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The incidence and severity of disease associated with toxigenic Clostridium difficile have increased in hospitals in North America from the emergence of newer, more virulent strains. Toxigenic C. difficile has been isolated from food animals and retail meat with potential implications of transfer to human beings. The objective of the present study was to determine the prevalence of toxigenic C. difficile in chickens and retail poultry meat in Texas. Seven C. difficile isolates were detected in fecal samples of 300 (2.3%) broiler chickens. Three cultivation procedures were evaluated for isolation of C. difficile from poultry meat and detected 1/32 (3.1%), 2/32 (6.2%), and 4/32 (12.5%) for the three procedures, respectively. Chicken and poultry meat isolates were characterized as toxinotype V and pulsed-field gel electrophoresis gel type-NAP7 or NAP7-variant. Susceptibilities to 11 antimicrobial agents in the current study suggested somewhat reduced resistance than reported for other meat or animal toxinotype V isolates.

The incidence and severity of disease associated with toxigenic Clostridium difficile have increased in hospitals in North America from the emergence of newer, more virulent strains. Toxigenic C. difficile has been isolated from food animals and retail meat with potential implications of transfer to human beings. The objective of the present study was to determine the prevalence of C. difficile in pork from sausage manufacturing plants and retail meat in Texas. Twenty-three C. difficile isolates were detected from 243 meat samples (9.5%) from 3 sausage-manufacturing plants and 5 retail meat outlets from 2004 to 2009. Twenty-two isolates were positive for toxins A, B, and binary toxin, and were characterized as toxinotype V, PFGE type-NAP7, or "NAP7-variant." Susceptibilities to 11 antimicrobial agents in the current study were similar to those reported previously for toxinotype V isolates, although the results suggested somewhat reduced resistance than reported for other meat, animal, or human clinical toxinotype V isolates.

A continuous-flow porcine cecal bacterial culture has been used experimentally as treatment against enterotoxigenic Escherichia coli in weanling pigs. Periodically, the cultures must be started from frozen stock. Our results indicate that denaturing gradient gel electrophoresis can be applied as an indirect indication of culture similarity for each new batch generated from frozen stock.

Among the various quality elements which the Water Framework Directive requires should be monitored are macroalgae. One aspect of these is the presence, in transitional waters particularly, of large blooms of opportunistic macroalgae, such as Ulva and Enteromorpha. Within the United Kingdom (UK) and Republic of Ireland (RoI) there are currently no set ecological quality objectives or standards for macroalgae. Nor are there standard methods for monitoring macroalgal blooms, although various combinations of aerial photography, remote sensing and measurements on the ground are used. This paper attempts to set a logical framework for the prioritisation of sites for monitoring, the development of a tiered monitoring procedure and the derivation of thresholds for classification. Draft threshold limits for percentage cover and biomass of macroalgae have been derived from the literature. The importance of secondary effects and physico-chemical parameters is discussed.

Three experiments were conducted to evaluate the effects of different antibiotics in a milk-glucose semen extender on motility of equine sperm and elimination of bacteria following storage of extended semen in vitro. In Experiment 1, 7 antibiotics were compared: amikacin, gentamicin, streptomycin, potassium penicillin, sodium penicillin, ticarcillin, and polymixin B. In Experiment 2, 3 antibiotic treatments were compared: potassium penicillin G, amikacin, or a combination of potassium penicillin G and amikacin. In Experiment 3, 3 antibiotic treatments were compared: potassium penicillin G-amikacin, ceptiofur, and a combination of ticarcillin and clavulanic acid (Timentin). Control treatments (antibiotic-free extender) were included in each experiment. Six motility variables were evaluated: percentage of motile sperm; percentage of progressively-motile sperm; percentage of rapidly-motile sperm; mean curvilinear velocity; mean average path velocity; and mean straight-line velocity. In Experiment 1, mean percentages of motile, progressively motile and rapidly motile sperm were lower (P < 0.05) in semen exposed to polymixin B then in other treatments. Mean average-path velocity of sperm in extender containing polymixin B was lower (P < 0.05) than that of all other treatments, with exception of control or ticarcillin. Mean straight-line velocity of sperm in extender containing polymixin B was lower (P < 0.05) than that of all other treatments, with exception of control, streptomycin or ticarcillin. Semen samples containing gentamicin, amikacin, streptomycin, or potassium penicillin were more effective (P < 0.05) at eliminating bacterial growth than those samples containing polymixin B. Semen samples containing gentamicin were also more effective (P < 0.05) at eliminating bacterial growth than those samples containing ticarcillin or sodium penicillin. In Experiment 2, mean percentage of rapidly-motile sperm, and mean curvilinear, average-path, and straight-line velocities were greater (P < 0.05) for potassium penicillin-amikacin than values for all other treatments. In 2 of 3 stallions, an effect of treatment on percentage of motile sperm was detected (P < 0.05). For one stallion, mean motility of potassium penicillin-amikacin was greater (P < 0.05) than that of all other treatment groups. For another stallion, mean motility of the control was lower (P < 0.05) than that of the other treatments. Following storage, potassium penicillin (16/18 [89%]) or potassium penicillin-amikacin (17/19 [94%]) were more effective (P < 0.05) at controlling aerobic and anaerobic bacterial isolates in semen specimens than was amikacin (10/18 [56%]). In Experiment 3, a difference among treatment groups for motility variables was not detected (P < 0.05). No bacterial growth was recovered in antibiotic-treated semen, with exception of Micrococcus sp. (2 colonies) which were isolated from one semen specimen treated with ceptiofur.

