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PubMed2003

Production and properties of bacteriocin-like inhibitory substances from the swine pathogen Streptococcus suis serotype 2.

Streptococcus suis serotype 2 is a major pathogen found in the upper respiratory tract of swine. In this study, isolates of this bacterial species were tested for the production of bacteriocin-like inhibitory substances (BLIS). Of the 38 strains tested, four inhibited the growth of other S. suis isolates according to a deferred-antagonism plate assay. Interestingly, three of the strains were originally isolated from healthy carrier pigs and were considered nonvirulent. Three isolates (94-623, 90-1330, and AAH4) that produced BLIS in liquid broth were selected for further characterization. None of the inhibitory activities was related to the production of either organic acids or hydrogen peroxide. The BLIS produced by these strains were heat stable and proteinase K, pronase, and elastase sensitive but were trypsin and chymotrypsin resistant. They were stable at pH 2 and 12 and had molecular masses in the range of 14 to 30 kDa. Maximum production was observed during the mid-log phase. Following a curing procedure with novobiocin, only 90-1330 lost the ability to produce BLIS, suggesting that the BLIS might be plasmid encoded. Analysis of the inhibitory spectra revealed that the BLIS-producing strains also inhibited the growth of Actinobacillus minor, Actinobacillus porcinus, Enterococcus durans, Micrococcus luteus, Streptococcus agalactiae, Streptococcus dysgalactiae subsp. dysgalactiae, Streptococcus equi subsp. zooepidemicus, and S. dysgalactiae subsp. equisimilis. This study reports for the first time the ability of the swine pathogen S. suis serotype 2 to produce BLIS with the characteristics of classic bacteriocins. Further studies are required to investigate the possibility of using bacteriocin-producing strains to prevent swine infections caused by virulent strains of S. suis serotype 2.

Animals↗

PubMed2002

Dissemination of the phage-associated novel superantigen gene speL in recent invasive and noninvasive Streptococcus pyogenes M3/T3 isolates in Japan.

In Japan, more than 10% of streptococcal toxic shock-like syndrome (TSLS) cases have been caused by Streptococcus pyogenes M3/T3 isolates since the first reported TSLS case in 1992. Most M3/T3 isolates from TSLS or severe invasive infection cases during 1992 to 2001 and those from noninvasive cases during this period are indistinguishable in pulsed-field gel electropherograms. The longest fragments of these recent isolates were 300 kb in size, whereas those of isolates recovered during or before 1973 were 260 kb in size. These 260- and 300-kb fragments hybridized to each other, suggesting the acquisition of an about 40-kb fragment by the recent isolates. The whole part of the acquired fragment was cloned from the first Japanese TSLS isolate, NIH1, and its nucleotide sequence was determined. The 41,796-bp fragment is temperate phage phiNIH1.1, containing a new superantigen gene speL near its right attachment site. The C-terminal part of the deduced amino acid sequence of speL has 48 and 46% similarity with well-characterized erythrogenic toxin SpeC and the most potent superantigen, SmeZ-2, respectively. None of 10 T3 isolates recovered during or before 1973 has speL, whereas all of 18 M3/T3 isolates recovered during or after 1992 and, surprisingly, Streptococcus equi subsp. equi ATCC 9527 do have this gene. Though plaques could not be obtained from phiNIH1.1, its DNA became detectable from the phage particle fraction upon mitomycin C induction, showing that this phage is not defective. A horizontal transfer of the phage carrying speL may explain the observed change in M3/T3 S. pyogenes isolates in Japan.

Amino Acid Sequence↗

PubMed1996

M proteins of group C streptococci isolated from patients with acute pharyngitis.

