the effectiveness of an alcohol-based hand rub (Avagard™) for pre-surgical hand antisepsis in an equine hospital and compares it with traditional scrubbing technique using 4% chlorhexidine gluconate sponges and water. Prior to elective surgery, 3 board-certified surgeons were randomly assigned to hand antisepsis with either technique. Culture samples of each hand were taken at 4 times: before and after neutral soap hand wash, after scrub or rubbing technique, and after surgery. There was no significant difference in mean bacterial colony forming units between scrub and rub techniques over the 3 time periods (P = 0.6), controlling for initial counts. One horse from the scrub group had a skin incision infection following stifle arthroscopy; this was resolved with medical treatment. The alcohol-based hand rub is equivalent in efficacy for pre-surgical hand antisepsis to traditional water-based scrubs in an equine hospital setting.     

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)     

Comparison of Two Methods for Presurgical Disinfection of the Equine Hoof

OBJECTIVES: To determine for equine hooves the normal resident aerobic bacterial population and the efficacy of 2 methods of disinfection. Study Design-Measurement of total bacterial, gram-positive bacterial, and gram-negative bacterial surface populations from the frog, sole, and hoof wall after each step of 2 different preoperative surgical disinfection techniques. ANIMALS: Six adult horses. METHODS: Hoof wall, sole, and frog samples were collected for quantitative bacteriology before, during, and after 2 multistep antiseptic preparation techniques: Method A-6-minute scrub with povidone-iodine soap, followed by 24-hour submersion in povidone-iodine solution-soaked cotton; and Method B-initial removal of superficial layer of hoof capsule before completing Method A disinfection procedures. RESULTS: Removal of the superficial hoof layer, application of the povidone iodine scrub, and completion of the povidone-iodine soak all significantly (P < .0008) decreased total bacterial numbers. Method B had significantly lower bacterial counts than method A at each consecutive step. Final total bacterial counts remained greater than 10(5) bacteria per gram of tissue regardless of preparation method. CONCLUSIONS: The hoof surface hosts a broad spectrum of aerobic gram-positive and -negative bacteria, many of which are potential pathogens. Bacterial numbers can be significantly reduced by removal of the superficial hoof surface, by application of a povidone-iodine scrub, and by use of a 24-hour povidone-iodine soak. However, bacterial populations >10(5) g per tissue persist after these disinfection procedures. CLINICAL RELEVANCE: Regardless of the preparation methods used in this study, bacterial populations capable of inducing wound infection remain on the hoof capsule.     

Treatment of canine hip dysplasia: a review.

This article discusses the treatment approaches and recommendations for canine hip dysplasia. A search of the literature database MEDLINE (1969-1994) was conducted and relevant journal articles regarding the medical and surgical treatment of hip dysplasia were selected and reviewed. Dysplastic dogs can be divided, for treatment purposes, into those with no or minimal osteoarthrosis, and those with moderate to severe osteoarthrosis. In young animals with joint laxity and pain, but with no or minimal radiographic evidence of osteoarthrosis, the treatment approach is controversial. Conservative management may be effective in the short term, but progressive development of osteoarthrosis occurs and clinical signs may manifest at an older age. Options for surgical treatments in these young dogs include pectineal myectomy, lengthening of the femoral neck, and corrective osteotomies. Corrective osteotomies are advocated to reestablish joint congruency and prevent development of osteoarthrosis. In the mature osteoarthritic dog, effective conservative management depends on the severity of the degenerative joint disease. Proposed surgical treatments for clinically debilitating hip dysplasia include biocompatible osteoconductive/shelf arthroplasty; femoral head and neck excision arthroplasty, with or without muscle sling interposition; and total hip replacement. Although research directly comparing the salvage procedures has not been reported, studies suggest that total hip replacement is more effective in returning large dogs to full functional weight bearing.     

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.     