Two defined competitive exclusion (CE) cultures (CF-I and CF-II) and a characterized CE culture (CF-III), which are composed of mixtures of nonpathogenic bacteria, were developed from anaerobic continuous-flow (CF) cultures that had been inoculated with cecal contents from adult chickens. After the primary CF cultures attained homeostasis, 13 bacteria (11 facultative anaerobes and 2 obligate anaerobes) representing 6 genera were isolated from CF-I; II bacteria (9 facultative anaerobes and 2 obligate anaerobes) representing 7 genera were isolated from CF-II; and 29 bacteria (15 facultative anaerobes and 14 obligate anaerobes) representing 14 genera were isolated from CF-III. Newly hatched chicks were treated orally with each primary CF culture; challenged on day 3 with Salmonella typhimurium; and cultured on day 10. Each culture significantly (p < 0.05) reduced salmonellae intestinal colonization and organ invasion. From the reconstituted CF-I and CF-II cultures, all organisms were isolated and their fermentation parameters and efficacy against salmonellae challenge were similar, if not identical, to the primary cultures. The CF-I and CF-II cultures satisfied the 5 requirements for defined CE cultures: 1) the primary CE culture must be efficacious; 2) all bacteria must be isolated and identified; 3) the fermentation parameters of the reconstituted CE culture must be similar to those of the primary culture; 4) all bacteria from the reconstituted culture must be isolated and identified; and 5) the efficacy of the reconstituted culture must be very similar to the primary culture. From these integrated studies, 3 mechanisms were demonstrated for preventing the enteric colonization of salmonellae in newly hatched chicks that were pretreated with CE cultures. First, the component organisms in the CE culture establish a normal enteric flora prior to salmonellae exposure. Second, the CE organisms compete with salmonellae for essential nutrients. Third, the CE organisms produce concentrations of volatile fatty acids at low pH levels that are bacteriostatic for salmonellae.

An 81-d-old continuous-flow (CF) culture of broiler cecal bacteria was maintained in a lactose-based broth. The culture had been previously proven effective against Salmonella colonization in young chicks, especially when the chicks were provided dietary lactose. Portions of the CF culture were batch-cultured in glucose- and lactose-based broths containing 14C-labeled lactose, glucose, galactose, or lactic acid to determine the effect of media carbohydrate on fermentation products. Acetic and propionic acids were the major 14C-labeled fermentation products. 14C-Carbohydrates were fermented to lactic acid and then to acetic and propionic acids. Distribution of radiolabeled fermentation products was effected by the broth carbohydrate and the time postinoculation.

The effect of treatment with a newly developed characterized continuous-flow (CCF) culture composed of 29 strains of cecal bacteria on salmonellae colonization was evaluated in commercially reared broiler chickens. Newly hatched chicks in three flocks were sprayed with CCF culture as they were placed in rearing houses and compared with untreated flocks on the same farm. Hatchery transport tray liners and rearing house feed, water, and little samples were cultured for the presence of salmonellae. Cecal samples were cultured after 3- and 6-wk growout. Skin-feather samples were cultured at 6-wk growout. Fifty-five percent of the transport tray liners were contaminated with salmonellae on the day of chick placement. At 3 wk, salmonellae serotypes present on the tray liners were widely distributed in the litter of the rearing houses. The results indicated that exposure to salmonellae occurred before culture treatment and continued in the rearing houses during the 6-wk growout period. Salmonellae cecal colonization was decreased (P < .05) in two of the treated flocks at 3 wk and diminished (P < .07) in the third treated flock compared with control flocks. At 6 wk, skin-feather contamination and cecal colonization were decreased (P < .05) in one of the treated flocks whereas no treatment effect occurred in two of the treated flocks compared with controls. The results clearly indicate the necessity of implementing integrated programs to control salmonellae in both the hatchery and rearing house environments. The CCF culture served to enhance salmonellae colonization resistance and may serve as a useful component of an integrated control program.

Glucose- and lactose-based media containing either 14C-labelled glucose, galactose, lactose or lactic acid were inoculated with anaerobic cultures of chicken caecal bacteria maintained for 121 d in a continuous-flow (CF) culture. The culture was previously shown to reduce Salmonella colonization in the caeca of chicks inoculated with the culture and the reduction was associated with increases in volatile fatty acids. The distributions of 14C were determined among the fermentation products, especially acetic, propionic and lactic acids. After 12 h of incubation and fermentation, variations were observed in the total amounts of 14C, from each 14C-labelled substrate, detected as acetic and propionic acids in the glucose- and lactose-based media, respectively: 92.9 and 89.4% of 14C-glucose, 77.0 and 44.0% of 14C-galactose, 0.0 and 76.9% of 14C-lactose, and 76.5 and 93.2% of lactic acid in the two media, respectively. The results from this study indicate that carbohydrate metabolism by the CF culture was a major source of acetic and propionic acids and that lactic acid was an important metabolic intermediate for the synthesis of the volatile fatty acids.

A continuous-flow culture system was used to isolate and maintain a mixed culture of cecal bacteria from adult broilers composed of 29 bacterial strains representing 10 genera. Broiler chicks were treated with the mixed culture in the drinking water on the day of hatch and challenged orally with 10(4) Salmonella typhimurium 2 d after treatment. The experiment was repeated in four separate trials using newly hatched chicks. The concentration of propionic acid and total volatile fatty acid (VFA) in the cecal contents was determined 2 d after treatment and at 10 d of age. Compared with controls, the number of treated chicks that were cecal culture-positive for Salmonella decreased (P < .01) in each of the trials. Additionally, the number of Salmonella in the cecal contents of the treated chicks at 10 d of age was decreased (P < .005) compared with controls in each trial. The decreased number of Salmonella in the cecal contents of the 10-d-old treated chicks was correlated with elevated concentrations of propionic acid (P < .05) and total VFA (P < .1) in the cecal contents of the treated chicks 2 d after treatment. The results indicated that VFA-producing bacteria present in the mixed culture became rapidly established in the ceca of the treated chicks and effectively increased resistance to S. typhimurium challenge.

