Tonsillar Swabs Research Articles

Detection of extracellular factor-positive Streptococcus suis serotype 2 strains in tonsillar swabs of live sows by PCR

In this study, the sensitivity (Se) and specificity (Sp) of a PCR for the detection of EF-positive Streptococcus suis serotype 2 strains in tonsillar swabs of live sows were assessed. We sampled 471 sows originating from four farrow-to-finish farms by tonsillar swabbing and collected their tonsils after slaughter. On these specimens, a PCR, a bacterial examination (BE) or both were performed for the detection of EF-positive S. suis serotype 2 strains. Swab-PCR, Tonsil-PCR and Tonsil-BE were regarded as three integral tests. A Bayesian approach was used to calculate the Se and Sp of the tests. Se and Sp for Swab-PCR were 0.63 (95% credibility interval <0.52, 0.74>) and 0.96 (<0.92, 0.99>), respectively. Values for Se and Sp of Tonsil-PCR amounted to 0.88 (<0.75, 0.96>) and 0.94 (<0.87, 0.99>), respectively. For Tonsil-BE, Se was 0.65 (<0.51, 0.76>) and Sp was 0.97 (<0.91, 0.99>). Repetition of the swabbing procedure after 10 min resulted in a higher Se 0.85 (<0.67, 0.96>) than the Se of the first-Swab-PCR. Repetition of the PCR on the same sample did not result in any significant changes in the outcome of the analysis.

Investigation into the causes of canine infectious respiratory disease: antibody responses to canine respiratory coronavirus and canine herpesvirus in two kennelled dog populations

Summary.Two training centres for working dogs were monitored for one year to determine the presence of viruses and viral antibodies and their association with canine infectious respiratory disease (CIRD). Tonsillar swabs and serum were obtained from dogs on entry into the kennels and in regular intervals thereafter. Additional samples were collected during outbreaks of CIRD. The swabs were examined by virus culture and PCR for canine parainfluenza virus, canine adenovirus, canine herpesvirus (CHV) and canine respiratory coronavirus (CRCoV). Furthermore the prevalence of antibodies to CHV and CRCoV was determined. During this study CIRD was reported mainly in one of the two kennels investigated. In that kennel antibody responses to CRCoV indicated a seasonal occurrence of the virus, which coincided with two outbreaks of respiratory disease. CHV antibody responses were detected throughout the year. In the other kennel, which reported few cases of CIRD a high prevalence of antibodies to CRCoV was detected on entry but only sporadic seroconversions to CRCoV or CHV. By PCR three dogs were found positive for CRCoV in one kennel whereas all PCR tests for other viruses were negative for both kennels. Virus culture failed to detect any viruses in either kennel.

Saliva and meningococcal transmission.