Evaluation of leukapheresis and thrombocytapheresis in the horse

Continuous-flow centrifugation leukapheresis techniques were used to collect 300-ml volumes of leukocyte-rich plasma from 5 nonmedicated horses and from 5 corticosteroid-stimulated horses. White blood cell counts and differential counts were performed on the horses before (base line) and up to 48 hours after leukapheresis. Systemic administration of hydrocortisone increased numbers of total WBC and neutrophils and improved harvest of these cells. Nonmedicated horses had a mean yield of 3.38 X 10(10) leukocytes in the 300-ml volume. Stimulated horses yielded a mean of 6.88 X 10(10) leukocytes. After leukapheresis, WBC counts decreased a mean of 38% in nonstimulated horses and decreased a mean of 30% in stimulated horses. By 6 hours after leukapheresis, circulating WBC counts of horses in both groups had returned to preleukapheresis values. The relationship between neutrophil yield and the 4 variables (preleukapheresis WBC count, preleukapheresis neutrophil count, preleukapheresis lymphocyte count, and the PCV of the leukocyte-rich plasma) were examined, using simple (pair-wise) correlation and multiple linear regression. A significant positive correlation was found between neutrophil yield and preleukapheresis WBC and neutrophil counts. Because sodium citrate was used in the collection system to prevent extracorporeal blood coagulation, ionized and total serum calcium concentrations were monitored before and after leukapheresis. Although total serum calcium concentrations remained unchanged, ionized calcium concentrations decreased approximately 33% from base-line values during the 2-hour leukapheresis procedures. Occasional mild muscle fasciculations were the only adverse clinical signs of citrate toxicity exhibited by the horses.(ABSTRACT TRUNCATED AT 250 WORDS)     

A Clinical Study of Canine Total Hip Arthroplasty

Our intent was to evaluate clinically total hip arthroplasty in the dog using the same procedures and prostheses. We replaced 20 hips in 15 dogs of both sexes and various breeds, weights, and ages. All animals exhibited pain caused by degenerative joint disease of one or both coxofemoral joints. Using Amstutz's system of grading at a minimum of 12 months post-operatively, none of the hips replaced were graded poor, 12.5% were graded fair, 12.5% good, and 75% excellent. Loosening of prosthetic components was by far the most common problem encountered in this study. Luxations were virtually eliminated by the use of the Richards Canine II total hip replacement system. Infections were found to be directly proportional to surgery time, number of operations performed, or both. Component failure occurred only when the size of the prosthesis was not properly matched to the size of the dog. Although these problems still must be overcome, we conclude that total hip arthroplasty is a valuable adjunct to veterinary medicine in selected cases.     

Evaluation of otoscope cone cleaning and disinfection procedures commonly used in veterinary medical practices: a pilot study

The objective of this study was to evaluate the relative efficacy of otoscope cone cleaning and disinfection methods commonly used in veterinary practices. Using sterile technique, 60 new gas-sterilized 4-mm otoscope cones were inoculated with a broth culture of 1.5 billion Pseudomonas aeruginosa bacteria per mL then allowed to dry for 10 min. Six study groups of 10 cones each were created. Group 1 served as positive control and received no cleaning or disinfection. Group 2 cones were wiped with sterile cotton-tipped applicators and gauze then rinsed with water. Group 3 cones were wiped with 70% isopropyl alcohol. Group 4 cones were scrubbed in a speculum cleaner with Cetylcide II solution (Cetylite Industries, Inc., Pennsauken, NJ). Groups 5 and 6 cones were soaked for 20 min in Cetylcide II and chlorhexidine gluconate 2% solutions, respectively. Using sterile technique and after 10-15 min drying time, the cones were swabbed in a consistent pattern, and samples were submitted for quantitative culture. Culture results showed no growth from cones soaked in Cetylcide II or chlorhexidine solutions. Two of the 10 cones wiped with alcohol, 3/10 cones wiped then rinsed with water, and 3/10 cones scrubbed with the speculum cleaner showed growth of P. aeruginosa. All (10/10) cones in the control group showed heavy growth of P. aeruginosa. These results show that P. aeruginosa can survive on otoscope cones cleaned and disinfected by several commonly used methods. Further study is needed to determine practical and optimal cleaning and disinfection methods for otoscope cones.     

Ranging behaviour and duration of survival of wild brushtail possums (Trichosurus vulpecula) infected with Mycobacterium bovis

AIM: To quantify the duration of survival of possums (Trichosurus vulpecula) infected with Mycobacterium bovis, and identify aspects of their behaviour which may influence the likelihood of disease transmission to domestic stock or wildlife. METHODS: Capture and den locations of 14 naturally infected tuberculous possums, eight possums experimentally infected with M. bovis and eight non-infected possums were recorded between May 1998 and February 2000 at a study site near Castlepoint on the Wairarapa coast of the North Island in New Zealand. Denning behaviour was observed weekly using radiotelemetry, and possums were captured, examined and released bi-monthly. Data were used to estimate survival period; create denning, activity, and total ranges; and to identify extended forays by possums as individuals and groups. RESULTS: Seventeen tuberculous possum carcasses were recovered, of which 14 (82%) were close to or within their activity range. Denning ranges were known for 10/17 possums that died. Four tuberculous possums were found dead within their denning range. Three possums made extended forays in the 3 weeks before death. Twelve possums were found dead in dense scrub, three in long grass in open woodland and two on pasture. Mean duration of survival of naturally infected possums following detection of clinical signs was 3.4 months (95% CI=2.1-5.4) and the instantaneous mortality rate was 0.293 per month (95% CI=0.184-0.470). Signs of disease were obvious for about 3 weeks prior to death. Tuberculous possums were commonly trapped on only part of the area where the total non-infected population was trapped. CONCLUSION: Most tuberculous possums died within their activity range and in scrub, representing a risk of transmission of M. bovis to wildlife and livestock that forage in scrub. Smaller proportions dying on pasture represent a less frequent, but highly visible risk. Tuberculous possums were clustered on the study site, and localised possum control operations would be more effective if focussed on such areas.     