A rumen fluid-based differential carbohydrate agar medium for enumerating chicken cecal carbohydrate-utilizing bacteria was tested. Eleven bacteria isolated from a continuous-flow culture that had been seeded with chicken cecal contents were inoculated on fifteen different media in an anaerobic environment. These media included various levels of rumen fluid (0%, 5%, 10%, 16%) and carbohydrate (glucose [GLU], galactose [GAL], lactose[LAC]). Viable cell recovery (colony-forming units [CFU]/direct cell counts × 100%) was higher for 16% rumen fluid-supplemented carbohydrate media than the lower concentrations of rumen fluid-supplemented carbohydrate media. In addition, average viable cell recovery of all 11 bacteria on differential carbohydrate media were significantly (P < 0.05) higher in galactose and lactose media than on the glucose media. Data indicated that 16% rumen-fluid based media can be used to enumerate anaerobic, lactose-fermenting subgroups of chicken cecal bacteria.

Leghorn chicks were treated with cultures of cecal bacteria from adult chickens by crop gavage, upper body spray, or vent lip application on the day of hatch. The chicks were challenged orally with 10(4) Salmonella enteritidis (SE) at 3 d of age and evaluated for SE cecal colonization at 10 d of age. The concentration of volatile fatty acids (VFA) in the cecal contents was determined on the day after culture treatment and at 10 d of age. Compared with controls, SE colonization was significantly decreased in each of the treatment groups. Vent lip application of a single .05-mL drop of cecal bacteria culture resulted in resistance against SE challenge comparable to crop gavage or spray treatment with .5 mL of culture. Resistance to SE challenge was directly associated with the concentrations of total VFA and propionic acid in the cecal contents of the treated chicks on the day after culture treatment. The results indicated that cecal bacteria from adult chickens that increase SE colonization resistance may rapidly become established in the ceca of newly hatched chicks following contact with the vent lips.

The effect of oral inoculation with a defined culture of cecal bacteria and provision of dietary lactose on colonization resistance to invasive Salmonella enteritidis was evaluated in leghorn chicks. A defined mixed culture composed of 11 indigenous cecal bacteria capable of utilizing lactose or lactose fermentation products as a primary carbon source was isolated and maintained in continuous-flow culture. Combined treatment with the defined culture and dietary lactose significantly decreased (P < 0.01) the number of Salmonella in the cecal contents and the number of Salmonella cecal-culture-positive chicks in four replicated trials. Additionally, Salmonella colonization of the spleen, liver, and cecal tonsils was significantly decreased (P < 0.05) in each of the trials. The results indicate that the resistance of leghorn chicks to S. enteritidis cecal and organ colonization is effectively increased by inoculation with a defined culture of cecal bacteria and provision of lactose in the diet.

The protective effect of cecal bacteria cultures on Salmonella enteritidis cecal colonization was evaluated. Competitive-exclusion cultures were administered by crop gavage, in first drinking water, by whole body spray, or encapsulated in alginate beads and provided in feed pans. Leghorn chicks were treated with cultures of cecal bacteria on the day of hatch and challenged orally with 10(4) S. enteritidis 2 days after treatment. Salmonella cecal colonization was evaluated 7 days after challenge. No Salmonella organisms were detected in the ceca of chicks treated with cecal cultures by crop gavage. Chicks treated with cecal cultures in the drinking water or by spray application showed comparable protection and significant decreases (P < 0.05) in the number of Salmonella in the cecal contents compared with untreated controls. The consumption of cecal bacteria encapsulated in alginate beads significantly decreased (P < 0.05) Salmonella cecal colonization compared with control treatment, but it provided less protection than the other treatment methods evaluated.

Experiments were conducted to examine the effect of a continuous-flow (CF) derived bacterial culture and of dietary lactose on colonization level by anaerobic bacteria in the ceca of broiler chicks. Cecal facultative and strictly anaerobic bacteria colony forming units (CFU), total volatile fatty acid (TVFA), lactic and propionic acid concentrations in 3-d-old chicks, and cecal log 10 Salmonella typhimurium colonization and propionic acid concentrations in 10-d-old chicks were measured. Treatment groups were control diet, 2% lactose diet, CF culture + control diet, and CF culture + 2% lactose diet. Groups inoculated with CF culture had decreased (P < 0.05) Salmonella in cecal contents at 10 d of age. The level of Salmonella protection was significantly (P < 0.05) correlated with increased cecal bacteria CFU, TVFA, and propionic acid concentrations in 3-d-old chicks. Results indicated that inoculating newly hatched chicks with CF culture containing cecal microflora originally obtained from adult chickens, facilitates early cecal colonization by native cecal microflora, and this is associated with protection against Salmonella .