1Neisseria meningitidis carriage was compared in swab specimens of nasopharynx, tonsils, and saliva taken from 258 students. We found a higher yield in nasopharyngeal than in tonsillar swabs (32% vs. 19%, p<0.001). Low prevalence of carriage in saliva swabs (one swab [0.4%]) suggests that low levels of salivary contact are unlikely to transmit meningococci. I nvasive meningococcal disease has a high case-fatality rate and an immediate risk of further cases among household contacts. Public health measures therefore include prompt identification of contacts for chemoprophylaxis (1). One question that commonly arises is whether salivary contact through sharing cups or glasses is an indication for prophylaxis, but the evidence base to inform an answer is weak, and national guidelines are inconsistent (1,2). Although saliva is thought to inhibit meningococcal growth (3), carriage rates in saliva are not known, and swabs to detect carriage are usually taken from tonsils or nasopharynx (4–6). We compared meningococcal isolation rates in swabs of saliva (front of mouth), tonsils, and nasopharynx. We recruited volunteers among students from two colleges in Hereford, England. After giving written consent, students completed a short questionnaire on age, sex, smoking, recent antimicrobial drug use, and meningococcal vaccine status. Three sterile, dry, cotton-tipped swabs were used to take samples from each volunteer: one from the nasopharynx (through the mouth and swept up behind the uvula), one from both tonsils, and one swab of saliva between the lower gum and lips. Swabs were plated directly onto a selective culture medium primarily designed for the isolation of pathogenic Neisseria species (modified New York City base containing vancomycin, colistin, and trimethoprim), prepared by Taunton Media Services, UK (7). The plates were transported to Hereford Public Health Laboratory, where they were spread once from the primary inoculum and incubated in 7% CO 2 at 37°C for 48 h. Putative Neisseria species isolated were sent to the Meningococcal Reference Unit, Manchester Public Health Laboratory, for Neisseria meningitidis confirmation and serologic phenotypic characterization. Data were entered into the computer using Excel (Microsoft Corp., Redmond, WA). Carriage rates by site were compared with McNemar’s test and by risk factor using chi-square tests. Ethical approval was obtained from the Public Health Laboratory Service Ethics Committee and Herefordshire District Ethics Committee. Of the 258 participants, 90 (35%) were identified as carrying Neisseria meningitidis from one or more sites. The site with the highest yield was the nasopharynx (32.2%), whereas tonsillar carriage was 19.4% (Table). One (0.4%) of the 258 saliva swab specimens was positive. No one had positive specimens from all three sites, and the person with the positive saliva swab had negative swabs from the other two sites. Differences in carriage rates between the nasopharynx and tonsils and between the nasopharynx and saliva were statistically significant (p<0.001 in both cases). The predominant serogroup among carried strains was B. No serogroup C strains were identified. Of the 44 carriers with positive swabs from both nasopharynx and tonsils, each pair of isolates was considered to be phenotypically indistinguishable by serogroup, serotype, and sero-subtype. In three of these pairs, one isolate expressed serogroup B, and the paired isolate could not be serogrouped but had identical serotype and sero-subtype. Of the 258 participants, 116 (45%) were men, and 142 were women. Most (86%) were 18 to 21 years of age. Carriage rates were higher among men than women (54/116 vs. 36/142, p<0.001), and among smokers than nonsmokers (49/90 vs. 51/168, p<0.001). Carriage rates were similar when persons were stratified by age, meningococcal vaccination status, and recent antimicrobial drug use. Although duplicate swabs from the nasopharynx sometimes yield different meningococcal strains (3), none of the paired isolates in this study were distinguishable by phenotype. The yield of meningococci from nasopharyngeal swabs was nearly twice as high as that from tonsillar swabs. Previous researchers have found a lower sensitivity of nasopharyngeal swabs taken through the nose using small cotton-tipped wire swabs compared to tonsillar swabs taken using larger cotton-tipped swabs (5,6). Our use of the same type of swab to sample both sites provided a more valid comparison. The carriage rate was higher than expected for this age group (4), suggesting that we had

Presence of macrolide resistance genes in streptococci and enterococci isolated from pigs and pork carcasses

Macrolide and lincosamide (ML)-resistant streptococci and enterococci from tonsillar and colon swabs from 33 pigs and 99 pork carcasses swabs from animals originating from different farms in Belgium were isolated, and their ML resistance phenotypes and genotypes were determined by disk diffusion test and PCR assay, amplifying the ermB gene and the mefA gene. From each of the 33 pigs and 88 of the 99 carcasses' swabs, at least one resistant strain was isolated. The predominant phenotype was the constitutively expressed macrolides, lincosamides and streptogramin B (MLS B) phenotype. This phenotype was most often encoded by the ermB gene. A minority of the strains showed the M phenotype encoded by the mefA gene in streptococci, or the L or ML phenotype.

