The genome sequence of Streptococcus equi subspecies equi (S. equi)(www.sanger.ac.uk) contains pa... more The genome sequence of Streptococcus equi subspecies equi (S. equi)(www.sanger.ac.uk) contains partial phage sequences which may affect virulence, gene regulation and expression. The present study was undertaken to characterize P9, a temperate bacteriophage of S. equi [J.G. Spanier, J.F. Timoney, Bacteriophages of Streptococcus equi, J. Gen. Virol., 35: 369–375, 1977.] to gain insight into genes that contributed to formation of
An outbreak of acute, fatal, hemorrhagic pneumonia was observed in more than 1,000 mixed breed do... more An outbreak of acute, fatal, hemorrhagic pneumonia was observed in more than 1,000 mixed breed dogs in a single animal shelter. The Department of Anatomic Pathology at the University of California at Davis School of Veterinary Medicine performed necropsies on dogs that were found moribund in acute respiratory distress or found dead with evidence of nasal bleeding. All dogs had hemothorax and an acute, fibrinosuppurative pneumonia. Large numbers of gram-positive cocci were observed within the lungs of all dogs and within septic thromboemboli of remote organs in about 50% of cases. Bacterial cultures from the dogs and their environment revealed widespread beta-hemolytic Streptococus equi subspecies zooepidemicus (Lancefield Group C). Extensive diagnostic testing failed to reveal the consistent presence of copathogens in individual cases. The clinical, epidemiologic, molecular biologic, and pathologic data indicate that a single clone of S. zooepidemicus was the cause of an acutely fatal respiratory infection in these dogs.
Although serologic data indicate horses in N. America are exposed to a variety of leptospiral ser... more Although serologic data indicate horses in N. America are exposed to a variety of leptospiral serovars, abortion is almost always associated with Leptospira interrogans serovar Pomona type kennewicki. A variety of wildlife including raccoons, white tailed deer, striped skunks, opossums, and red and grey foxes have been shown to host serovar Pomona and have therefore been suspect as sources of infection for pregnant mares. The aim of the present study was to examine genetic diversity in serovar Pomona type kennewicki in wildlife and in aborting mares. Our approach utilized PCR that targeted tandem repeats at the VNTR - 4 locus and a 1235 bp 5'-sequence of the lk73.5 (sph2) and adjacent upstream sequence unique to serovar Pomona. All isolates/specimens of equine origin in 1992 and 2008 yielded amplicons of 1235 and 595 bp, whereas 14 isolates/specimens from wildlife yielding a 1235 bp amplicon characteristic of serovar Pomona produced amplicons of 1300, 550 bp (3), 1300 bp (10), or 595 bp (6) with the VNTR - 4 primer set. Wildlife therefore hosted at least three different genetic variants of type kennewicki including the genetic variant that predominated in aborting mares. The data are consistent with other studies indicating specific genetic variants of type kennewicki show a strong tendency to be associated with a specific host. Levels of antibody in wildlife sera reactive with rLk73.5, rLig130 and sonicate of type kennewicki were poorly correlated with PCR data, although rLk73.5 was superior to rLig130 in detection of antibody responses. PCR is therefore a more reliable tool for studies of wildlife reservoirs of Leptospira sp. than serologic surveillance that targets host induced proteins or LPS-rich sonicate.
Evasion of phagocytosis is an important virulence determinant of Streptococcus equi (S. equi subs... more Evasion of phagocytosis is an important virulence determinant of Streptococcus equi (S. equi subsp. equi), the cause of equine strangles and distinguishes it from the closely related but much less virulent S. zooepidemicus (S. equi subsp. zooepidemicus). We describe Se18.9, a novel H factor binding protein secreted by S. equi but not by S. zooepidemicus that reduces deposition of C3 on the bacterial surface and significantly reduces the bactericidal activity of equine neutrophils suspended in normal serum for both S. equi and S. zooepidemicus. Se18.9 is secreted abundantly by actively dividing cells and is also bound to the bacterial surface. Strong serum and mucosal antibody responses are elicited in S. equi infected horses. Although a gene identical to se18.9 was not detected in S. zooepidemicus, sequences encoding proteins of similar size with similar signal peptide sequences were found in 3 of 12 randomly selected strains. Since Se18.9 is unique to S. equi, and immunoreactive with convalescent sera and mucosal IgA, it has potential for immunodiagnosis and for study of mucosal antibody response to S. equi.