The effect of dietary lactose and of cell concentration of a continuous-flow (CF) derived bacterial culture on Salmonella typhimurium cecal colonization in 10-d-old broiler chickens was examined. One-day-old chicks were provided 1) no CF culture and the control diet; 2) no culture and a 2% lactose diet; 3) CF culture (10(8) or 10(11) anaerobic cfu) and control diet; or 4) CF culture (10(8) or 10(11) anaerobic cfu) and 2 to 4% lactose diet. All groups were challenged orally with 10(4) S. typhimurium at 3 d of age. Salmonella typhimurium growth in cecal contents was significantly decreased (P < .05) at 10 d of age when 2% lactose was used in combination with CF culture containing 10(8) or 10(11) anaerobic cfu. The protection factor (log10 Salmonella control diet divided by log10 Salmonella treatment group) for these treatment groups was 2.49 and 9.26, respectively. Results indicate that birds treated with CF culture and maintained on 2% dietary lactose are protected against S. typhimurium colonization. Additionally, inoculating birds with CF culture containing a higher concentration of anaerobic colony-forming units resulted in a substantially higher protection factor.

Cecal anaerobic bacteria from adult broilers were cultured in media containing .25% glucose or .25% lactose. Media also contained either [14C]-labeled lactose, glucose, galactose, or lactic acid as metabolic tracers. Cultures were analyzed at 4, 8, and 12 h for pH, radiolabeled and unlabeled volatile fatty acids, and lactic acid. The pH values of cultures containing .25% lactose were significantly (P < .05) higher than the pH values of cultures containing .25% glucose. Lactose cultures reached their lowest pH more slowly than glucose cultures. Concentrations of unlabeled volatile fatty acids increased and lactic acid decreased during incubation of the cultures. Radiolabeled sugars and lactic acid were more readily metabolized to volatile fatty acids in media containing lactose than in media containing glucose. The preferred metabolism of [14C]substrates, independent of media carbohydrate, was in the following order: lactic acid > galactose, lactose > glucose. The volatile fatty acids in which radiolabel was most concentrated were acetic acid, propionic acid, or butyric acid.

The present investigation describes a novel method of demonstrating strain diversity among Pasteurella haemolytica biovar A, serovar 2 (PhA2) nasal turbinate isolates from a flock of 32 experimental goats during a naturally occurring outbreak of pasteurellosis. After a 21 day conditioning period in a feedyard, 51 PhA2 isolates from 27 culture-positive goats were identified including 1 on day 22, 14 on day 25, 21 on day 39, and 15 on day 66. Each PhA2 isolate was evaluated for its enzyme activity against 19 substrates with a commercial semiquantitative enzyme system and for its antimicrobial susceptibility with 12 drugs, resulting in 7 different enzyme profiles and 8 different antimicrobial susceptibility profiles. A total of 14 combined enzyme and antimicrobial susceptibility profiles were produced. The same PhA2 strain was isolated from only 4 of the 12 goats with 2 PhA2 isolations, while the same PhA2 strain was isolated from only 1 of the 6 goats with 3 PhA2 isolations. The data from this investigation demonstrated that the PhA2 upper respiratory tract flora from goats is highly heterologous.

A defined bacterial culture protective against Salmonella typhimurium cecal colonization in broiler chicks was derived utilizing a continuous-flow (CF) culture apparatus. Chicks receiving the CF culture in combination with a diet containing dietary lactose were protected against cecal colonization by S. typhimurium. The culture consisted of a mixture of gram-positive and gram-negative facultative and strictly anaerobic bacteria. The isolates were identified as Enterococcus avium, two strains of Enterococcus faecalis (designated A and B), Lactococcus lactis, Lactobacillus animalis, a Lactobacillus that could not be identified to species level (designated strain CMS), Citrobacter freundii, Escherichia coli, E. fergusonii, Bifidobacterium animals, and Propionibacterium acidipropionici. Results indicated that CF cultures can be used as a tool to identify bacteria which are antagonistic to S. typhimurium in the chick cecum.

An in vitro continuous-flow (CF) culture system was utilized to isolate and maintain a defined mixed culture of indigenous cecal bacteria from adult broilers. The protective effects of the defined CF culture and dietary lactose on Salmonella typhimurium colonization were evaluated in broiler chicks. The CF culture was administered to chicks by crop gavage on the day of hatch. Lactose was provided as 5% (wt/wt) of the feed ration. The chicks were challenged orally with 10(4) S. typhimurium at 3 days of age and evaluated for Salmonella colonization 7 days after challenge. The experiment was repeated in six separate trials using newly hatched chicks and CF culture that was maintained in continuous steady-state conditions from 42 to 190 days. Compared with controls, the mean number of S. typhimurium in the cecal contents of the chicks given CF culture and dietary lactose decreased significantly (P < .01) by 4.2 log10 units. Similarly, the numbers of Salmonella cecal culture-positive chicks was significantly decreased (P < .01) by 55% in the chicks given CF culture and lactose. The results indicated that a defined culture of indigenous cecal bacteria isolated and maintained in CF culture, together with dietary lactose, effectively controlled S. typhimurium cecal colonization in newly hatched broiler chicks.

The chemotactic and chemotropic responses of the plasmodial stage of the slime mold Physarum polycephalum were used to distinguish Salmonella gallinarum and Salmonella pullorum from 10 Salmonella serovars that are commonly isolated from domestic poultry. Utilizing an in vitro plasmodium agar plate assay method, P. polycephalum was attracted to S. gallinarum and S. pullorum, but the organism was repelled by Salmonella derby, Salmonella dublin, Salmonella enteritidis, Salmonella heidelberg, Salmonella minnesota, Salmonella montevideo, Salmonella newington, Salmonella newport, Salmonella reading, and Salmonella typhimurium.