Actinobacillus pleuropneumoniae infections in closed swine herds: infection patterns and serological profiles

Many farrow-to-finish herds are endemically infected with Actinobacillus pleuropneumoniae. In order to control the disease efficiently, a better knowledge of the ages at which pigs become infected is necessary. Furthermore, no information is available concerning the influence of maternally derived antibodies on the colonization of the upper respiratory tract. Therefore, A. pleuropneumoniae infection patterns were studied in five farrow-to-finish pig herds (A–E) with a history of pleuropneumonia. A longitudinal study was carried out in herds A and B. In these herds, piglets from sows carrying A. pleuropneumoniae in their noses or tonsils were sampled. Nasal and tonsillar swabs as well as sera, were collected from these animals at the age of 4, 8, 12, 16 (herds A and B) and 23 weeks (herd B). At these ages other pigs from the same sows were euthanized. The lungs were macroscopically examined and samples from nose, tonsils and lungs were collected at necropsy. A cross-sectional study was performed in herds C–E. In these herds nasal and tonsillar swabs, as well as sera, were taken from 10 animals of 4, 8, 12 and 16 weeks of age. Lung, nasal and tonsillar samples were tested for the presence of A. pleuropneumoniae by routine bacteriology and PCR with mixed bacterial cultures. The sera were examined for the presence of Apx toxin neutralizing antibodies. In herd A, A. pleuropneumoniae serotype 2 and 10 strains were isolated, whereas serotype 2, 3, 5b and 8 strains were demonstrated in herd B. In most herds, A. pleuropneumoniae was detected in mixed bacterial cultures of tonsillar and/or nasal samples by PCR from the age of 4 weeks onwards. Colonization of the lungs and development of lung lesions was observed in 12- and 16-week-old animals of herd A and 23-week-old animals of herd B. In most herds, high antibody titres were detected in 4-week-old piglets. These titres decreased during the first 12 weeks of age, but thereafter, increased. It was concluded that PCR with mixed bacterial cultures from tonsillar swabs is a valuable tool for the detection of infected animals. It was also concluded that colonization of tonsils and nasal mucosae can occur in the presence of maternally derived antibodies. Infection of the upper respiratory tract without lung involvement did not result in development of Apx toxin neutralizing antibodies. Therefore, such serological assays cannot be used for the detection of subclinically infected animals.

Polymerase chain reaction optimization for the detection of Pasteurella multocida B:2, the causative agent of Haemorrhagic septicaemia

Specific detection of Pasteurella multocida type B:2 by polymerase chain reaction (PCR), using a set of DNA primers wasoptimised. Effects of the addition of ethylene diamine tetra acetic acid (EDTA) to the sample preparation, Escherichia coli contamination and the number of P. multocida on the PCR product was assessed. The PCR test was compared to the standard bacteriological method for the detection of P. multocida B:2 in tonsillar swab samples collected from slaughter houses of various regions in Indonesia. Addition of 100 mM EDTA-saline to P. multocida B:2 spiked tonsillar swab samples inhibits the production 350 base pairs (bp) PCR product. The inhibitory effect of the EDT A can be eliminated by three times washing with deionised water. The PCR can detect P. multocida as low as I organism and contanimation of 100 CFU of E. coli does not effect the PCR result. The results show that the DNA primers for P. multocida B:2 is sensitive and specific. The inhibitory effect of EDTA in PCR samples can be eliminated by washings. Keywords : PCR, EDT A, Pasteurella multocida B:2

Detection of Actinobacillus pleuropneumoniae in cultures from nasal and tonsillar swabs of pigs by a PCR assay based on the nucleotide sequence of a dsbE-like gene

A PCR assay for the detection of Actinobacillus pleuropneumoniae was developed based on the amplification of a dsbE-like gene. All of 157 field isolates of A. pleuropneumoniae reacted in the PCR by the amplification of a 342 bp product. No reaction was observed with related bacterial species or other bacterial species isolated from pigs, except for A. lignieresii. The lower detection limit of the PCR was 10 2 CFU per PCR test tube and was not affected by the addition of 10 6 CFU Escherichia coli. The PCR was evaluated on mixed bacterial cultures from nasal and tonsillar swabs as well as suspensions of nasal conchae and tonsils obtained from specific pathogen-free (SPF) pigs, experimentally infected pigs, and pigs from farrow-to-finish herds. The results of the new PCR were compared with a PCR based on the detection of the omlA gene coding for an outer membrane protein, with a commercially available PCR (Adiavet™APP, Adiagène, Saint-Brieuc, France), and with conventional culturing. No positive reactions were observed with any of the PCR methods in samples of SPF animals. In samples of the other animals, no or low significant differences between nasal swabs and suspensions as well as tonsillar swabs and suspensions were observed in any method. In general, more positive results were obtained from tonsillar samples in comparison to nasal samples. Interassay sensitivity and specificity values were assessed for each test by pair wise comparisons between assays. The agreement between tests was evaluated by calculating Cohen’s κ coefficient. From these analyses the three PCR assays showed a good agreement. The dsbE-based PCR proved to be highly sensitive (95 and 93%) and specific (82 and 74%) in comparison to the omlA-based PCR and the commercially available PCR, respectively. It was concluded that the dsbE-like gene-based PCR is a reliable diagnostic assay for demonstration of A. pleuropneumoniae. Furthermore, it was demonstrated that tonsillar swabs can be used for the detection of the pathogen in healthy carrier animals.