We studied 15 strains of group C (Streptococcus equi subsp. equisimilis) [corrected] isolated from the throats of college students with acute pharyngitis and 5 strains isolated from patients with noninfectious problems. Nineteen of the 20 strains resisted phagocytic killing during incubation in normal human blood, suggesting that they might express M proteins. Genomic DNA from all 20 strains hybridized with a probe corresponding to the carboxyterminal one-third of the group A M-protein gene emm24, a region that is highly conserved among M proteins of group A and group G streptococci. The DNA sequences of the N-terminal (variable) regions of the M-protein-encoding genes from two disease-associated group C isolates and one control isolate were determined. The predicted amino acid sequences of the two pharyngitis strains were identical and were 88% homologous to the amino acid sequence of a group G M-protein gene. The predicted terminal amino acid sequence of the control strain does not correspond to any such sequences in the GenBank database. All three strains studied possess the conserved region domain common to class I group A M-protein types epidemiologically associated with rheumatic fever. These studies demonstrate the presence of M proteins in strains of S. equi subsp. equisimilis [corrected] isolated in cases of endemically occurring acute pharyngitis. Certain of these proteins are similar to those of group G streptococci, while others may represent new M types. The similarity in structure and function between M proteins of nonrheumatogenic serogroups and those of rheumatogenic group A streptococci suggests that factors other than or in addition to M protein per se are likely involved in the pathogenesis of rheumatic fever.

Acute Disease↗

PubMed1981

Serum levels of amoxycillin following its oral administration to thoroughbred foals.

Amoxycillin trihydrate was administered orally to 6 foals at dose rates of 13 mg/kg (low), 20 mg/kg (medium) and 30 mg/kg (high) and serum concentrations determined at intervals up to 8 h. Therapeutic serum levels of 1 microgram/ml persisted for 268 mins at a dose rate of 13 mg/kg, for 339 mins at 20 mg/kg and for 381 mins at 30 mg/kg. A 2 micrograms/ml serum level persisted for 198 mins at a dose rate of 13 mg/kg, for 268 mins at 20 mg/kg and for 311 mins at 30 mg/kg. To determine the spectra of its antibacterial activity, the minimum inhibitory concentrations of amoxycillin against 8 genera of common pathogenic bacteria in foals were examined. Of the bacteria tested, only Streptococcus equi, Strep zooepidemicus and Staphylococcus aureus (non-beta lactamase producing) would have been treated effectively with the serum levels which persisted in these foals. It is concluded that amoxycillin cannot be considered for broad spectrum use against pathogens likely to be encountered in the foal, but, because of its ease of administration orally, has a useful role in treating streptococcal infections and staphylococcal infections not caused by beta lactamase producers. If sensitivity testing of isolates is made, the drug may be useful also for treatment of some infections caused by the Gram-negative organisms Actinobacillus equuli and Salmonella typhimurium.

Administration, Oral↗

PubMed1975

[Immunoelectronmicroscopic localization of cell wall antigens in streptococci. II. Localization of the group-specific polysaccharide of group C streptococci with ferritin- and peroxidase-labelled Helix pomatia-agglutinin (author's transl)].

The carbohydrate of the cell wall of group C streptococci is one of the best known group-specific streptococcal antigens with respect to its chemical structure. In the present paper, the ultrastructural location of the carbohydrate was investigated by means of immunoelectronmicroscopic techniques. Besides group-specific antibodies, the specific binding of an agglutinin (protectin Anti-AHP) of the edible snail (Helix pomatia) to structures with terminal N-acetylgalactosamine was used to demonstrate the antigen. The agglutinin was extracted from the albumen gland of the snail and purified by column chromatography. By means of glutardialdehyde it was coupled with ferritin or horesradish peroxidase. The investigations were done on strains of Streptococcus equisimilis and Streptococcus equi. Str. pyogenes (group A streptococci) was used as a control. On whole cells of group C streptococci the carbohydrate was demonstrated on the surface of the triple-layered cell wall. Near the cross-wall the carbohydrate seems to be more concentrated. In the presence of N-acety-D-galactosamine the labelling of the cell wall was inhibited. N-acetyl-D-glucosamine did not have such an effect. On isolated cell walls, both the outer and the inner surface were tagged. This suggests that the group-specific carbohydrate covers the peptidoglycan (mucopeptide) on both sides. The cytoplasmic membrane shows no reaction.