An epidemiologic study of Pasteurella haemolytica serovar 1 (Ph1) in market-stressed feeder calves from 7 farms in eastern Tennessee was conducted. The nasal mucus of each calf was cultured sequentially at the farm of origin (day 0), at an auction market (day 133), and at a feedyard in Texas (days 141, 148, 155, and 169). Of the 103 calves tested, 77 were culture-positive, including 1 on day 0, 1 on day 133, 20 on day 141, 57 on day 148, 50 on day 155, and 14 on day 169. From the 143 Ph1 isolates, 20 enzyme profiles were determined by use of a commercial enzyme system that detects 19 enzymatic reactions; 4 antimicrobial susceptibility profiles were obtained, using the disk-diffusion method, which evaluated susceptibility to 11 antibacterial drugs. All isolates were positive for acid phosphatase and alkaline phosphatase, but were negative for alpha-galactosidase, alpha-mannosidase, beta-glucosidase, beta-glucuronidase, cystine aminopeptidase, N-acetyl-beta-glucosaminidase, and trypsin. Other positive enzyme reactions included: leucine aminopeptidase, 140 Ph1 isolates; phosphohydrolase, 90 isolates; alpha-fucosidase, 63 isolates; esterase (C4), 59 isolates; valine aminopeptidase, 30 isolates; esterase lipase (C8), 24 isolates; beta-galactosidase, 2 isolates; and alpha-glucosidase, chymotrypsin and lipase (C14), 1 isolate each. Thirty-four Ph1 profiles were identified, using combined enzyme and antimicrobial susceptibility profiles. The data indicate that the strains isolated during the feedyard period may have been determined more by farm of origin (P < or = 0.001) than by habitation with calves from other farms while in the feedyard.(ABSTRACT TRUNCATED AT 250 WORDS)

A Veillonella species and Enterococcus durans were isolated from the cecal contents of adult broilers. Mixed cultures of Veillonella and E. durans inhibited the growth of Salmonella typhimurium and Escherichia coli 0157:H7 on media containing 2.5% lactose (w/v). The growth of S. typhimurium or E. coli 0157:H7 was not inhibited by mixed cultures containing Veillonella and E. durans on media containing only 0.25% lactose or by pure cultures of Veillonella or E. durans on media containing either 0.25% or 2.5% lactose. The mixed cultures of Veillonella and E. durans produced significantly (P<0.05) more acetic, propionic, and lactic acids in media containing 2.5% lactose than in media containing 0.25% lactose. The inhibition of the enteropathogens was related to the production of lactic acid from lactose by the E. durans and the production of acetic and propionic acids from lactic acid by the Veillonella .

The cellular, cultural, and biochemical characteristics of eight isolates of a large gram-positive bacillus that are commonly observed as apparently normal flora in the proventriculus of budgerigars (Melopsittacus undulatus) were determined. The bacterium was highly pleomorphic and changed markedly in both diameter and length when subcultured on agar media. The bacterium was facultative anaerobic and capnophilic, hemolytic on blood agar, and formed flat colonies with irregular edges after incubation for several days. All isolates grew on sodium azide agar but did not grow on MacConkey agar. The isolates were catalase-negative and oxidase-negative and did not reduce nitrate. All isolates failed to utilize arginine, lysine, ornithine or tryptophane but produced acid from glucose, galactose, levulose, maltose, melibiose, starch, and sucrose. All isolates produced acetoin from glucose and hydrolyzed esculin. The eight isolates could not be identified to either genus or species level based on the descriptions of currently classified organisms in the division Firmicutes as described in Bergey's Manual of Systematic Bacteriology.

At an abattoir, lesion specimens from 140 condemned sheep livers were collected for bacteriologic culture and for pathologic examination. Grossly, 23 lesions were abscesses; from 9 of which, Fusobacterium necrophorum biovar A (3 in pure culture and 6 in mixed culture) was isolated and from 14 of which, biovar B (6 in pure culture and 8 in mixed culture) was isolated. Escherichia coli was the predominant facultative anaerobic bacterium and Clostridium perfringens was the predominant obligate anaerobic bacterium isolated from the 14 lesions with mixed bacterial infection. Histologically, these lesions had a core of coagulation necrosis, encircled by a zone of necrotic phagocytic cells and bacteria with cellular characteristics of F necrophorum biovars A or B, and a connective tissue capsule. Of the 117 lesions without F necrophorum, 49 were culture-positive (for other organisms) and 69 were culture-negative. These 117 lesions were fibrous and were smaller than the 23 abscesses. A variety of gram-positive and gram-negative facultative anaerobic and obligate anaerobic bacteria was isolated from the culture-positive lesions, but always in low numbers. Eleven culture-negative and 18 culture-positive lesions were examined and had histologic characteristics of parasite-induced granulomas, with numerous eosinophils and epithelioid giant cells. Results of the study indicated that the histologic appearance of ovine hepatic lesions with F necrophorum was similar to bovine liver abscesses caused by F necrophorum, but unlike bovine liver abscesses, F necrophorum biovar B was isolated more frequently than was biovar A and often in pure culture. Most of the lesions in the condemned livers were parasite-induced granulomas.

Two canine models of clinical anaerobic bacterial infections were developed for the study of the clinical parameters associated with these infections and for evaluation of antimicrobial agents that might be useful in therapy. In model I, a mixed culture of Bacteroides fragilis, B melaninogenicus, and Fusobacterium necrophorum was used as the inoculum. In model II, a mixed culture of B fragilis and Clostridium perfringens combined with an infection enhancer (sterile cinder dust) was used as the inoculum. In both models, reproducible localized pyogenic or gangrenous infections were induced. Clinical signs of fever, depression, and leukocytosis with a left shift were present. A depression in the nonspecific cell-mediated immune response was noticed. The 2 models were used to evaluate clindamycin and lincomycin for therapy of anaerobic bacterial infections, using a subjective scoring system for severity of lesions and general clinical appearance. Clindamycin at dosage levels of 5.5 mg or 11 mg/kg of body weight, twice a day was highly efficacious in the treatment of anaerobic bacterial infections in both models. Dogs given lincomycin (22 mg/kg twice a day) responded to the treatment, but the response was less than that seen with clindamycin.