Naturally occurring tuberculosis in white-tailed deer.

To determine the distribution of lesions and extent of tissues infected with Mycobacterium bovis in a captive population of white-tailed deer. Cross-sectional study. 116 captive white-tailed deer. Deer were euthanatized, and postmortem examinations were performed. Tissues with gross lesions suggestive of tuberculosis were collected for microscopic analysis and bacteriologic culture. Tissues from the head, thorax, and abdomen of deer with no gross lesions were pooled for bacteriologic culture. Tonsillar, nasal, oral, and rectal swab specimens, fecal samples, and samples of hay and pelleted feed, soil around feeding sites, and water from 2 natural ponds were collected for bacteriologic culture. Mycobacterium bovis was isolated from 14 of 116 (12%) deer; however, only 9 of 14 had lesions consistent with tuberculosis. Most commonly affected tissues included the medial retropharyngeal lymph node and lung. Five of 14 tuberculous deer had no gross lesions; however, M bovis was isolated from pooled tissue specimens from the heads of each of these deer. Bacteriologic culture of tonsillar swab specimens from 2 of the infected deer yielded M bovis. Mean (+/- SEM) age of tuberculous deer was 2.5 +/- 0.3 years (range, 0.5 to 6 years). Mycobacterium bovis was not isolated from feed, soil, water, or fecal samples. Examination of hunter-killed white-tailed deer for tuberculosis commonly includes only the lymph nodes of the head. Results of such examinations may underestimate disease prevalence by as much as 57%. Such discrepancy should be considered when estimating disease prevalence.

Human herpesvirus 8 seroprevalence and evaluation of nonsexual transmission routes by detection of DNA in clinical specimens from human immunodeficiency virus-seronegative patients from central and southern Italy, with and without Kaposi's sarcoma.

In order to investigate the seroprevalence of human herpesvirus 8 (HHV-8) infection in central and southern Italy, sera from human immunodeficiency virus (HIV)-seronegative subjects, with and without Kaposi's sarcoma (KS), were analyzed by immunofluorescence assay, using BC-3, a cell line latently infected with HHV-8. High titers of antibody against HHV-8 lytic and latent antigens were detected in all 50 KS patients studied, while in 50 HIV-seronegative subjects without KS, 32 (64%) were found positive for HHV-8 antibodies. Titers in the sera of these patients were lower than those for KS patients. This data suggests that HHV-8 infection is not restricted to KS patients and that the prevalence of HHV-8 infection in the general population may be correlated with differing rates of prevalence of KS in different parts of the world. In view of these findings, possible nonsexual transmission routes were evaluated. Nested PCR was used to test for the presence of HHV-8 DNA in saliva, urine, and tonsillar swabs from KS and non-KS patients. In KS patients, 14 out of 32 tonsillar swabs (43.7%), 11 out of 24 saliva samples (45.8%), and just 2 out of 24 urine samples (8.3%) tested positive for HHV-8 DNA. In the control group, on the contrary, none of the 20 saliva and 20 urine specimens was positive for HHV-8 DNA; only 1 out of 22 tonsillar swabs gave a positive result. This data supports the hypothesis that HHV-8 infects the general population in a latent form. The reactivation of viral infection may result in salivary shedding of HHV-8, contributing to viral spread by nonsexual transmission routes.