Agglutinins↗

PubMed1989

Rapid detection of group C streptococci from animals by latex agglutination.

A group C latex agglutination reagent, included as the negative control in the PathoDx Strep A latex agglutination test (Diagnostic Products Corp., Los Angeles, Calif), was compared with culture for rapid detection of group C streptococci (Streptococcus equi, S. equisimilis, S. zooepidemicus, and S. dysgalactiae) from swabs of veterinary specimens. The overall sensitivity of the latex test was 78%, and specificity was 97.6%. Only 25% of S. dysgalactiae isolates were detected, thereby accounting for the relatively low sensitivity. Ninety-three percent of the group C streptococci other than S. dysgalactiae were isolated from horses. When the latex test was evaluated for detection of group C streptococci other than S. dysgalactiae from horses, the sensitivity and specificity were 95.3 and 100%, respectively. We found the group C latex agglutination test to be a rapid and accurate method for the detection of the major pathogenic group C streptococci from swabs of equine specimens.

Animals↗

PubMed1995

Isolation of a spermatozoa motility inhibiting factor from chicken seminal plasma with antibacterial property.

A 78-kDa spermatozoa motility inhibiting factor (SMIF) was purified from chicken (Gallus domesticus) seminal plasma by anion exchange (DE-53) followed by affinity chromatography on concanavalin A-Sepharose. The factor is thermostable and inhibited the spermatozoa motility in a dose dependent manner. In addition, SMIF inhibited the growth of gram negative bacteria, Pasteurella multocida but not gram positive Streptococcus equi. The factor lost its spermatozoa immobilizing property after treatment with trypsin, chymotrypsin or pepsin. The inhibition of SMIF by beta-mercaptoethanol suggest the involvement of disulfide bonds in its activity. Similarly, this property was lost in presence of chicken seminal plasma or incubating SMIF with anti-SMIF antibodies. Evidence is provided for the presence of a high molecular weight protein (> 100 kDa) in chicken seminal plasma that neutralizes the motility inhibiting property of SMIF. No significant decrease in spermatozoa ATP was observed in presence of SMIF suggesting that the loss of spermatozoa motility was due to factors other than depletion in cell's energy. Using anti-SMIF antibodies, a cross-reactive protein was identified in the blood, liver and reproductive tissues of chicken and the seminal plasma of cattle and buffalo. However, the cross-reactive protein failed to inhibit chicken spermatozoa motility. The significance of SMIF in chicken seminal plasma is discussed.

Animals↗

PubMed1996

Identification and molecular characterization of serological group C streptococci isolated from diseased pigs and monkeys in Indonesia.

The present study was designed to comparatively investigate 34 beta-hemolytic streptococci isolated from infected pigs and monkeys from various islands in Indonesia. According to the serological and biochemical data, all 34 isolates were Lancefield's serological group C streptococci and could be identified as Streptococcus equi subsp. zooepidemicus. Of the 34 group C streptococci investigated, 28 grew on solid media in large, mucoid colonies, in fluid media at a uniform turbidity, and in soft agar in diffuse colonies. A decapsulation test with a hyaluronidase-producing Staphylococcus aureus strain revealed the hyaluronic acid nature of the capsular material. The remaining six streptococci grew on solid media in small, nonmucoid colonies, in fluid media as sediment with clear supernatant, and in soft agar in compact colonies. Determination of surface hydrophobicity by salt aggregation revealed a hydrophilic surface for the encapsulated bacteria and a hydrophobic surface for the unencapsulated group C streptococci. To further analyze the epidemiological relationships, all 34 mucoid and nonmucoid isolates from pigs and monkeys were subjected to protein and DNA fingerprinting. The latter was performed by pulsed-field gel electrophoresis. The protein profiles of all 34 isolates and the DNA profiles of 32 isolates appeared to be identical, with the DNA profiles of 2 isolates being closely related, indicating that a single virulent clone is responsible for this disease outbreak in Indonesia.

