Impact of an experimental PRRSV and Streptococcus suis coinfection on the pharmacokinetics of ceftiofur hydrochloride after intramuscular injection in pigs.

This study determined the impact of porcine reproductive and respiratory syndrome virus (PRRSV) and Streptococcus suis coinfection on the pharmacokinetic (PK) profile of ceftiofur hydrochloride in pigs after intramuscular (i.m.) injection. Eighteen clinically normal crossbred gilts were assigned by weight into a challenge group (10 pigs) and control group (eight pigs). Pigs in both groups received a single i.m. injection of ceftiofur hydrochloride (Excenel RTU Sterile Suspension; Zoetis) at a 5 mg/kg BW dose. Serial blood samples were collected to characterize the plasma concentration curve. After a 10 days drug washout period, the challenge group was inoculated with 2 mL of PRRSV isolate VR-2385 (10(5.75) 50% tissue culture infective doses per mL) intranasally and 8 days later inoculated S. suis. When clinical disease was evident, the second PK assessment began in both challenge and control groups. Coinfected pigs demonstrated lower values of AUC and CMAX , but higher values of Cl/F and Vz/F indicating drug kinetics were altered by infection. The data from this study have implications on ceftiofur treatment regimens in diseased pigs.

Comparison of sesion severity, distribution, and colonic mucin expression in pigs with acute swine dysentery following oral inoculation with "Brachyspira hampsonii" or Brachyspira hyodysenteriae.

Swine dysentery is classically associated with infection by Brachyspira hyodysenteriae, the only current officially recognized Brachyspira sp. that consistently imparts strong beta-hemolysis on blood agar. Recently, several strongly beta-hemolytic Brachyspira have been isolated from swine with clinical dysentery that are not identified as B. hyodysenteriae by PCR including the recently proposed species "Brachyspira hampsonii." In this study, 6-week-old pigs were inoculated with either a clinical isolate of "B. hampsonii" (EB107; n = 10) clade II or a classic strain of B. hyodysenteriae (B204; n = 10) to compare gross and microscopic lesions and alterations in colonic mucin expression in pigs with clinical disease versus controls (n = 6). Gross lesions were similar between infected groups. No histologic difference was observed between infected groups with regard to neutrophilic inflammation, colonic crypt depth, mucosal ulceration, or hemorrhage. Histochemical and immunohistochemical evaluation of the apex of the spiral colon revealed decreased expression of sulphated mucins, decreased expression of MUC4, and increased expression of MUC5AC in diseased pigs compared to controls. No difference was observed between diseased pigs in inoculated groups. This study reveals significant alterations in colonic mucin expression in pigs with acute swine dysentery and further reveals that these and other microscopic changes are similar following infection with "B. hampsonii" clade II or B. hyodysenteriae.

Influence of ampicillin, ceftiofur, attenuated live PRRSV vaccine, and reduced dose Streptococcus suis exposure on disease associated with PRRSV and S. suis coinfection.

The objective of this research was to evaluate the efficacy of two antimicrobials (ampicillin and ceftiofur), a modified-live porcine reproductive and respiratory syndrome virus (PRRSV) vaccine, and low dose exposure to Streptococcus suis on disease associated with PRRSV/S. suis coinfection. Fifty-six, crossbred, PRRSV-free pigs were weaned at 10-12 days of age and randomly assigned to five treatment groups. All pigs were inoculated with 2ml of 10(6.4) TCID50/ml of high virulence PRRSV isolate VR-2385 intranasally at 29-31 days of age (day 0 of the study) followed 7 days later by intranasal inoculation with 2ml of 10(8.9)colony forming units(CFU)/ml S. suis type 2 isolate ISU VDL #40634/94. Pigs in group 1 (n=10) served as untreated infected positive controls. Pigs in group 2 (n=12) were treated with 5.0 mg/kg ceftiofur hydrochloride intramuscularly (IM) on days 8, 11, and 14. Pigs in group 3 (n=11) were treated with 11 mg/kg ampicillin IM on days 8-10. Pigs in group 4 (n=12) were vaccinated 14 days prior to PRRSV challenge with a commercial modified-live PRRSV vaccine. Pigs in group 5 (n=11) were exposed to a 1:100 dilution of the S. suis challenge inoculum 19 days prior to S. suis challenge. Mortality was 80, 25, 82, 83, and 36% in groups 1-5, respectively. The reduced dose S. suis exposure had some residual virulence, evidenced by S. suis induced meningitis in two pigs after exposure. Treatment with ceftiofur hydrochloride and reduced dose exposure to S. suis were the only treatments which significantly (P<0.05) reduced mortality associated with PRRSV/S. suis coinfection, significantly (P<0.05) reduced recovery of S. suis from tissues at necropsy, and significantly (P<0.05) reduced the severity of gross lung lesions.