Detection of pseudorabies viral DNA in tonsillar epithelial cells of latently infected pigs

SUMMARY The Rice strain of pseudorabies virus (prv) was intranasally instilled in pigs that were seronegative to prv. Cells were scraped or brushed from tonsillar surfaces biweekly until pigs were euthanatized at either 10 or 16 weeks after infection. The dna extracted from tonsillar cells or parenchyma were subjected to polymerase chain reaction analysis, using either a single set of oligonucleotide primers or nested primers from the prv gII glycoprotein gene. Pigs became seropositive to prv by 3 weeks after infection. The virus was isolated from the trigeminal ganglia and tonsils of pigs that were euthanatized or died 1 to 2 weeks after infection, but not from pigs that were euthanatized 10 or 16 weeks after infection. The prv gene products were consistently detected in trigeminal ganglia and tonsils of all pigs at 1, 10, and 16 weeks after infection, and sporadically in the nasal mucosa, lymph nodes, and lungs of pigs that were euthanatized or died during the first 2 weeks after infection. Cells collected biweekly from tonsillar surfaces were mostly nucleated, squamous epithelial cells with fewer lymphocytes and neutrophils. Polymerase chain reaction analysis of dna extracted from these cells revealed prv dna in a large proportion of the samples when sufficient cells were collected to provide 1 μg of extracted dna for use in the reaction mixtures. A second group of pigs had prv strain 4892 intranasally instilled. The virus was isolated from tonsillar swab specimens until 3 weeks after infection. Tonsillar brushing specimens were collected biweekly until 14 weeks after infection. Some brushing specimens contained all nucleated, squamous epithelial cells, whereas other specimens contained a mixture of epithelial cells and up to 15% neutrophils, lymphocytes, and small mononuclear cells. Results of polymerase chain reaction analysis of dna extracted from tonsillar cells collected 5,11, and 14 weeks after infection were consistently positive for prv gene products. Intact cells collected from tonsillar surfaces were placed in polymerase chain reaction mixtures with nested oligonucleotide primers from the prv gII glycoprotein gene and were subjected to multiple amplification cycles. Afterward, the specificity of the amplified prv gene products was determined by hybridization procedures, using a virus-specific oligonucleotide probe. Most nucleated, squamous epithelial cells stained positive for prv dna, suggesting that these cells were the primary source of prv gene products in tonsillar brushing specimens.

Occurrence of Yersinia enterocolitica O:3, Listeria monocytogenes O:4 and thermophilic Campylobacter spp. in slaughter pigs and carcasses in Trinidad

Pigs slaughtered at an abattoir in Trinidad were sampled for the occurrence of Yersinia, Listeria and Campylobacter at different sites. The antibiograms of Yersinia isolates were also determined. Of 141 samples each of rectal and tongue swabs taken, 23(16·1%) and 9(6·4%) were positive for Y. enterocolitica compared to 11(7·3%) of 150 tonsillar samples yielding the organism. Rectal swabs had a statistically significantly ( P ≤ 0·05, x 2 ) higher frequency of isolation than tongue or tonsillar swabs. All 43 isolates of Y. enterocolitica were serotype O:3 biotype 4. From a total of 139 rectal swabs cultured for thermophilic Campylobacter , 75(51·8%) were C. coli and 3(2·2%) were positive for C. jejuni . The difference was statistically significant ( P ≤ 0·001; x 2 ). Carcass swabs also yielded significantly more C. coli , 27(17·4%) than C. jejuni , 4(2·6%) of 155 samples cultured. Seven (5·0%) of 139 rectal swabs tested were positive for Listeria monocytogenes while only 3(1·9%) of 155 carcass samples yielded the organism but the difference was not statistically significant ( P ≥ 0·05; x 2 ). Nine of 10 isolates of L. monocytogenes were serotype 4 while one was serotype 1. Resistance of the 43 isolates of Y. enterocolitica was highest to penicillin (100·0%), neomycin (100·0%) but least to gentamycin (2·3%) and sulfamethoxazole/trimethoprim (0·0%). It was concluded that Y. enterocolitica serotype O:3, biotype 4 is predominant in pigs in Trinidad and the isolation of L. monocytogenes , particularly serotype 4, and C. coli and C. jejuni from pork carcasses may pose a health risk to consumers.