Ovariectomized cows were treated with either estrogen, progesterone or adrenogorticotropic hormone (ACTH). Each cow's uterus was inoculated with 1x10(6)Corynebacterium pyogenes organisms. The cows were evaluated by rectal examination for signs of uterine infection. Peripheral leukocytic phagocytosis was determined by chemiluminescence at the beginning (Day 4) and end (Day 21) of the experiment. Uterine cultures were obtained at the end. None of the estrogen-treated cows developed signs of uterine infection, but all the other cows did develop uterine disease. All the infected cows showed clinical improvement at the experimental end (Day 21). There were no differences between groups for leukocytic phagocytosis on Day 4, but on Day 21, values for progesterone-treated and control cows were similar but greater than those for estrogen or ACTH-treated cows, which were similar. Leukocytic phagocytosis values for all cows were lower on Day 21 than on Day 4. Most of the estrogen- and ACTH-treated cows had negative intrauterine cultures on Day 21 while most of controls and progesterone-treated cows had positive cultures.

Isolates of Fusobacterium necrophorum were obtained from 124 bovine hepatic abscesses (in 119 cattle) and from the ruminal contents of 12 cattle. Three biotypes (A, AB, and B) were found. Type A isolates were most common in hepatic abscesses, and type B isolates were most common in ruminal contents. Virulence studies in mice indicated that type A isolates were more pathogenic than types AB or B isolates. The difference in virulences between types A and B isolates was not related to the toxicity of the endotoxin. Results of serotesting (passive hemagglutination) showed that a wide heterogeneity existed between isolates. Minimal inhibitory concentrations of antibiotics for 12 of the isolates (7 type A, 1 type AB, and 4 type B) were as follows: less than or equal to 0.06 IU of penicillin G/ml; less than or equal to 0.5 micrograms of cephaloridine/ml; less than or equal to 2 micrograms of chloramphenicol/ml; less than or equal to 0.06 micrograms of clindamycin/ml; less than or equal to 8 micrograms of erythromycin/ml; less than or equal to 128 micrograms of gentamicin/ml; less than or equal to 0.25 micrograms of oxytetracycline/ml; and less than or equal to 4 micrograms of tylosin/ml.

Broilers from commercial flocks experiencing a 10-60% incidence of scabby-hip lesions at processing were examined, and selected skin lesions were cultured. Over 70% of the lesions were associated with traumatic excoriations, particularly on the caudal dorsal convexity of the birds. Most lesions were observed on birds that were 5 weeks of age or older. From the 27 specimens cultured, Clostridium perfringens was isolated in pure culture from 4 lesions and Staphylococcus species from 10 lesions. Pure cultures of staphylococci were recovered from 4 lesions, and 2-5 different staphylococci were isolated from 6 lesions. Eight staphylococci were identified as S. sciuri, 8 as S. simulans, 2 as S. epidermidis, 2 as S. lentus, 2 as S. warneri, 1 as S. cohnii, and 1 as S. intermedius. Fifty cutaneous specimens from 10 5-week-old normal broilers were cultured. A total of 197 isolates were identified including 65 S. sciuri, 52 S. lentus, 24 S. simulans, 12 S. hyicus, 11 S. warneri, 9 S. cohnii, 9 S. gallinarium, 8 S. xylosus, and 7 S. epidermidis.

Calves (n = 150) born to cows infected with field strains of Brucella abortus were weaned within 24 hours after birth or allowed to nurse until weaned (age at weaning ranged from 10 to 43 weeks). After weaning, female calves were placed in small groups in special pens constructed to prevent further direct and environmental exposure to brucellae. At weaning, male calves were moved to an isolated pasture and were kept as a single group. Later, first-generation heifers were bred and placed in individual isolation pens until termination of pregnancy. Two of the first-generation heifers (that were culture-negative) developed persistent serologic responses (to B abortus) of unexplained origin. Brucellae were not isolated from progeny of infected cows (105) or from progeny offspring (95 fetuses and neonates), indicating that latency may be infrequent.

Two studies of brucellosis in wildlife on farms where the brucellosis infection prevalence in cattle was known are reported. On a research farm, 233 feral animals of 22 mammalian species and 12 of seven avian species were trapped during three time periods. Sixty were studied before cattle were introduced, 128 were studied while 501 cattle infected with Brucella abortus were calving and aborting, and 60 specimens were collected 20 mo after the last infected cow calved. Selected tissues from 229 wild animals were cultured and sera from 138 were examined using the brucellosis card, standard tube agglutination (STA), 2-mercaptoethanol (2-ME) and rivanol (RIV) tests. Brucella abortus was not recovered from any animals sampled prior to cattle being introduced and all sera collected were negative. Brucella abortus was isolated from four opossums (Didelphis virginiana) and one raccoon (Procyon lotor) in the group of animals trapped during the calving period. Three serums were tested and had STA titers ranging from 1:100 to 1:200. Of 68 sera only one had antibodies. Brucella were not isolated from 59 animals trapped after the calving period and only one of 42 serums had antibodies. On regional cattle farms, 243 wild animals were trapped. Brucellae were not isolated from 223 animals which were cultured. No serums had significant titers. The data from this study suggest opossums and raccoons can be infected from cattle but are unlikely to maintain the infection.