In vitro activity of four antimicrobial agents against North American isolates of porcine Serpulina pilosicoli.

Porcine colonic spirochetosis is a nonfatal diarrheal disease that affects pigs during the growing and finishing stages of production. The disease is caused by Serpulina pilosicoli, a newly recognized species of pathogenic intestinal spirochete. Antimicrobial therapy aimed at reducing the infection may be helpful in controlling spirochetal diarrhea. In this study, the in vitro antimicrobial susceptibilities of the reference isolate S. pilosicoli P43/6/78 from the United Kingdom and 19 field isolates obtained from pigs in Canada (n = 5) and the United States (n = 14) were determined against the antimicrobial agents carbadox, gentamicin, lincomycin, and tiamulin, all of which are commonly used for control of the related pathogenic intestinal spirochete S. hyodysenteriae. Additionally, the susceptibility or resistance of each isolate against each antimicrobial agent was estimated on the basis of available data on the in vitro antimicrobial susceptibility breakpoints of S. hyodysenteriae. Each isolate was identified on the basis of phenotypic and genotypic markers, and the minimum inhibitory concentration of each antimicrobial agent was determined by the agar-dilution method. All the isolates were susceptible to carbadox and tiamulin. The percentages of isolates susceptible, intermediate, and resistant to lincomycin were 42.1%, 42.1%, and 15.8%, respectively. Slightly less than half of the isolates (47.4%) were susceptible to gentamicin, and the remainder (52.6%) were resistant. Implementation of rational control measures to reduce infection by S. pilosicoli should improve overall health and productivity in swine herds.

Bacterial isolates from plaque and from blood during and after routine dental procedures in dogs.

OBJECTIVE: This study evaluates the association between dental procedures and bacteremia in dogs, including a comparison of bacteria isolated from plaque and blood, severity of the bacteremia versus the severity of dental disease, and the longevity of bacteremia. STUDY DESIGN: Bacteria cultured from the blood over time were compared with those isolated from the plaque and crevicular fluid and in relation to severity of dental disease. ANIMALS OR SAMPLE POPULATION: Twenty adult greyhounds. METHODS: Blood samples were collected for culture before induction of general anesthesia, immediately after intubation, 20 minutes after initiation of the dental procedure, and at 10-minute intervals until 10 minutes after the dental procedure was completed. Samples of plaque were taken for microbiological culture. RESULTS: Sixty to ninety percent of the bacterial genera isolated from the plaque were present in the blood. Dogs classified according to severity of dental disease showed no difference in the total number of different species or number of different Gram-negative, Gram-positive, or anaerobic bacteria isolated from plaque or blood (P < .05). Bacteremia was present in all of the dogs studied, within 40 minutes from the initiation of the dental procedure, regardless of the severity of oral disease. CONCLUSIONS: Gram-negative, Gram-positive, and anaerobic bacteria are present in blood during dental procedures; the bacteremia can persist beyond the dental procedure, and is not associated with the severity of dental disease. CLINICAL RELEVANCE: The nature and extent of bacteremia occurring during routine dental procedures is important in understanding a potential risk to dogs.

Morphometric evaluation of immunoglobulin A-containing and immunoglobulin G-containing cells and T cells in duodenal mucosa from healthy dogs and from dogs with inflammatory bowel disease or nonspecific gastroenteritis.

OBJECTIVE: To investigate the distribution of IgA- and IgG-containing cells and T cells in the villi of duodenal mucosa from healthy dogs and from dogs with inflammatory bowel disease (IBD) of gastroenteritis. DESIGN: Case-control study. ANIMALS: 28 dogs, grouped according to clinical and histologic criteria: 11 dogs with IBD, 8 dogs with non-specific gastroenteritis, and 9 healthy dogs. PROCEDURE: Endoscopic biopsy specimens of duodenal mucosa from each dog were stained specifically for IgA and IgG heavy chains and pan T-cell (CD3) antigen, using immunoperoxidase techniques. Morphometric analysis, performed via an image-analysis system, was used to count IgA- and IgG-containing cells and T cells within paired contiguous villi from each dog. RESULTS: cells were the predominant immune cell type in all groups of dogs. Significant differences in the villus distribution of IgA- and IgG-containing cells and T cells were not observed. Healthy dogs had significantly higher T-cell counts than had dogs with IBD or gastroenteritis. Dogs with nonspecific gastroenteritis had a significantly higher concentration of IgA-containing cells than the other groups of dogs had. Significant group differences for IgG-containing cells also were evident, with dogs with IBD having the lowest cell counts. CONCLUSIONS AND CLINICAL RELEVANCE: High concentrations of IgA- and IgG-containing cells and T cells in the villus lamina propria cannot be reliably used to distinguish IBD from other intestinal disorders in dogs. Evaluation of T cells may be the most discriminatory method for differentiating dogs with IBD from clinically normal dogs via examination of intestinal biopsy specimens.