The role of Chlamydia pneumoniae in severe acute tonsillitis.

Chlamydia pneumoniae has been implicated as a cause of tonsillitis and pharyngitis, but the incidence has varied from one to 19 per cent in various studies. We investigated 51 patients admitted to University College Hospital, Galway, with severe tonsillitis. Throat swabs were examined for evidence of Chlamydia pneumoniae using a direct monoclonal antibody test. Blood was taken for serology from 45 patients. A further specimen was taken at six weeks. A control group of 32 blood bank sera was used. Mean hospital stay was three days (one to eight). Five patients (10 per cent) were monospot positive. Chlamydia pneumoniae was identified by direct immunofluorescence on a tonsillar swab from one patient who did not seroconvert. IgG antibody was identified in 13 cases (29 per cent) and in seven of the control group (22 per cent). No serological evidence of recent infection was found. Chlamydia pneumoniae was not found to be a cause of severe acute tonsillitis in our study group.

Bacteremia During Tonsillectomy

FRANCOIS, M.; BINGEN, E. H.; LAMBERT-ZECHOVSKY, N. Y.; MARIANI-KURKDJIAN, P.; NOTTET, J -B.; NARCY, P. Author Information

Bacteremia during tonsillectomy.

To determine the risk of bacteremia during tonsillectomy, we cultured blood specimens that were taken from 32 children during surgery and tonsillar swabs that were obtained just before excision, and compared the results with quantitative cultures of the excised tonsillar tissue. Twenty-five children had Haemophilus influenzae within the tonsillar tissue (density range, 10(3) to 10(8) colony-forming units per gram), and seven had Streptococcus pyogenes (density, 10(3) colony-forming units per gram in one case, 10(5) colony-forming units per gram in one case, and 10(6) colony-forming units per gram in five cases). Twelve perioperative blood cultures were positive; H influenzae was found nine times, and Micrococcus species was found one time, and alpha-hemolytic streptococci were found two times. Haemophilus influenzae was always present in the corresponding tonsillar specimens, although there was no apparent relationship between the density of colonization of the tonsillar tissue and a positive blood culture.

Isolation of Chlamydia pneumoniae and antibodies to the agent in patients with acute bronchitis

Isolation of Chlamydia pneumoniae and serologic tests were performed on 85 children and adults patients with acute bronchitis admitted to the Kitasato Institute Hospital from January 1989 to May 1991. C. pneumoniae was isolated from sputa or tonsillar swabs in 11 of 57 (19%) patients examined. Of these, 10 patients were tested serologically and 9 (90%) had 16 or higher titer of IgG antibody. A total of 68 patients were tested serologically, and 17 (25%) revealed positive; 14 had a fourfold rise or more in the titer of IgG antibody, or IgM titer of 16 or more, and 3 had IgG antibody with a titer of 512 or greater. In 34 patients, sera were tested for evidence of acute infections not only with C. pneumoniae, but also with Mycoplasma pneumoniae, influenza A virus, influenza B virus, adenovirus, respiratory syncytial virus, coxackie A9 virus, parainfluenza virus type I, II, and III. Two of the 34 patients were associated with influenza A virus, 4 with influenza B virus, whereas 6 were associated with C. pneumoniae. The data available suggest that C. pneumoniae cause bronchitis much more than has been suspected, and also confirm earlier suggestions that the agent played an important role in respiratory tract infections. Chlamydia trachomatis was recovered from sputa and tonsillar swabs in 6 adults patients with acute bronchitis. Studies are needed for a better understanding of the epidemiology and clinical relevance of this microorganism to lower respiratory tract diseases in adults.