The in vitro leukotoxic activity of 3 bovine isolates of Fusobacterium necrophorum which varied in pathogenicity were compared. Monolayers of mouse peritoneal macrophages were exposed to culture filtrates from each F necrophorum strain, and cell viability was determined, using the trypan blue dye exclusion test. Two methods were used for production of the leukotoxin: (1) medium M-1 continuous dialysis sac cultures and (2) brain-heart infusion agar plate cultures. Supernatant cultural fluids containing the leukotoxin were subjected to membrane-partition chromatography, using ultrafilters with approximate molecular weight (mol wt) exclusion limits of 100,000, 10,000, 2,000, and 500. All ultrafiltrates had a cytotoxic effect on the monolayers. Cytotoxic activity was not found in the ultrafilter residues or in the control media ultrafiltrates. Comparative study of leukotoxin production indicated that F necrophorum 2101, type A, produced the most leukotoxin; F necrophorum 2030, type AB, produced slightly less leukotoxin; and F necrophorum 2035, type B, produced small amounts of leukotoxin. Endotoxin activity, as demonstrated by the mouse lethality test, was found in the residues of the XM-100A ultrafilter (100,000 mol wt), but not in the filtrates. Culture supernatant fluids and the XM-100A ultrafiltrates were positive for endotoxin, using the limulus amebocyte lysate assay; however, the other ultrafiltrates with lower mol wt exclusion limits were negative.

Two studies of experimentally induced Brucella abortus infections in dogs are reported. Twenty dogs in experiment 1 were fed approximately 4.8 X 10(10) colony forming units of B. abortus strain 2308 (BA 2308), and 11 dogs in experiment 2 were fed approximately 3.4 X 10(14) BA 2308. Serum samples from each infected dog and noninfected control were tested on the day of infection and at weekly and biweekly intervals post-infection (PI) for antibodies to B. abortus. The standard tube agglutination (STA) and rivanol (RIV) mean log-titers of the infected dogs in the 2 experiments were compared statistically on the day of infection and on PI days 7, 14, 21, 35, 42 and 49. The STA and RIV titers of the infected dogs in experiment 2 were significantly higher than were those of the dogs in experiment 1. Brucella abortus was isolated from 29 infected dogs in experiments 1 and 2 including 10 dogs, which were seronegative when cultured.

Forty patients presenting for vaginal termination of pregnancy, divided randomly into four groups, received either no medication, sodium citrate 30 ml orally, ranitidine 150 mg orally or ranitidine 50 mg intravenously. During the procedure, gastric contents were removed by orogastric tube for volume and pH measurements. Ranitidine, orally and intravenously, significantly increased gastric pH and reduced gastric volume. In the control group only one pH was greater than 2.5. Sodium citrate raised the pH above 2.5 in 6 out of 10 patients. Fasting patients in the first months of pregnancy may be at risk of developing Mendelson's syndrome. Ranitidine is very effective in increasing gastric pH and at the same time reducing gastric volume in such patients.

The biochemical characteristics of the leukotoxins of 3 bovine isolates of Fusobacterium necrophorum which represent biotypes A, AB, and B were compared. Two methods were used for the production of the leukotoxins: medium M-1 continuous dialysis sac cultures and brain-heart infusion agar plate cultures. The supernatant cultural fluids were fractionated sequentially by membrane-partition chromatography, using ultrafilters with approximate molecular weight (mol wt) exclusion limits of 100,000, 10,000, 2,000, and 500. The ultrafiltrates (less than 500 mol wt) were fractionated by gel-permeation chromatography, using G-10 Sephadex. The leukotoxins of the 3 F necrophorum strains were estimated to have a molecular weight between 350 and 450. The leukotoxins in the ultrafiltrates (less than 500 mol wt) were stable at 60 C for 4 hours and at 100 C for 30 minutes, stable to extremes of pH (3 to 11), and stable to degradative enzymes including trypsin, protease, alpha-amylase, lipase, deoxyribonuclease, and ribonuclease. Significant differences were not observed in the biochemical characteristics of the leukotoxins produced in vitro by the 3 F necrophorum biotypes. These assays were done, using monolayers of mouse peritoneal macrophages. The monolayers were exposed to the 4 ultrafiltrates of both the continuous dialysis sac and brain-heart infusion agar cultures (pH 7.2) for 4 hours at 4 C, 25 C, and 37 C. Maximal cytotoxic activity in the assays was at 37 C.

Ten gray foxes, eight principals that were fed approximately 4.4 X 10(10) colony forming units of Brucella abortus strain 2308 and two controls, were examined for serologic responses and tissue distribution of the organisms. Blood sera from each fox were tested on the day of exposure and at seven weekly intervals for antibodies to B. abortus, using the brucellosis card, standard tube agglutination, 2-mercaptoethanol and rivanol tests. Control foxes were serologically negative for all tests throughout the study and the principals were negative prior to exposure. On days 14, 21 and 28, the eight principals had positive card reactions and greater than or equal to 1:100 tube agglutination titers. After 28 days, the titers receded; and by day 49, three principals had negative card reactions and one of these was negative for all tests. Brucella abortus was isolated from one or more lymph nodes from seven of eight principals including the one which was seronegative. The bacterium was not isolated from lungs, livers, spleens, kidneys, uteri or testicles.

