Detection of strangles carriers.
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Animals↗
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Molecular basis of variation in protective SzP proteins of Streptococcus zooepidemicus.
OBJECTIVES: To characterize, on a molecular basis, variable regions of the SzP proteins of the Moore and Bryans serovars of Streptococcus zooepidemicus and specificity of opsonic responses. SAMPLE POPULATION: 14 Moore and Bryans serovars of S zooepidemicus. PROCEDURE: Using polymerase chain reaction analysis and primers from the 5' and 3' sequences of the prototype gene SzPW60, the SzP genes of each Moore and Bryans serovar were sequenced and translated, then the amino acid sequences were compared. RESULTS: Comparison of the amino acid sequences revealed 2 variations at the N terminus; a hypervariable (HV) region from residue 106 to 166, approximately; and proline-glutamic acid-proline-lysine repeats in the carboxy terminus that ranged in number from 7 to 12. Five distinct motifs, HV 1 to 5, which varied independently of the N termini were found in the internal HV region. All serovars were opsonized by antiserum to the prototype SzPW60 protein, indicating that opsonogenic epitopes are on the conserved regions of the protein. CONCLUSION AND CLINICAL RELEVANCE: Variant motifs may be valuable in epizootiologic and pathogenesis studies of S zooepidemicus infections of the respiratory tract of young horses and in determining whether there are populations of S zooepidemicus unique to specific animal hosts. It is also clear from the opsonic responses to SzP that at least a portion of the protective responses are probably not serovar specific.
Amino Acid Sequence↗
Use of the protective antigen of Erysipelothrix rhusiopathiae in the enzyme-linked immunosorbent assay and latex agglutination.
To establish a safe and convenient serodiagnostic method for swine erysipelas, a purified protective protein antigen of Erysipelothrix rhusiopathiae, which included a large amount of protective protein (64 kDa protein), was used for enzyme-linked immunosorbent assay (ELISA) and the latex agglutination (LA) test. In the ELISA, the antisera to four different serovars (1a, 2, 5 and 20) of E. rhusiopathiae exhibit a positive reaction, while antisera to other species of bacteria (Listeria monocytogenes, Staphylococcus aureus, Streptococcus suis, Rhodococcus equi and Corynebacterium pseudotuberculosis) exhibit a negative reaction. In the LA test, the antisera to three different serovars (1a, 2 and 5) of E. rhusiopathiae reacted with P64-sensitized latex beads, while the antiserum to serovar 20 (2553 strain) did not. Moreover, the antisera to other species of bacteria (Listeria monocytogenes, Staphylococcus aureus, Streptococcus suis, Rhodococcus equi and Corynebacterium pseudotuberculosis) did not in this test. Comparing the results of the growth agglutination (GA), ELISA and LA tests of 284 swine sera, there was a high degree of correlation among the results. The detection of anti-E. rhusiopathiae antibodies in the GA, ELISA and LA tests were compared using sera from pigs immunized with P64, alkaline extract (AE) and live-cell vaccine (LV). In all three tests, anti-E. rhusiopathiae antibodies could be detected 1 week after immunization. The serum antibody titre as determined by the LA test increased moderately, as did that by the GA test, while that determined by ELISA increased rapidly. These results suggested that ELISA could be used to monitor changes in anti-E. rhusiopathiae antibody titre and the LA test could be used in the screening test for swine erysipelas.
Animals↗
Pharmacokinetics and therapeutic potential for repeated oral doses of trimethoprim/sulphachlorpyridazine in horses.
The pharmacokinetic parameters of a powder formulation of trimethoprim/sulphachlorpyridazine were studied in eight healthy horses which received 5 mg/kg trimethoprim and 25 mg/kg sulphachlorpyridazine 12-hourly with concentrate for five days. The intake of the medicated concentrate by the horses was variable during the first two days, but after they became accustomed to the taste the intake by all the horses during the last three days was good. Faecal samples taken before and on the last day of the drug administrations were negative when cultured for salmonella. Compared with the results of a previous single-dose experiment, higher plasma concentrations and a higher area under the curve for both the drugs were observed. The repeated doses provided plasma concentrations above the minimal inhibitory concentration for Streptococcus zooepidemicus, S equi, Actinobacillus equuli and Rhodococcus equi isolated from the respiratory tract of horses. Synergism between the two drugs occurred at different drug concentration ratios with different bacterial species.