Two pyrogenic mitogens, SePE-H and SePE-I, were characterized in Streptococcus equi, the cause of... more Two pyrogenic mitogens, SePE-H and SePE-I, were characterized in Streptococcus equi, the cause of equine strangles. SePE-H and SePE-I have molecular masses of 27.5 and 29.5 kDa, respectively, and each is almost identical to its counterpart in Streptococcus pyogenes M1. Both genes are adjacent to a gene encoding a phage muramidase of 49.7 kDa and are located immediately downstream from a phage genomic sequence almost identical to a similar phage sequence in S. pyogenes M1. Strong mitogenic responses were elicited by both proteins from horse peripheral blood mononuclear cells. However, although both were pyrogenic for rabbits, only SePE-I was pyrogenic in ponies. Convalescent sera contained antibody to each mitogen and horses recovered from strangles or immunized with SePE-I were resistant to the pyrogenic effect of SePE-I. The immunogenicity of SePE-I suggests that it should be included in new generation strangles vaccines. In isolates of S. equi sepe-I and sepe-H were consistently present but they were absent from the closely related Streptococcus zooepidemicus, suggesting that phage mediated transfer was an important event in the formation of the clonal, more virulent, S. equi from its putative S. zooepidemicus ancestor.
European Journal of Clinical Microbiology & Infectious Diseases, 2011
The present study has been formulated in order to detect an immunoreactive protein whose identifi... more The present study has been formulated in order to detect an immunoreactive protein whose identification can play a major role in the early diagnosis of disease. The identified protein will be produced by recombinant methods and used for the recombinant protein based ELISA. A comparison was made between the developed method and the gold standard MAT test to evaluate the serodiagnosis potential of the protein. The protein profile, immunoblot and MALDI-TOF analysis was carried out to identify the immunoreactive protein. The immunoreactive protein identified was used to develop ELISA for the diagnosis of leptospirosis using patients' sera with various clinical manifestations. The immunoreactive protein was identified as Leptospira GroEL chaperonin of molecular weight 60 kDa. The theoretical/experimental molecular weights, pI were found to be 58.5/60 kDa and 5.41/6, respectively. The overall results of the recombinant GroEL-IgM ELISAs showed cumulative sensitivity, specificity, positive predictive value, and negative predictive values of 90.6%, 94.9%, 94.6%, and 91.0%, respectively. The performance of such ELISA appeared better than that of any other serological tests previously evaluated for the diagnosis of leptospirosis in India. Thus, a highly conserved and immunogenic outer exposed GroEL protein during infection clearly merits further use in the serodiagnosis of leptospirosis.
ABCC9 genetic polymorphisms are associated with increased risk for various human diseases includi... more ABCC9 genetic polymorphisms are associated with increased risk for various human diseases including hippocampal sclerosis of aging (HS-Aging). The main goals of this study were 1> to detect the ABCC9 variants and define the specific 3' untranslated region (3'UTR) for each variant in human brain, and 2> to determine whether a polymorphism (rs704180) associated with risk for HS-Aging pathology is also associated with variation in ABCC9 transcript expression and/or splicing. Rapid amplification of ABCC9 cDNA ends (3' RACE) provided evidence of novel 3'UTR portions of ABCC9 in human brain. In silico and experimental studies were performed focusing on the single nucleotide polymorphism, rs704180. Analyses from multiple databases, focusing on rs704180 only, indicated that this risk allele is a local expression quantitative trait locus (eQTL). Analyses of RNA from human brains showed increased ABCC9 transcript levels in individuals with the risk genotype, correspondin...