Experimental establishment of persistent infection in swine with a zoonotic strain of Salmonella newport

SUMMARY An experiment was conducted to determine whether a persistent Salmonella newport infection could be established in swine, to determine duration of shedding and distribution of the organism in internal organs, and to determine whether changes occurred in antimicrobial susceptibility or plasmid profile of the organism during the course of long-term infection. Naturally farrowed Salmonella-free pigs (n = 22) were orally exposed to a multiply antimicrobial-resistant zoonotic strain of S newport when they were 7 weeks old. Tonsillar and rectal swab specimens were examined bacteriologically for S newport during the first week after exposure, then weekly for 7 weeks. Fecal samples were likewise examined weekly or every 2 weeks for 28 weeks after exposure. Necropsies of 2 or 3 randomly selected pigs were conducted at 2, 4, 8, 12, 16, 20, 24, and 28 weeks after exposure. A total of 45 specimens/pig representing the following internal organs or tissues were examined bacteriologically for S newport: liver, spleen, kidney, gallbladder, heart, heart blood, lung, stomach, and tonsils; segments of the intestinal tract with corresponding lymph nodes; and lymph nodes from lymphocenters of the head and neck, thoracic cavity, thoracic limbs, abdominal viscera, and abdominal wall. Exposure to S newport induced a mild and transient clinical response. The organism was recovered from 97% of tonsillar swab specimens and 89% of rectal swab specimens collected during 7 weeks after exposure and from 98% of fecal samples collected during 28 weeks after exposure. At necropsy, S newport was recovered most frequently from tonsils (86.4%), followed by segments of the intestinal tract from ileum to rectum (81.8% recovery from cecal contents), and from mandibular (68.2%), jejunal (50%), and ileocolic (45.5%) lymph nodes. Sporadic recoveries of the organism were made from other lymph nodes and from gallbladder, stomach, kidney, spleen, liver, and heart, varying from 2 to 20 weeks after exposure. The cranial portion of jejunum, medial iliac lymph nodes, dorsal superficial cervical lymph node, and heart blood of all pigs were culture-negative. Of 26 representative isolates of S newport recovered from body organs or feces during 28 weeks after exposure, 4 (15.4%) underwent changes in antimicrobial susceptibility pattern. Changes in plasmid profile of the organism were not detected during longterm infection of swine.

DETECTING NONHEMOLYTIC PASTEURELLA HAEMOLYTICA INFECTIONS IN HEALTHY ROCKY MOUNTAIN BIGHORN SHEEP (OVIS CANADENSIS CANADENSIS): INFLUENCES OF SAMPLE SITE AND HANDLING

Effects of sampling procedures on ability to culture Pasteurella spp. from Rocky Mountain bighorn sheep (Ovis canadensis canadensis) were examined experimentally. Sample site influenced (P less than 0.0001) recovery of P. haemolytica in adult bighorn sheep. We isolated nonhemolytic P. haemolytica from 18 of 19 tonsillar swabs and 18 of 19 tonsillar biopsies from adult sheep, yet only four of 19 nasal swabs yielded isolates. Sample handling also affected (P less than 0.0001) recovery of P. haemolytica. Nonhemolytic P. haemolytica was cultured from 14 of 19 tonsillar swabs plated directly onto blood agar, but from only two of 19 swabs stored for 24 hr in modified Stuart's medium. We detected nonhemolytic P. haemolytica at least once in bronchial aspirates from four and in nasal swabs from three of six bighorn lambs. Based on direct cultures of tonsillar swabs and/or biopsies, all 26 bighorn sheep (seven lambs, 19 adults) sampled were infected with nonhemolytic P. haemolytica; only two lambs developed pneumonia during the study period. Thirty-four of 37 nonhemolytic P. haemolytica isolates tested were biotype T; three were biotype A. Serotypes 3; 4; 3, 4 and 3, 4, 10 were identified in a subsample of 17 isolates. Our data suggest tonsillar swabs or biopsies plated directly onto blood agar and incubated immediately offer the greatest probability of recovering nonhemolytic P. haemolytica from health bighorn sheep.