Administration, Oral↗
[The first isolation of Dermatophilus congolensis van Saceghem 1913 in Czechoslovakia].
The authors present a report on the first detection of the strain Dermatophilus congolensis in Czechoslovakia. The strain was isolated from ulcerations on the dorsal part of the foot of a 47-year-old patient who previously worked for several years in Libya. Based on examinations of smears from cultures of an isolated strain and four strains from a collection, from experimental dermatofilosis of a rabbit the authors describe the micromorphology of strains of D. congolensis under a light and electron microscope (transmission an and raster microscope). They describe the colonial morphology, haemolytic properties and in particular detail the haemolytic interaction of strains of D. congolensis with Streptococcus agalactiae, Rhodococcus equi and with strains of S. aureus (monoproducers of beta-haemolysin and delta-haemolysin resp.). They assessed the sensitivity of strains of D. congolensis to some staphylococcal bacteriocin-like substances.
Actinomycetales↗
Characterization and utilization of monoclonal antibodies reactive to Yersinia pseudotuberculosis.
The 3 murine monoclonal antibodies, Yps1, Yps2 and Yps3 reactive to Y. pseudotuberculosis can be stabilized and all were found to be of IgG type. Monoclonal antibody, Yps1, recognized a glycoprotein antigen of the organism with reactivity at the 55-75 kDa region, while Yps2 and Yps3 recognized protein antigens of Y. pseudotuberculosis 65 kDa and 26-28 kDa molecular weight regions, respectively. The specificity of monoclonal antibodies was tested using dot ELISA and Western blotting with whole cell organisms or whole cell sonicated soluble antigens of different Yersinia species, Salmonella typhi, Klebsiella pnemoniae, Streptococcus abortus-equi and Escherichia coli. Monoclonal antibody, Yps1 exhibited cross-reactivity with soluble antigens and whole cell preparations of Y. pestis. Yps2 cross-reacted to soluble antigens of all the tested bacteria. Reactivity of monoclonal antibody, Yps3 was restricted to Y. pseudotuberculosis and Y. pestis with soluble antigen preparations. No reaction was observed with Yps2 and Yps3 to whole cell organism preparations from tested bacteria including Y. pseudotuberculosis. The co-agglutination reagent prepared by sensitizing staphylococcal cells with Yps1 monoclonal antibody produced a positive agglutination with all the 4 Y. pseudotuberculosis isolates and the 3 Y. pestis strains tested. Sandwich dot ELISA using monospecific antisera as a capture antibody and a monoclonal antibody, and Yps3 as a revealing antibody had a high level of specificity in detecting Y. pseudotuberculosis antigens.
Animals↗
Phospholipase C in Listeria.
The cooperative and antagonistic effect of extracellular bacterial proteins of Streptococcus agalactiae, Rhodococcus equi, Corynebacterium ovis, and Corynebacterium haemolyticum with proteins of listerial strains of various sources, species and serovars of the membrane of sheep erythrocytes was investigated. Listeria monocytogenes and Listeria ivanovii produce, beside listeriolysin, further proteins. Their specific effect on the sheep erythrocyte membrane becomes apparent after the appearance of substances produced by the organisms under study. Ten strains of L. ivanovii produced phospholipase C responsible for the zone of incomplete haemolysis. It was inhibited by the non-haemolytic sphingomyelinase D of C. ovis and C. haemolyticum. Chromatographic analysis revealed that phospholipase C splits sphingomyelin in the membrane of sheep erythrocytes. The inhibition of double haemolysis of L. ivanovii by sphyngomyelinase D of C. ovis on agar plates with washed (!) sheep erythrocytes can be utilized as a specific and rapid identification test of L. ivanovii.
Animals↗
Identification & characterisation of the two novel streptococcal pyrogenic exotoxins SPE-L & SPE-M.