Sequential clinical and synovial fluid changes associated with acute infectious arthritis in the horse

Infectious arthritis was induced experimentally in one tarsocrural joint of six horses by intra-articular injection of 1 ml Staphylococcus-saline suspension containing 9 x 10(4) to 3 x 10(6) organisms. The corresponding contralateral joint was injected with 1 ml of saline and served as a control. The progression of the induced infectious arthritis was assessed over a nine-day period by clinical examination and sequential synovial fluid analysis with pH and lactate measurements. Changes in synovial fluid were present before clinical signs of infectious arthritis were manifested. The diagnostic value of different synovial fluid parameters at various stages of infection was determined. Cellular changes initially preceded the biochemical changes. Total leucocyte counts showed a significant increase within 24 h (up to 100 x 10(9)/litre) with great variability in subsequent measurements. Neutrophilia over 90 per cent and pH under 6.9 were the most consistent findings in the infected synovia. Increased total protein was also significant and was progressive throughout the experiment. Serum and synovial glucose difference and synovial lactate had more diagnostic value in the acute stages than in the chronic stages. The control joints elicited an inflammatory response manifested by increased leucocyte count, moderate neutrophilia, slightly increased total protein concentration, and slightly decreased pH, but all reactions were minor in comparison to those in the infected joints.     

Evaluation of arthrocentesis site bacterial flora before and after 4 methods of preparation in horses with and without evidence of skin contamination

OBJECTIVE: To evaluate the effectiveness of four methods of povidone-iodine preparation on skin bacterial flora of arthrocentesis sites, in horses, with and without evidence of skin contamination. STUDY DESIGN: Prospective randomized study. ANIMALS: Twenty-four adult horses. METHODS: Horses were assigned to either the clean or contaminated group based on housing environment and visual evidence of contamination. Using a moist sterile swab, microbial culture samples were obtained from the skin over the distal interphalangeal joints immediately before and after preparation. Each site was aseptically prepared with 1 of 4 povidone-iodine techniques: 10-minutes scrub, 5-minutes scrub, three 30-second scrubs, or commercial one-step iodophor surgical solution. Colony forming units (CFUs) were determined for each sample, 24 hours after inoculation, on blood agar plates. RESULTS: Mean (+/-SD) pre-scrub CFUs/mL was significantly higher in the contaminated group (9588.33+/-1223.65) compared with the clean group (4489.00+/-3842.03) (P<.01). After preparation of the arthrocentesis sites, there were no significant differences in post-scrub CFUs/mL among the 10 minutes (mean clean, 46.00+/-64.36; mean contaminated, 28.67+/-18.04), 5 minutes (mean clean, 84.17+/-109.80; mean contaminated, 40.33+/-44.52), three 30 seconds povidone-iodine scrubs (mean clean, 95.50+/-172.29; mean contaminated, 46.67+/-56.94), or application of a commercial one-step iodophor surgical solution (mean clean, 102.17+/-161.78; mean contaminated 117.67+/-143.78); or between the clean (81.96+/-131.69) and contaminated groups (58.33+/-85.90) (P<.01). CONCLUSIONS: Preparation of the distal interphalangeal joint arthrocentesis site with each of these techniques significantly reduces the bacterial flora to a similar level for arthrocentesis in horses with and without evidence of skin contamination. Clinical Relevance- Aseptic preparation of the skin over the distal interphalangeal joint may be accomplished with any of these techniques.     

Bactericidal effects of ozone at nonspermicidal concentrations.