Animals↗

PubMed1999

Cloning, expression, sequence analysis, and characterization of streptokinases secreted by porcine and equine isolates of Streptococcus equisimilis.

Streptokinases secreted by nonhuman isolates of group C streptococci (Streptococcus equi, S. equisimilis, and S. zooepidemicus) have been shown to bind to different mammalian plasminogens but exhibit preferential plasminogen activity. The streptokinase genes from S. equisimilis strains which activated either equine or porcine plasminogen were cloned, sequenced, and expressed in Escherichia coli. The streptokinase secreted by the equine isolate had little similarity to any known streptokinases secreted by either human or porcine isolates. The streptokinase secreted by the porcine isolate had limited structural and functional similarities to streptokinases secreted by human isolates. Plasminogen activation studies with immobilized (His)(6)-tagged recombinant streptokinases indicated that these recombinant streptokinases interacted with plasminogen in a manner similar to that observed when streptokinase and plasminogen interact in the fluid phase. Analysis of the cleavage products of the streptokinase-plasminogen interaction indicated that human, equine, and porcine plasminogens were all cleaved at the same highly conserved site. The site at which streptokinase was cleaved to form altered streptokinase (Sk*) was also determined. This study confirmed not only the presence of streptokinases in nonhuman S. equisimilis isolates but also that these proteins belong to a family of plasminogen activators more diverse than previously thought.

Amino Acid Sequence↗

PubMed2005

4-methylumbelliferone, a hyaluronan synthase suppressor, enhances the anticancer activity of gemcitabine in human pancreatic cancer cells.

Hyaluronan (HA) is a ubiquitous, major component of the pericellular matrix and is necessary for various physiological processes. It plays a very important role in biological barriers. We previously reported that 4-methylumbelliferone (MU) inhibits HA synthesis and pericellular HA matrix formation in cultured human skin fibroblasts, Streptococcus equi FM100, and B16F10 melanoma cells. We hypothesized that MU-mediated inhibition of HA synthesis and pericellular HA matrix formation would increase the efficacy of anticancer drugs. We have already demonstrated in vitro, using a sandwich binding protein assay and a particle exclusion assay, that MU inhibits HA synthesis and formation of the pericellular HA matrix, respectively, in human KP1-NL pancreatic cancer cells. AlamarBlue assay revealed that the anticancer effect of gemcitabine in KP1-NL cells was increased by pretreatment with MU. In vivo simultaneous administration of MU and gemcitabine to tumor-bearing mice with severe combined immunodeficiency disease (SCID) decreased the size of the primary and metastatic tumors more than did gemcitabine alone. These data strongly suggest that a combination of MU and gemcitabine is effective against human pancreatic cancer cells. MU may have potential as a chemosensitizer and may provide us with a new anticancer strategy.

Animals↗

PubMed1993

Eosinophilic synovitis following the intra-articular injection of bacterial antigen in horses.

Purified streptococcal M protein was injected into one intercarpal joint in three horses hyperimmunised with Streptococcus equi M protein vaccine. The contralateral joints were injected with pH adjusted polyionic solution. All antigen-injected joints developed a severe suppurative synovitis (mean synovial fluid nucleated cell count = 102,200 x 10(6) cells litre-1). Eosinophils were found in the synovial fluid and in synovial membrane biopsy specimens of two of the horses. Immune complexes were not demonstrated in the synovial membrane. Two horses are described that developed synovial fluid eosinophilia after the intra-articular injection of streptococcal antigen.

Animals↗

PubMed2005

Treatment strategies for mares with placentitis.