BACKGROUND & OBJECTIVES: The streptococcal pyrogenic exotoxins (SPEs) are produced by Streptococcus pyogenes and belong to the family of bacterial superantigens, a group of highly mitogenic proteins. The aim of this study was to search unfinished streptococcal genomes for novel superantigens, to generate recombinant proteins from potential open reading frames (ORFs) and to analyse them for superantigen activity. METHODS: The microbial genome database was searched using a TBLASTN search programme. Genotyping of S. equi and S. pyogenes isolates was done using the specific primer pairs. The spe-l and spe-m genes were amplified by PCR. RESULTS: Two novel streptococcal superantigen genes (sepe-l and sepe-m) were identified from the Streptococcus equi genomic database at the Sanger Centre. Genotyping of S. pyogenes isolates resulted in the detection of the orthologous genes spe-l and spe-m in a restricted number of S. pyogenes isolates and revealed a link of spe-l to the M89 serotype. Recombinant SPE-L and rSPE-M were highly mitogenic for human peripheral blood lymphocytes with half maximum responses at 1 pg/ml and 10 pg/ml, respectively. The results from competitive binding experiments suggest that both proteins bind MHC class II at the beta-chain, but not at the alpha-chain. The most common target for both toxins were human Vbetal.1 expressing T cells. Seroconversion against SPE-L and SPE-M was observed in healthy blood donors. INTERPRETATION & CONCLUSION: The two novel ORFs identified in both, S. equi and S. pyogenes, code for proteins that show typical superantigen features. The seroconversion seen in some blood donors suggest that the proteins are indeed produced by the bacteria. Interestingly, the spe-l gene is highly associated with S. pyogenes M89, which is linked to acute rheumatic fever in New Zealand.
Bacterial Proteins↗
Severe Streptococcus infection in spotted hyenas in the Ngorongoro Crater, Tanzania.
In a population of spotted hyenas (Crocuta crocuta) monitored between 1996 and 2005 in the Ngorongoro Crater, Tanzania, 16 individuals from five of eight social groups displayed clinical signs of an infection, including severe unilateral swelling of the head followed by abscess formation at the mandibular angle, respiratory distress, mild ataxia, and lethargy. Two (12.5%) of these 16 individuals died within days of developing signs. Clinical signs in hyenas were first noted in 2001, and most cases occurred between September 2002 and February 2003, suggesting an outbreak of infection during this period. Histopathological examination of internal organs from one hyena that died with signs revealed morphological changes consistent with severe bacterial infection. Phenotypic examination and phylogenetic analysis of the 16S rRNA gene of the causative agent of infection revealed a Lancefield group C Streptococcus with a high level of homology to S. equi subsp. ruminatorum, a subspecies of S. equi recently described in domestic sheep (Ovis aries) and goats (Capra hircus) with mastitis in Spain. Strains similar to this bacterium were also isolated from two hyenas without obvious clinical signs, suggesting that hyenas may be 'carriers' of this bacterium, and from a sympatric Burchell's zebra (Equus burchelli), a herbivore species often consumed by hyenas. To our knowledge this is the first report of a Streptococcus infection in these two wildlife species. The high genetic similarity between the hyena and zebra isolates indicates that inter-specific transmission may occur, possibly when hyenas consume infected zebra carcasses.
Animals↗
Group-C beta-hemolytic streptococcal bacteremia.
Group-C beta-hemolytic streptococci (GCBHS) is an uncommon cause of bacteremia. In a 5-year period, GCBHS accounted for 0.28% of positive blood cultures and 0.35% of bacteremias documented at our hospital. The incidence of GCBHS bacteremia was 0.05 episodes per 1000 admissions. We were able to analyze clinical data of 10 of the 13 patients with GCBHS bacteremias. All but one were adults with significant underlying diseases, and seven episodes were community acquired. The skin was the portal of entry in only one case. Clinical syndromes included primary bacteremia (four cases), pneumonia (two cases), endocarditis (two cases), and meningitis, intraabdominal infection, and metastatic suppurative pericarditis (one episode each). Of 13 isolates, 12 were identified to species level: six, Streptococcus equisimilis; three S. equi; two S. dysgalactiae; and one S. zooepidemicus. Resistance to penicillin was detected in one isolate and none of our isolates displayed penicillin tolerance, Four patients died (40%) despite appropriate antimicrobial therapy.