A study was conducted to assess the use of ozone (O3) to control pathogens or contaminants of concern to animal breeders and regulatory officials. In separate experiments, samples of fresh bovine semen and Pseudomonas aeruginosa, Escherichia coli, or Campylobacter fetus subsp. venerealis were diluted with antibiotic-free milk (10(6) sperm and 10(6) organisms/mL of diluted semen), exposed in the previous day to a constantly monitored level of 5, 10, 15, or 20 micrograms/mL of O3 for 3-5 min. After 10 min at 30 degrees C, sperm motility was assessed and the samples cooled to 5 degrees C. Two and 18 h after the beginning of cooling, aliquots of each semen sample were evaluated for motility and cultured for organisms. Reductions were observed in P. aeruginosa and E. coli colony counts of 2 logs, and in C. fetus of 5 logs, after exposure for 2 h to O3 at a concentration of 5 micrograms/mL that had a moderate effect on sperm motility (reduction of 20%). Fewer than 100 colonies, i.e., a 4 logs reduction of all bacteria, were counted after dilution with ozonized-treated milk at 20 micrograms/mL of O3. However, this concentration of O3 reduced sperm motility by 50% 10 min after dilution. The results of these experiments indicate that a concentration and exposure time to O3 can be selected to reduce P. aeruginosa, E. coli, and C. fetus in contaminated bull semen diluted with milk while having only minimal effects on sperm motility.     

Idiopathic brood disease syndrome and queen events as precursors of colony mortality in migratory beekeeping operations in the eastern United States

Using standard epidemiological methods, this study set out to quantify the risk associated with exposure to easily diagnosed factors on colony mortality and morbidity in three migratory beekeeping operations. Fifty-six percent of all colonies monitored during the 10-month period died. The relative risk (RR) that a colony would die over the short term (～50 days) was appreciably increased in colonies diagnosed with Idiopathic Brood Disease Syndrome (IBDS), a condition where brood of different ages appear molten on the bottom of cells (RR = 3.2), or with a “queen event” (e.g., evidence of queen replacement or failure; RR = 3.1). We also found that several risk factors—including the incidence of a poor brood pattern, chalkbood (CB), deformed wing virus (DWV), sacbrood virus (SBV), and exceeding the threshold of 5 Varroa mites per 100 bees—were differentially expressed in different beekeeping operations. Further, we found that a diagnosis of several factors were significantly more or less likely to be associated with a simultaneous diagnosis of another risk factor. These finding support the growing consensus that the causes of colony mortality are multiple and interrelated.     

Resistance to thiabendazole, fenbendazole and levamisole in Haemonchus and Trichostrongylus species in sheep on a Kenyan farm

Resistance to thiabendazole (TBZ), fenbendazole (FBZ) and levamisole (LVM) in naturally acquired gastrointestinal nematode parasites in sheep was investigated on a farm where anthelmintic resistance was suspected. This was measured by both the in vitro egg hatch assay, and reductions in faecal egg and worm counts in treated animals. In the egg hatch assay, nematode eggs were incubated in various concentrations of either TBZ or LVM. The level of resistance was expressed as the drug concentration inhibiting 50% of the eggs from hatching (LC50). The nematode population had LC50 values of 0.26 microgram ml-1 TBZ and 3.12 micrograms ml-1 LVM. In the faecal egg and worm count reduction test, naturally infected sheep were treated with either TBZ (88 mg kg-1), FBZ (10 mg kg-1) or LVM (15 mg kg-1). Faecal egg and total worm counts from these sheep were then compared with counts from untreated sheep. TBZ, FBZ and LVM failed to reduce the faecal egg counts and total worm counts by more than 90%. Based on the identification of larvae from faecal cultures, the most predominant nematode species in the resistant population were Haemonchus (62%) and Trichostrongylus (28%). TBZ reduced faecal egg counts for both species by less than 90%. FBZ and LVM also reduced Haemonchus spp. eggs by less than 90%. Other nematode species numbers did not satisfy criteria for the determination of efficacy.     

Treatment of fresh poultry carcases with emulsions of glycerol monocaprate (monocaprin) to reduce contamination with Campylobacter and psychrotrophic bacteria