The genome sequence of Streptococcus equi subspecies equi (S. equi)(www.sanger.ac.uk) contains pa... more The genome sequence of Streptococcus equi subspecies equi (S. equi)(www.sanger.ac.uk) contains partial phage sequences which may affect virulence, gene regulation and expression. The present study was undertaken to characterize P9, a temperate bacteriophage of S. equi [J.G. Spanier, J.F. Timoney, Bacteriophages of Streptococcus equi, J. Gen. Virol., 35: 369–375, 1977.] to gain insight into genes that contributed to formation of
An outbreak of acute, fatal, hemorrhagic pneumonia was observed in more than 1,000 mixed breed do... more An outbreak of acute, fatal, hemorrhagic pneumonia was observed in more than 1,000 mixed breed dogs in a single animal shelter. The Department of Anatomic Pathology at the University of California at Davis School of Veterinary Medicine performed necropsies on dogs that were found moribund in acute respiratory distress or found dead with evidence of nasal bleeding. All dogs had hemothorax and an acute, fibrinosuppurative pneumonia. Large numbers of gram-positive cocci were observed within the lungs of all dogs and within septic thromboemboli of remote organs in about 50% of cases. Bacterial cultures from the dogs and their environment revealed widespread beta-hemolytic Streptococus equi subspecies zooepidemicus (Lancefield Group C). Extensive diagnostic testing failed to reveal the consistent presence of copathogens in individual cases. The clinical, epidemiologic, molecular biologic, and pathologic data indicate that a single clone of S. zooepidemicus was the cause of an acutely fatal respiratory infection in these dogs.
Although serologic data indicate horses in N. America are exposed to a variety of leptospiral ser... more Although serologic data indicate horses in N. America are exposed to a variety of leptospiral serovars, abortion is almost always associated with Leptospira interrogans serovar Pomona type kennewicki. A variety of wildlife including raccoons, white tailed deer, striped skunks, opossums, and red and grey foxes have been shown to host serovar Pomona and have therefore been suspect as sources of infection for pregnant mares. The aim of the present study was to examine genetic diversity in serovar Pomona type kennewicki in wildlife and in aborting mares. Our approach utilized PCR that targeted tandem repeats at the VNTR - 4 locus and a 1235 bp 5'-sequence of the lk73.5 (sph2) and adjacent upstream sequence unique to serovar Pomona. All isolates/specimens of equine origin in 1992 and 2008 yielded amplicons of 1235 and 595 bp, whereas 14 isolates/specimens from wildlife yielding a 1235 bp amplicon characteristic of serovar Pomona produced amplicons of 1300, 550 bp (3), 1300 bp (10), or 595 bp (6) with the VNTR - 4 primer set. Wildlife therefore hosted at least three different genetic variants of type kennewicki including the genetic variant that predominated in aborting mares. The data are consistent with other studies indicating specific genetic variants of type kennewicki show a strong tendency to be associated with a specific host. Levels of antibody in wildlife sera reactive with rLk73.5, rLig130 and sonicate of type kennewicki were poorly correlated with PCR data, although rLk73.5 was superior to rLig130 in detection of antibody responses. PCR is therefore a more reliable tool for studies of wildlife reservoirs of Leptospira sp. than serologic surveillance that targets host induced proteins or LPS-rich sonicate.
Evasion of phagocytosis is an important virulence determinant of Streptococcus equi (S. equi subs... more Evasion of phagocytosis is an important virulence determinant of Streptococcus equi (S. equi subsp. equi), the cause of equine strangles and distinguishes it from the closely related but much less virulent S. zooepidemicus (S. equi subsp. zooepidemicus). We describe Se18.9, a novel H factor binding protein secreted by S. equi but not by S. zooepidemicus that reduces deposition of C3 on the bacterial surface and significantly reduces the bactericidal activity of equine neutrophils suspended in normal serum for both S. equi and S. zooepidemicus. Se18.9 is secreted abundantly by actively dividing cells and is also bound to the bacterial surface. Strong serum and mucosal antibody responses are elicited in S. equi infected horses. Although a gene identical to se18.9 was not detected in S. zooepidemicus, sequences encoding proteins of similar size with similar signal peptide sequences were found in 3 of 12 randomly selected strains. Since Se18.9 is unique to S. equi, and immunoreactive with convalescent sera and mucosal IgA, it has potential for immunodiagnosis and for study of mucosal antibody response to S. equi.