Equine placentitis, and resultant preterm labor, are important sources of fetal and neonatal loss. The primary cause of equine placentitis is infection of the placenta with Streptococcus equi subspecies zooepidemicus, which ascends through the caudal reproductive tract. Current treatment protocols for mares affected with placentitis are empirical. This paper reviews treatment approaches for resolving placentitis and preterm labor in both equine and non-equine species. Specific therapies reviewed include antimicrobial, anti-inflammatory, tocolytic, and progestin agents.

Animals↗

PubMed2006

Disease transmission in horses.

Bacterial, viral and protozoal infections may cause severe reproductive losses. The present paper reviews the risk factors, clinical signs and preventive measures for the most important venereal or potential sexually transmitted diseases in horses. The stallion and use of semen for artificial insemination represent major risk factors for the transmission of bacterial contaminants of the penis, including Streptococcus equi subspecies zooepidemicus, Pseudomonas aeruginosa and Klebsiella pneumoniae, known to cause endometritis and infertility in the mare. The role of the stallion in disease transmission is also due to the non-clinical manifestation of diseases such as contagious equine metritis and equine viral arteritis. Dourine has been eradicated from many countries, but continues to be a problem in other areas of the globe. Strategies for the prevention of introduction and transmission of diseases in breeding operation are discussed.

Animals↗

PubMed2006

Transrectal ultrasonography and plasma progestin profiles identifies feto-placental compromise in mares with experimentally induced placentitis.

Transrectal ultrasonography of the caudal uterus and a progestin profile were evaluated for accuracy in identifying mares with feto-placental compromise in a model of placentitis. Twenty-two pregnant ponies were divided into four groups: (1) control mares (n=5); (2) instrumented controls (n=2); (3) instrumented inoculated mares (n=11); (4) inoculated mares (n=4). Mares in Groups 3 and 4 were inoculated with Streptococcus equi subsp. zooepidemicus. Maternal plasma progestins, vulvar discharge, mammary gland development, combined thickness of the uterus and placenta (CTUP) and placental separation were evaluated weekly before instrumentation, inoculation or Day 320 (Groups 1 and 2) and, thereafter, either daily (first three measurements) or several times weekly (last two measurements). Plasma progestin profiles were plotted to identify pattern characteristics. An abbreviated profile was created, consisting of four progestin samples collected at 48-h intervals, with Sample 1 collected the day before inoculation or on Day 285 in controls. Profiles were considered abnormal if Samples 2, 3, or 4 increased or decreased by more than 50% of Sample 1. A CTUP>1.0 cm or placental separation were considered abnormal. Placentitis was confirmed by histology of fetal membranes. Control mares had normal progestin profiles, transrectal ultrasonographic and clinical examinations. Control foals were born after Day 329; six were viable and one died after dystocia. All inoculated mares developed placentitis and foaled before Day 314. Thirteen of 15 foals were not viable. All inoculated mares had abnormal progestin profiles and 13 of the 15 were identified by the abbreviated progestin profile. Transrectal CTUP was affected by gestational age and increased after inoculation (P<0.05). Nine of 15 inoculated mares had a CTUP>1.0 cm by 5-day post-inoculation. By performing both tests, 20 of 22 mares were correctly identified with respect to pregnancy outcome. However, three inoculated mares exhibited minimal clinical signs and likely would not be examined in a clinical setting. These tests were diagnostic for identifying feto-placental compromise in the mare.

Animals↗

PubMed1993

Effects of high-molecular-weight hyaluronates on the functions of guinea pig polymorphonuclear leukocytes.