Cellular, colonial, cultural, and biochemical characteristics of 25 field strains of gram-negative pleomorphic bacilli from rams with epididymitis were compared with Actinobacillus actinomycetemcomitans American Type Culture Collection (ATCC) strain 29522 and Actinobacillus seminis ATCC strain 15768. Three field strains were identified as A. actinomycetemcomitans, 15 as A. seminis, and 2 as Haemophilus agni; however, 5 strains (3 in group A and 2 in group B) were not identified as species in the genera Actinobacillus, Haemophilus, or Pasteurella based on the taxonomic criteria in Bergey's manual of systematic bacteriology. The 5 Actinobacillus-like organisms in groups A and B were predominantly gram-negative coccobacilli and exhibited less pleomorphism than the 2 Actinobacillus species. The colonial morphologies of groups A and B were similar to the 2 Actinobacillus species but were smaller in diameter and had a pale yellow color. Groups A and B, like the actinobacilli, were facultative anaerobic and capnophilic, did not grow on MacConkey agar, and were catalase-positive and oxidase-positive. Group A reduced nitrate but group B did not. The A. seminis strains utilized ornithine, and group A utilized arginine; but group B did not utilize either ornithine or arginine. All strains failed to utilize lysine or tryptophane. All strains produced acid but no gas from glucose, and the utilization of other carbohydrates varied markedly both between and within the 5 groups of bacteria.(ABSTRACT TRUNCATED AT 250 WORDS)

Fusobacterium necrophorum is considered to be a member of the normal rumen flora and is the primary etiologic agent of bovine liver abscesses. Of the three biotypes of F. necrophorum, A, B, and C, only biotypes A and B have been implicated in the disease. Type B is the predominant biotype isolated from ruminal lesions and type A is the predominate biotype isolated from liver abscesses. Type A is usually found in pure culture in the liver abscesses; whereas, type B is usually found in mixed culture with either type A or with other bacterial species. Corynebacterium pyogenes, Streptococcus spp., Staphylococcus spp., and Bacteroides spp. are the most prevalent bacteria recovered from mixed cultures. Corynebacterium pyogenes is the most common species isolated and can cause disease synergistically with type B isolates.

A survey for anaerobic bacteria was conducted in 314 clinical specimens from dogs and cats. A total of 187 anaerobic isolates in pure and mixed culture were isolated from 111 of the specimens that contained anaerobic bacteria. Common isolated included Actinomyces (9.1%), Clostridium perfringens (19.3%), other Clostridium spp (11.2%), Peptostreptococcus anaerobius (7.5%), Bacteroides melaninogenicus (13.4%), other Bacteroides spp (17.6%), and Fusobacterium necrophorum (5.3%). Anaerobic bacteria were involved in serious lesions that often were life threatening to the animals. Antibiotic susceptibility data indicated that the lincomycin family, the penicillin family, chloramphenicol, and cephaloridine are preferred drugs for treatment of anaerobic infections. Data from the survey were used in formulation of a table to aid practitioners in clinical diagnosis of disease caused by anaerobes. Clostridium perfringens was isolated in large numbers from five of six dogs with a clinical diagnosis of canine hemorrhagic gastroenteritis and from one cat with hemorrhagic diarrhea. Experimental infections were induced in rats, using caine feces as inoculum. Induced lesions contained aerobic and anaerobic bacteria similar to those bacteria isolated in the clinical survey, indicating that feces may serve as a major source of these bacteria in clinical infections of the dog.

Viable cultures of Fusobacterium necrophorum were inoculated into the hepatic portal veins of 7 cattle. The experimentally produced hepatic necrobacillosis lesions were described from 30 minutes through 36 hours. The lesions were established by 2 different host-bacterial interactions. Most frequent, microabscesses were initiated by diffusely distributed bacteria and were observed from 30 minutes through 2 hours. Gross abscesses from 3 through 8 hours had centrally located abscess cores and were encircled by leukocytes which were predominantly neutrophils. Lesions from 12 through 36 hours had abscess cores that were encircled by leukocytes which were predominantly macrophages. Less frequent, emboli of F. necrophorum initiated infections in hepatic sinusoids or telangiectatic lesions. Hepatocytes adjacent to the bacteria exhibited coagulation necrosis and few leukocytes were observed in these lesions.

Thirteen female dogs, which included eight principals that were fed approximately 4.4 X 10(10) colony forming units (cfu) of Brucella abortus strain 2308 and five sentinels that were housed with the principals, were examined for serologic responses, blood culture, tissue distribution of the organisms and pathologic lesions. Serum samples from each dog were tested on the day of exposure and on post exposure days 5, 7, 10, 14, 21, 28, 35, 42 and 49 for antibodies to B. abortus, using the brucellosis card (BC), standard tube agglutination (STA), 2-mercaptoethanol (ME) and rivanol (RIV) tests. Antibodies were detected in the principals by day 5 and increased through day 21. The STA test was the first to become positive, followed by the BC, ME and RIV tests. After 28 days, the serologic titers receded. From day 14 through day 42, all principals had greater than or equal to 1:50 STA titers. On day 49, seven principals had greater than or equal to 1:50 STA titers and one had a 1:25 STA titer. The sentinels were negative for all tests, except sentinel number 9 which had STA titers ranging from 1:25 to 1:50 on day 14 through day 35. Blood cultures that were obtained from each principal at intervals from one hour after exposure through 49 days were negative. Brucella abortus was isolated from various lymph nodes of the eight principals and from sentinel number 9, which was apparently infected by ingesting brucellae contaminated feces from the principals. Microscopic lesions were not observed in the culture-positive tissues examined.