Adolescent↗
Routine use of PCR-restriction fragment length polymorphism analysis for identification of mycobacteria growing in liquid media.
A PCR-restriction fragment length polymorphism (PCR-RFLP) procedure capable of rapidly identifying 28 species of clinically encountered mycobacteria was evaluated for use in the routine identification of acid-fast isolates growing in BACTEC 12B and 13A liquid media. PCR-RFLP identified 100 of 103 acid-fast isolates recovered from 610 patient specimens submitted for culture during the study. The three isolates unidentifiable by PCR-RFLP produced restriction patterns not included in the PCR-RFLP algorithm and could therefore not be assigned to a species. These isolates were characterized by their morphologic and biochemical characteristics. Two of the isolates were identified as M. terrae complex and M. gordonae. The third isolate could not be definitively identified and could only be characterized as a Mycobacterium sp. most closely resembling M. chelonae. PCR-RFLP identifications agreed with the conventional identifications for 96 of the 100 isolates identified by PCR-RFLP. Subsequent identification of the four discordant isolates by gas chromatography analysis supported the PCR-RFLP identification of each isolate. Amplification products were also obtained from isolates of Streptococcus albus and Rhodococcus equi recovered from patient specimens; however, the restriction patterns of these nonmycobacterial species did not resemble the patterns of any mycobacterial species included in the PCR-RFLP algorithm. PCR-RFLP seems to be a reliable procedure for the routine identification of mycobacteria and has the potential for providing identifications of mycobacterial isolates which are more accurate than conventional identification techniques based on morphologic and biochemical characteristics.
Bacterial Typing Techniques↗
The pathogenic equine streptococci.
Streptococci pathogenic for the horse include S. equi (S. equi subsp. equi), S. zooepidemicus (S. equi subsp. zooepidemicus), S. dysgalactiae subsp. equisimilis and S. pneumoniae capsule Type III. S. equi is a clonal descendent or biovar of an ancestral S. zooepidemicus strain with which it shares greater than 98% DNA homology and therefore expresses many of the same proteins and virulence factors. Rapid progress has been made in identification of virulence factors and proteins uniquely expressed by S. equi. Most of these are expressed either on the bacterial surface or are secreted. Notable examples include the antiphagocytic SeM and the secreted pyrogenic superantigens SePE-I and H. The genomic DNA sequence of S. equi will greatly accelerate identification and characterization of additional virulence factors and vaccine targets. Although it is the most frequently isolated opportunist pyogen of the horse, S. zooepidemicus has been the subject of few contemporary research studies. Variation in the protectively immunogenic SzP proteins has, however, been well characterized. Given its opportunist behavior, studies are urgently needed on regulation of virulence factors such as capsule and proteases. Likewise, information is also very limited on virulence factors and associated gene regulation of S. dysgalactiae subspecies equisimilis. It has recently been shown that equine isolates of Streptococcus pneumoniae are clonal, a feature shared with S. equi. All equine isolates express capsule Type III, are genetically similar, and have deletions in the genes for autolysin and pneumolysin. In summary, the evolving picture of the interaction of the equine pathogenic streptococci and their host is that of multiple virulence factors active at different stages of pathogenesis. The inherent complexity of this interaction suggests that discovery of effective combinations of immunogens from potential targets identified in genomic sequence will be laborious.
Animals↗
[Other infections (Streptococcus pneumoniae, Haemophilus influenzae, Pseudomonas aeruginosa, Salmonella spp., Campylobacter spp., Nocardia asteroides, Rhodococcus equi and Bartonella spp.)].
People infected with human immunodeficiency virus (HIV) are at increased risk for bacterial infections due to HIV-associated immunologic defects. Bacterial infections were found to be, both a predictor of progression to AIDS and a substantial cause of mortality in pre-AIDS stages. Most bacterial infections are caused by Streptococcus pneumoniae, Haemophilus influenzae, Salmonella spp. and Pseudomonas aeruginosa. Rhodococcus equi, Nocardia spp., Campylobacter spp. and Bartonella spp. are less common. Data derived from two AIDS Clinical Trials Group studies showed that the most common bacterial infections were sinusitis (8.5 per 100 episodes per person years [py]), bacterial pneumonia (5.0 per 100 py), bronchitis (4.1 per 100 py) and soft tissue infections (3.5 per 100 py). In this review clinical characteristics and treatment recommendations according to data available in the literature for these infections are summarized.