Two pyrogenic mitogens, SePE-H and SePE-I, were characterized in Streptococcus equi, the cause of... more Two pyrogenic mitogens, SePE-H and SePE-I, were characterized in Streptococcus equi, the cause of equine strangles. SePE-H and SePE-I have molecular masses of 27.5 and 29.5 kDa, respectively, and each is almost identical to its counterpart in Streptococcus pyogenes M1. Both genes are adjacent to a gene encoding a phage muramidase of 49.7 kDa and are located immediately downstream from a phage genomic sequence almost identical to a similar phage sequence in S. pyogenes M1. Strong mitogenic responses were elicited by both proteins from horse peripheral blood mononuclear cells. However, although both were pyrogenic for rabbits, only SePE-I was pyrogenic in ponies. Convalescent sera contained antibody to each mitogen and horses recovered from strangles or immunized with SePE-I were resistant to the pyrogenic effect of SePE-I. The immunogenicity of SePE-I suggests that it should be included in new generation strangles vaccines. In isolates of S. equi sepe-I and sepe-H were consistently present but they were absent from the closely related Streptococcus zooepidemicus, suggesting that phage mediated transfer was an important event in the formation of the clonal, more virulent, S. equi from its putative S. zooepidemicus ancestor.
European Journal of Clinical Microbiology & Infectious Diseases, 2011
The present study has been formulated in order to detect an immunoreactive protein whose identifi... more The present study has been formulated in order to detect an immunoreactive protein whose identification can play a major role in the early diagnosis of disease. The identified protein will be produced by recombinant methods and used for the recombinant protein based ELISA. A comparison was made between the developed method and the gold standard MAT test to evaluate the serodiagnosis potential of the protein. The protein profile, immunoblot and MALDI-TOF analysis was carried out to identify the immunoreactive protein. The immunoreactive protein identified was used to develop ELISA for the diagnosis of leptospirosis using patients' sera with various clinical manifestations. The immunoreactive protein was identified as Leptospira GroEL chaperonin of molecular weight 60 kDa. The theoretical/experimental molecular weights, pI were found to be 58.5/60 kDa and 5.41/6, respectively. The overall results of the recombinant GroEL-IgM ELISAs showed cumulative sensitivity, specificity, positive predictive value, and negative predictive values of 90.6%, 94.9%, 94.6%, and 91.0%, respectively. The performance of such ELISA appeared better than that of any other serological tests previously evaluated for the diagnosis of leptospirosis in India. Thus, a highly conserved and immunogenic outer exposed GroEL protein during infection clearly merits further use in the serodiagnosis of leptospirosis.
ABCC9 genetic polymorphisms are associated with increased risk for various human diseases includi... more ABCC9 genetic polymorphisms are associated with increased risk for various human diseases including hippocampal sclerosis of aging (HS-Aging). The main goals of this study were 1> to detect the ABCC9 variants and define the specific 3' untranslated region (3'UTR) for each variant in human brain, and 2> to determine whether a polymorphism (rs704180) associated with risk for HS-Aging pathology is also associated with variation in ABCC9 transcript expression and/or splicing. Rapid amplification of ABCC9 cDNA ends (3' RACE) provided evidence of novel 3'UTR portions of ABCC9 in human brain. In silico and experimental studies were performed focusing on the single nucleotide polymorphism, rs704180. Analyses from multiple databases, focusing on rs704180 only, indicated that this risk allele is a local expression quantitative trait locus (eQTL). Analyses of RNA from human brains showed increased ABCC9 transcript levels in individuals with the risk genotype, correspondin...
Uploads
Papers by Sergey Artiushin