&lt;I&gt;Chlamydia pneumoniae&lt;/I&gt;と&lt;I&gt;Chlamydia trachomatis&lt;/I&gt;が検出された滲出性中耳炎, 気管支炎を併発した習慣性扁桃炎症例

We report a case of recurrent tonsillitis and otitis media with effusion (OME) from which Chlamydia trachomatis was isolated. Chlamydia pneumoniae, a newly recognized species of Chlamydia, was also recovered from the tonsillar and bronchial swabs. A 8-year-old girl was seen on February 23, 1988, because of a running nose, a productive cough and bilateral hearing difficulty. She had a history of recurrent tonsillitis. The diagnosis was acute sinusitis with tubal obstruction, then cefixime was prescribed. Her symptoms were once resolved, for the time being but she came back to the hospital a week later with a bilateral ear-ache. The tympanic membranes were injected and characteristically retracted. Her left ear showed type B tympanogram (effusion). Tympanocentesis was performed to remove middle-ear effusion, from which C. trachomatis but no ordinary bacterium was isolated. Therefore rokitamycin 300 mg/day was administered for a week. Her condition improved, however, a rhinorrhea, a plugged ear sensation and a hacking cough returned in a month. She was admitted to the hospital on May 10, for tympanostomy and grommet insertion, but from the day before admission, she had a sore throat with fever (39.2 degrees C). The surgery was withheld until May 26. When adenotonsillectomy and grommets insertion were undertaken, C. trachomatis had disappeared from the middle-ear effusion, but C. pneumoniae was recovered from both tonsillar and bronchial swabs. Readministration of rokitamycin was performed and to date (June, 1990) she remains well.

Ｃｈｌａｍｙｄｉａ ｐｎｅｕｍｏｎｉａｅによる気道感染症

A newly recognized chlamydial species, Chlamydia pneumoniae causes acute respiratory infections including pneumonia, bronchitis and pharyngitis. In this paper, eight cases of bronchitis and tonsillitis associated with C. pneumoniae are presented. Three cases came to the clinic because of persistent cough and productive sputum. C. pneumoniae was isolated from sputum of a patient and cultured in HeLa 229 cells. Other two patients were diagnosed serologically; Antibodies were measured by microimmunofluorescence using formalized elementary bodies of C. pneumoniae. A titer of 512 in the IgG class was detected. Four patients had sore throat. C. pneumoniae was isolated and cultured from tonsillar swabs in all of them. A patient with sore throat and cough diagnosed as pharyngolaryngitis was sero-positive. Antibodies to C. pneumoniae in IgG and IgM class were 128 and 32, respectively. All the patients were treated with macrolide antibiotics (erythromycin and rokitamycin), and clinical symptoms subsided. In five patients from whom the organism was isolated, the agents were eradicated by the treatment. However, clinical courses of those patients revealed that patient takes a long time to recover from the illness, if diagnosis and first choice of antimicrobial agent are not appropriate.

Changes in tonsillar bacteriology of recurrent acute tonsillitis: 1980 vs. 1989

Recurrent acute tonsillitis is a common problem. Despite this, there still remain many controversies regarding aetiology and correct management. The tonsillar microflora of 33 children with recurrent acute tonsillitis studied in 1980 and 58 patients studied in 1989 is presented. A comparison of the microbiology in the two periods studied a decade apart suggests that the pathogenic profile is changing. Haemophilus influenzae increased from 39 to 62% in the deep tonsillar tissue in the decade. There was a concomitant increase in incidence of Staphylococcus aureus from 6 to 40% of cases. In the same interval, mixed microflora increased from 18 to 52%. Anaerobic organisms were isolated in insignificant numbers. Unique to this study, 44% of H. influenzae isolates in 1989 were beta lactamase producers, increasing from only 2% in 1980. All of the S. aureus were beta lactamase producers. In the majority, the throat swabs grew only organisms commensal to the upper respiratory tract however, the deep tonsillar tissue excised at tonsillectomy carried significant growths of pathogenic organisms confirming the inadequacy of the superficial tonsillar swab as an indicator of treatment.