AIDS-Related Opportunistic Infections↗
Report of cases of and taxonomic considerations for large-colony-forming Lancefield group C streptococcal bacteremia.
Traditionally, group C streptococci include four species: Streptococcus equisimilis, S. zooepidemicus, S. equi, and S. dysgalactiae, the first three of which are group C beta-hemolytic streptococci (GCBHS). However, many of the beta-hemolytic streptococci carrying Lancefield group C antigen isolated from clinical specimens are S. milleri. These organisms can be differentiated by colony size. We retrospectively collected data concerning large-colony-forming GCBHS bacteremia that occurred during a period of 8 years at the Massachusetts General Hospital. A total of 222 cases of beta-hemolytic streptococcal bacteremia were identified; data on the Lancefield grouping were available in 192 cases: 45 cases (23.6%) were group A, 96 cases (50%) were group B, 7 cases (3.6%) were group C (large colony forming), and 44 cases (22.9%) were group G. The medical records for cases of large-colony-forming GCBHS bacteremia were reviewed. In one case, the isolate was thought to be a contaminant; the other six cases are reported (five males and one female; mean age, 55 years). All patients had severe underlying conditions, and none had a history of exposure to animals. The clinical syndromes included two cases of cellulitis and one case each of endocarditis, myocardial infarction complicated by infection, pneumonia, and myofasciitis. The diagnoses for two patients with endovascular infections were delayed. Three of the six patients had fatal outcomes, and other two, after prolonged hospitalization, were transferred to a long-term rehabilitation center. We concluded that the severe outcomes reflect delay in diagnosis and treatment as well as the severity of the underlying diseases. The taxonomy of GCBHS is discussed. More reports differentiating large- and small-colony-forming GCBHS are needed.
Adult↗
Frozen cancellous bone allografts: positive cultures of implanted grafts in posterior fusions of the spine.
We have carried out a study on the behaviour pattern of implanted allografts initially stored in perfect conditions (aseptically processed, culture-negative and stored at -80 degrees C) but which presented positive cultures at the implantation stage. There is no information available on how to deal with this type of situation, so our aim was to set guidelines on the course of action which would be required in such a case. This was a retrospective study of 112 patients who underwent a spinal arthrodesis and in whom a total of 189 allograft pieces were used. All previous bone and blood cultures and tests for hepatitis B and C, syphilis and HIV (via PCR techniques) were negative. The allografts were stored by freezing them at -80 degrees C. A sample of the allograft was taken for culture in the operating theatre just before its implantation in all cases. The results of the cultures were obtained 3-5 days after the operation. There were 22 allografts with positive culture results (12%) after implantation. These allografts were implanted in 16 patients (14%). Cultures were positive for staphylococci coagulase negative (ECN) in 10 grafts (46%), Pseudomonas stutzeri in two grafts (9%), Corynebacterium jeikeium in two grafts (9%), staphylococci coagulase positive in two grafts (9%) and for each of the following organisms in one case each (4%): Corynebacterium spp., Actinomyces odontolyticus, Streptococcus mitis, Peptostreptococcus spp., Rhodococcus equi and Bacillus spp. No clinical infection was seen in any of these patients. Positive cultures could be caused by non-detected contamination at harvesting, storing or during manipulation before implantation. The lack of clinical signs of infection during the follow-up of our patients may indicate that no specific treatment different from our antibiotic protocol is required in the case of positive culture results of a graft piece after implantation.
Adolescent↗
Bronchointerstitial pneumonia and respiratory distress in young horses: clinical, clinicopathologic, radiographic, and pathological findings in 23 cases (1984-1989).