Bacterial and mycoplasmal flora of the healthy camelid conjunctival sac.

Healthy conjunctival sacs of 88 animals of 3 species of captive camelids (Lama glama, Lama guanicoe, Lama pacos) and llama-guanaco hybrids were sampled for bacterial and mycoplasmal flora. Mycoplasmas were not isolated from any animal. Eleven genera of bacteria were isolated. The most frequent isolates were Staphylococcus epidermidis and Pseudomonas spp. Nine varieties of Pseudomonas were found, which represented at least 3 Pseudomonas species. Many of the bacterial isolates (especially the pseudomonads) are potential pathogens in the eyes of these camelids.

Improved selective medium for the isolation of Treponema hyodysenteriae.

An agar medium with improved selection for Treponema hyodysenteriae was developed. Cultures of T. hyodysenteriae and T. innocens, feces from 11 clinically normal pigs, and colonic contents from 6 pigs with gross lesions consistent with swine dysentery were diluted in phosphate-buffered saline and plated on Trypticase soy agar (BBL Microbiology Systems, Cockeysville, Md.) with 5% citrated bovine blood (TSA), TSA with 400 micrograms of spectinomycin per ml (TSA-S400), TSA-S400 with 25 micrograms each of colistin and vancomycin per ml, and TSA with 5% pig feces extract and five antimicrobial agents (spiramycin, rifampin, vancomycin, colistin, and spectinomycin) (BJ). Viable numbers of T. hydodysenteriae grown on BJ were virtually identical to those for TSA, TSA-S400, and TSA-S400 with colistin and vancomycin. Pure cultures of four isolates of T. hyodysenteriae and three isolates of T. innocens were sustained through six subcultures on BJ. Fecal floras were completely inhibited on BJ for 14 of 17 fecal samples from both groups of pigs. A total of 461 colonic specimens from naturally occurring cases of porcine intestinal disease were plated on TSA-S400 and BJ. T. hyodysenteriae was isolated on both TSA-S400 and BJ for 69 specimens and on BJ alone for an additional 19 specimens.

Autoclaved liquid medium for propagation of Treponema hyodysenteriae.

Three liquid media that differ slightly in composition but not in the method of preparation were developed for the propagation of Treponema hyodysenteriae and Treponema innocens. The three media are unique in that all components are sterilized by autoclaving before use. These media supported better growth of T. hyodysenteriae than did previously used liquid media.

Isolation of Treponema hyodysenteriae from wild rodents.

Rodents from swine-producing farms were examined for the presence of Treponema hyodysenteriae. Wild mice (n = 257) and rats (n = 41) were trapped on eight farms. Ceca were removed aseptically, and the contents and mucosal scrapings were cultured on selective medium (blood agar containing 400 micrograms of spectinomycin per ml). T. hyodysenteriae was detected in the cecal scrapings of four mice from three different farms where swine dysentery had occurred. Gross lesions were detected in the ceca in two of the four mice. In addition, Treponema innocens was detected in the cecal scrapings of 12 mice and 13 rats. Three of the four T. hyodysenteriae isolates were pathogenic when inoculated intragastrically into swine. The results of this investigation suggest that wild rodents may be carriers of T. hyodysenteriae.

Enzyme-linked immunosorbent assay for detection of antibody to Treponema hyodysenteriae antigens.

The enzyme-linked immunosorbent assay (ELISA) was evaluated and compared with the microtitration agglutination test for the detection of swine antibody to Treponema hyodysenteriae lipopolysaccharide antigens. Cells of T. hyodysenteriae serotypes 1 and 2 were extracted with hot phenol-water (68 degrees C). The lipopolysaccharide fraction from the aqueous phase was coated on plastic wells at concentrations of 1 micrograms (serotype 1) and 10 micrograms (serotype 2) of carbohydrate per ml. The ELISA was serotype specific when lipopolysaccharide antigens were reacted against sera from convalescent swine. Seroconversion of infected pigs was detectable with the ELISA within 1 to 2 weeks postinoculation and with the microtitration agglutination test 2 to 3 weeks postinoculation. Antibody titers could be detected in convalescent pigs as long as 19 weeks postinoculation by the ELISA and 12 to 13 weeks postinoculation by the microtitration agglutination test. Therefore, the ELISA may be useful for the detection of asymptomatic carriers.