The effect of hyaluronate (HA) with a molecular weight of 1,900,000 (HA190) purified from products of Streptococcus equi on various functions of guinea pig peritoneal polymorphonuclear leukocytes (PMNs) was examined. HA190 effectively inhibited both the chemotactic activity of PMNs for a chemotactic peptide, formyl-Met-Leu-Phe, and the phagocytic activity of the cells for serum-opsonized zymosan (SOZ) in a dose-dependent manner. When the inhibitory activity of the HAs of different molecular weights, including HA190, HA of molecular weight 800,000 (HA80), and HA of molecular weight 300,000 (HA30), was assessed, the HA190 showed the strongest inhibitory activity and HA30 the lowest activity. In contrast, significant inhibition of the superoxide generation by PMNs upon stimulation with SOZ was not observed even in the presence of HA190 and HA80 at the highest concentration used (2.5 mg/mL). This finding indicated that the HAs studied did not prevent the interaction of PMNs with stimuli. From these data it is concluded that the binding of high-molecular-weight HAs to the HA receptors on PMNs may produce intracellular signals that are responsible for suppression of phagocytosis and chemotaxis but not for superoxide generation.

Animals↗

PubMed1993

Strangles.

The etiology, epizootiology, pathogenesis, and clinical presentation of strangles are described. Streptococcus equi, the causative organism, is highly host-adapted to Equidae and shows no antigenic variation. Protective immunity apparently is mediated by a combination of serum opsonic and nasopharyngeal mucosal humoral responses. Vaccines based on M protein or inactivated bacterial suspensions may reduce the clinical attack rate by 50%, a level of protection much lower than that produced during recovery from strangles.

Animals↗

PubMed2005

[Aspergillus fumigatus in nasopharyngeal cavity of horses].

Aspergillus fumigatus is a cosmopolitan opportunistic fungal associated to rhinopharyngitis, sinusitis and guttural pouches infection with nasal discharges. All them are similar with Strangle's sign, the infectious disease produced by Streptococcus equi spp. The aim of this work was to detect A. fumigatus in healthy horses living in boxes and field. 226 nasopharyngeal swabbing samples were obtained by mycological routine. A. fumigatus was isolated in 26 (11.5%) horses.

Animals↗

PubMedDate not supplied

Antibacterial properties of a silver chloride-coated nylon wound dressing.

OBJECTIVE: A silver chloride-coated nylon wound dressing (Ag-WD) was evaluated in vitro for antimicrobial activity against five common equine wound pathogens. STUDY DESIGN: Bacterial susceptibility study. SAMPLE POPULATION: Equine wound pathogens: Escherichia coli, Klebsiella pneumoniae, Pseudomonas aeruginosa, Streptococcus equi subspecies zooepidemicus, and Staphylococcus aureus. METHODS: An inoculum of each pathogen was incubated directly with Ag-WD and quantitated after 24 to 48 hours of incubation. To determine if bactericidal activity of Ag-WD was contact dependent, an inoculum of E. coli and Staphylococcus aureus was incubated separately from Ag-WD by a filter and quantitated after 18 hours of incubation. Inductively coupled plasma emission spectrometry (ICP) determined the silver concentration of Mueller-Hinton broth containing Ag-WD after 24 hours of incubation. To establish if the rate of bacterial killing by Ag-WD differed from a constant silver concentration, pathogens were exposed to a silver concentration of 6.45 microg/mL and quantitated after 18 hours. RESULTS: Direct exposure to Ag-WD significantly reduced bacterial numbers after 15 minutes for K. pneumoniae, 30 minutes for E. coli, 1 hour for P. aeruginosa, and 2 hours for S. equi subspecies zooepidemicus and Staphylococcus aureus. Indirect exposure to Ag-WD resulted in > or =99.9% and > or =90% kill of the inoculum doses of E. coli at 2 hours and Staphylococcus aureus at 18 hours, respectively. Incubation of the pathogens at the constant silver concentration resulted in bacterial killing rates similar to those obtained by incubation with Ag-WD. CONCLUSIONS: In vitro, equine pathogens are effectively killed when exposed to Ag-WD, and the rate of bacterial killing by Ag-WD is similar to a constant silver concentration of 6.45 microg/mL. CLINICAL RELEVANCE: The in vitro antimicrobial properties of this silver-coated nylon wound dressing are promising for future prevention of equine wound infections.

Animals↗