1. A previous study has shown that emulsions of monocaprin in citrate lactate buffer at pH 4·1-4·3 are highly active in killing Campylobacter in water, where they reduce viable bacterial counts by more than 6 log(10) colony forming units (cfu) in 1 min at a concentration of 1·25 mM (0·03%). 2. The present study was carried out to evaluate whether monocaprin emulsions could be used to kill Campylobacter on raw poultry. 3. It was shown that immersion of naturally contaminated chicken legs in 20 mM (0·5%) monocaprin emulsion at pH 4·1 for 1 min at 20°C reduced the number of Campylobacter by 2·0 to 2·7 log(10) cfu. Pre-chill dipping of whole carcases into 20 mM monocaprin emulsion in the slaughterhouse also caused a significant reduction in Campylobacter contamination. 4. Immersion in monocaprin emulsions at pH 4·1 was also assessed as a means to reduce the number of psychrotrophic spoilage bacteria. There were lower psychrotrophic bacteria counts on treated chicken parts than on untreated controls after storage at 3°C for up to 14 d. 5. Immersion in emulsions of monocaprin, which is a natural lipid classified as GRAS, may be a feasible method to reduce the number of Campylobacter and spoilage bacteria on raw poultry. This method could reduce the risk of human exposure to Campylobacter, and at the same time increase the shelf-life of poultry products.     

Comparison of the bacterial flora of the duodenum in healthy cats and cats with signs of gastrointestinal tract disease

OBJECTIVE: To determine whether a colony environment predisposes healthy cats to high bacterial counts, including counts of obligate anaerobes, in the duodenum and whether increased numbers of bacteria could be found in the duodenum of cats with signs of chronic gastrointestinal tract disease. DESIGN: Prospective study. ANIMALS: 20 healthy control cats (10 from a colony environment and 10 pet cats) and 19 cats with a history of chronic gastrointestinal tract disease. PROCEDURE: Undiluted duodenal fluid was quantitatively and qualitatively assessed by bacteriologic culture under aerobic and anaerobic conditions. Serum concentrations of cobalamin and folate were also measured. RESULTS: Significant differences were not detected in the numbers of bacteria found in the duodenum of cats housed in a colony environment, compared with pet cats fed an identical diet prior to sampling. All healthy cats were, therefore, combined into 1 control group. Compared with healthy cats, cats with clinical signs of gastrointestinal tract disease had significantly lower counts of microaerophilic bacteria, whereas total, anaerobic, and aerobic bacterial counts were not significantly different. None of the cats with disease had total bacterial counts higher than expected from the range established in the control cats. Differences were not detected in regard to serum folate or cobalamin concentrations between diseased and healthy cats. CONCLUSIONS AND CLINICAL RELEVANCE: These findings indicated that healthy colony cats and pet cats have high numbers of bacteria in the duodenum, including high numbers of obligate anaerobes. Our findings also suggest that bacterial overgrowth in the small intestine is not a common clinical syndrome in cats with chronic nonobstructive gastrointestinal tract disease.     

Cryopreservation of the semen collected by electroejaculation from the Hokkaido brown bear (Ursus arctos yesoensis)

Four adult Hokkaido brown bears were used as semen donors, and semen characteristics were examined before freezing and after thawing. A total of 10 electroejaculates were diluted with Tris-egg yolk extender and cooled to 4 degrees C over 90 min. Spermatozoa were equilibrated with 4.7% glycerol for 80 min. Semen packed in 0.25 ml plastic straws were frozen with liquid nitrogen vapor. Percentages (mean +/- SD) of motile and live sperm were 96+/-2 and 86.5+/-7.2% before freezing, and 43+/-5 and 67.4+/-3.9% after thawing, respectively. Although the number of progressively motile sperm after thawing varied among samples (1.8+/-1.2 x 10(8) cells/ejaculate), frozen semen in the present study might serve for artificial insemination.     

An assay to quantitate the binding of Rhodococcus equi to macrophages.

A Rhodococcus equi radiobinding assay has been developed using organisms labeled with 3H-uracil. These labeled organisms resemble their unlabeled counterparts with respect to colony morphology, viability, and buoyant density. Bacteria routinely incorporate between 5 x 10(-3) and 5 x 10(-2) counts per minute per colony forming unit (cfu) which in this assay allows the detection of fewer than 0.2 cfu per macrophage. Once incorporated, greater than 90% of the label remains bacterial associated for at least 4 h postlabeling. The majority of the label is trichloroacetic acid precipitable, partitions into the aqueous phase following phenol/chloroform extraction and is ethanol precipitable. RNAse treatment of the ethanol precipitate abolishes label trichloroacetic acid precipitation. This radiolabeling technique has been used to quantitate the attachment of R. equi to both murine peritoneal and equine alveolar macrophages adherent to 13 mm glass coverslips. R. equi binding is dose dependent, saturable, and specific to macrophages. Further, binding is enhanced in the presence of fresh serum. Inhibition of radiolabeled bacterial binding can be obtained by competition with cold R. equi. This radiolabeled binding assay represents a crucial step in identifying the receptors on macrophages involved in the recognition of R. equi and may help to provide information on how macrophages recognize intracellular bacteria in general.