Differentiation of Treponema hyodysenteriae from T innocens by enteropathogenicity testing in the CF1 mouse.

Fourteen isolates of Treponema hyodysenteriae and 11 isolates of T innocens from eight different countries were evaluated in the CF1 strain female mouse for virulence. Mice were fasted for 24 hours and inoculated intragastrically with 1 ml of culture for two consecutive days. Mice were killed and necropsied at 12 to 15 days after inoculation. Caecitis was detected in mice from each of the groups receiving T hyodysenteriae (67 per cent) but not in mice receiving T innocens or sterile broth. Lesions consisted of hyperaemia, mucosal oedema, catarrhal inflammation and occasional haemorrhage. These studies suggest that the CF1 mouse may be an inexpensive in vivo means of differentiating T hyodysenteriae from T innocens.

Comparison of selective culture and serologic agglutination of Treponema hyodysenteriae for diagnosis of swine dysentery.

Samples of faeces and of serum were collected from pigs of various ages on 21 farms. Faecal samples were cultured on trypticase soy agar containing 5 per cent citrated bovine blood and 400 micrograms per ml spectinomycin, incubated at 42 degrees C in Gaspak jars under an atmosphere of 80 per cent hydrogen: 20 per cent carbon dioxide. Antibody titres to Treponema hyodysenteriae were determined by a microtitration agglutination method using merthiolate-inactivated whole cell antigen prepared from a beta-haemolytic isolate. Results indicated that mean titres in pigs from which beta-haemolytic T hyodysenteriae was isolated were significantly higher than in pigs which yielded isolates of weak beta-haemolytic T innocens or in culturally negative pigs (P less than 0.0225). Mean titres of herds where beta-haemolytic T hyodysenteriae was isolated were significantly higher (P less than 0.005) than the mean titres of either of the other two groups. However, mean titres of herds where no isolates were obtained were not significantly different from mean titres of herds where weak beta-haemolytic T innocens was isolated.

Microtitration agglutination for detection of Treponema hyodysenteriae antibody.

A microtitration agglutination test for the detection of Treponema hyodysenteriae antibody in swine and rabbit sera is described. The following methods provided the best test results: antigen produced from the spirochete after a culturing period of 36 to 44 h at 38 degrees C, washed antigen inactivated with 0.01% Merthiolate at 4 degrees C for 24 to 36 h, sera heated at 56 degrees C for 30 min, a diluent of phosphate-buffered saline (0.01 M, pH 7.2), and test results read macroscopically after 18 to 24 h of incubation at 38 degrees C. The test enabled detection of antibody against pathogenic T. hyodysenteriae with a high level of consistency and sensitivity. Sera against nonpathogenic T. hyodysenteriae produced low agglutinating titers (less than or equal to 1:8) when reacted against antigen from pathogenic isolates. Inactivated antigen remained stable for 7 to 10 days. Specificity of the reaction in the agglutination test was shown by absorption studies.

Enteropathogenicity testing of Treponema hyodysenteriae in ligated colonic loops of swine.

Multiple, ligated loops of swine colon were used as an in vivo model in which to test enteropathogenicity of isolates of Treponema hyodysenteriae. Gross and microscopic lesions observed in 21 of 22 colonic loops in pigs killed 48 to 72 hours after inoculation with isolates known to be enteropathogenic were characteristic of swine dysentery. These lesions were not observed in 12 loops exposed to uninoculated media or in 12 loops inoculated with nonpathogenic isolates of T hyodysenteriae. The swine-loop technique provides a relatively rapid, economical, reliable model in which to test enteropathogenicity of T hyodysenteriae isolates.

Enteropathogenicity of various isolates of Treponema hyodysenteriae.

Isolates of Treponema hyodysenteriae from 25 geographically separated outbreaks of swine dysentery were tested for their ability to produce the disease. Clinical signs and lesions typical of acute swine dysentery were produced in 52 of 68 (75%) susceptible specific pathogen-free pigs that had been orally inoculated with pure cultures of 23 of 25 beta-hemolytic isolates. In addition, 13 weakly beta-hemolytic isolates of nondysentery origin with morphology similar to T. hyodysenteriae did not produce disease when orally inoculated into susceptible specific pathogen-free pigs. Two of these latter isolates, Puppy and B296, and one pathogenic, beta-hemolytic isolate failed to produce disease when orally inoculated into puppies.