Twenty-three foals, between 1 and 7 months old, with signs of acute respiratory distress, were examined at the Veterinary Medical Teaching Hospital (VMTH), University of California, Davis, between 1984 and 1989. Characteristic features included sudden onset of severe respiratory distress and tachypnea, cyanosis unresponsive to nasal oxygen, pyrexia, hypoxemia, hypercapneic respiratory acidosis, poor response to treatment, and histopathologic lesions of bronchiolitis and bronchointerstitial pneumonia. Seven of the 23 foals were normal before the onset of respiratory distress, 3 foals were found dead, and 13 foals were being treated for respiratory tract infections at the time of presentation. Laboratory data obtained for 13 horses showed increased plasma fibrinogen concentration (630.7 +/- 193 mg/dL), leukocytosis (18,607 +/- 7,784/microL), and neutrophilia (13,737 +/- 8,211/microL). Thoracic radiographs showed a diffuse increase in interstitial and bronchointerstitial pulmonary opacity and, in 5 foals, an alveolar pulmonary pattern of increased density was also seen. In 3 foals heavy interstitial infiltration proceeded to a coalescing nodular radiographic appearance. Microbiological culture of tracheobronchial aspirates (TBA) from 9 foals yielded bacterial growth, but no one bacterial species was consistently isolated. Microbiological culture of postmortem specimens of the lung from 6 foals yielded growth of bacteria that included Escherichia coli, Enterobacter spp., Proteus mirabilis, Klebsiella pneumoniae, Rhodococcus equi, or beta-hemolytic Streptococcus spp. Tracheobronchial aspirates from 4 foals and lung samples collected from a further 4 foals at necropsy yielded no bacterial growth. Cultures were not taken from two foals premortem or postmortem. Virologic examination of TBA, lung tissue, or pooled organ tissue from 12 foals was negative. Viral culture of TBA from 1 foal showed cytopathic effects and positive immunofluorescence for equine herpes virus type II (EHV-II). In addition to the 3 foals that were found dead, 11 foals died or were euthanatized. Pathologic lesions were limited to the lungs in 50% of the foals; the remainder also had bowel lesions suggestive of hypoxic injury. The predominant histopathologic pulmonary lesions included bronchiolitis, bronchiolar and alveolar epithelial hyperplasia, and necrosis. Many bronchioles were filled with mucoid and fibrinocellular exudate. The peribronchiolar interstitium and adjacent alveolar spaces were also infiltrated with inflammatory cells and contained proteinaceous edema fluid. Type II cell hyperplasia and hyaline membrane formation were observed in the majority of foals and in 2 foals alveolar multinucleate giant cells were also present.(ABSTRACT TRUNCATED AT 400 WORDS)
Animals↗
Review of 40 cases of lung abscesses in foals and adult horses.
A retrospective study of 40 horses with primary lung abscesses without pleuropneumonia showed that lung abscesses occurred predominantly in young horses, usually in foals aged < or = 6 months. Hyperfibrinogenaemia was present in all but 2 horses, and other common clinical and haematological findings were hyperthermia, tachycardia, polypnoea, and neutrophilic leucocytosis. The most common bacterial species isolated from transtracheal aspirates were Streptococcus zooepidemicus (20/34) and Rhodococcus equi (13/34). Most horses (23/27) discharged from the hospital were long term survivors. Of the 19 horses for which a follow up was available, 12 raced or performed the task for which they were intended. None of the indicators evaluated in this study could reliably predict the bacterial species involved or the outcome.
Actinomycetales Infections↗
Pulsed-field gel electrophoresis is more discriminating than multilocus enzyme electrophoresis and random amplified polymorphic DNA analysis for typing pyogenic streptococci.
The SmaI restriction endonuclease digestion patterns of chromosomal DNAs from 99 pyogenic streptococci belonging to Lancefield group A (41 Streptococcus pyogenes), group C (seven S.dysgalactiae, 11 "S. equisimilis", three S. equi, eight S. zooepidemicus) and group G (25 human group G Streptococcus, four S. canis) were analyzed by pulsed-field gel electrophoresis (PFGE), and the results were compared with those previously obtained by multilocus enzyme electrophoresis (MLEE) and random amplified polymorphic DNA analysis (RAPD). PFGE revealed 93 distinct types among the 99 strains, and no patterns were common to strains of different species. The discriminatory power of PFGE was greater than that of MLEE and RAPD for groups A and G streptococci. The polymorphism among group C streptococci was similar with the three techniques. PFGE is, therefore, the most efficacious method for epidemiological typing of pyogenic streptococci.
DNA, Bacterial↗