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Title Pathogenesis of Chlamydial Infections( 本文（FULLTEXT) ) Title Pathogenesis of Chlamydial Infections( 本文（FULLTEXT) ) Author(s) RAJESH, CHAHOTA Report No.(Doctoral Degree) 博士(獣医学) 甲第226号 Issue Date 2007-03-13 Type 博士論文 Version author URL http://hdl.handle.net/20.500.12099/21409 ※この資料の著作権は、各資料の著者・学協会・出版社等に帰属します。 Pathogenesis of Chlamydial Infections !"#$%&'()*+%,-./0 2006 The United Graduate School of Veterinary Sciences, Gifu University, (Gifu University) RAJESH CHAHOTA Pathogenesis of Chlamydial Infections !"#$%&'()*+%,-./0 RAJESH CHAHOTA CONTENTS PREFACE……………………………………………………………………… 1 PART I Molecular Epidemiology, Genetic Diversity, Phylogeny and Virulence Analysis of Chlamydophila psittaci CHAPTER I: Study of molecular epizootiology of Chlamydophila psittaci among captive and feral avian species on the basis of VD2 region of ompA gene Introduction……………………………………………………………… 7 Materials and Methods…………………………………………………... 9 Results…………………………………………………………………… 16 Discussion……………………………………………………………….. 31 Summary……………………………………………………………….... 35 CHAPTER II: Analysis of genetic diversity and molecular phylogeny of the Chlamydophila psittaci strains prevalent among avian fauna and those associated with human psittacosis Introduction……………………………………………………………… 36 Materials and Methods…………………………………………………... 38 Results…………………………………………………………………… 42 Discussion……………………………………………………………….. 55 Summary………………………………………………………………… 59 CHAPTER III: Examination of virulence patterns of the Chlamydophila psittaci strains predominantly associated with avian chlamydiosis and human psittacosis using BALB/c mice Introduction……………………………………………………………… 60 Materials and Methods…………………………………………………... 63 Results…………………………………………………………………… 66 Discussion……………………………………………………………….. 74 Summary………………………………………………………………… 76 i PART II Investigation of the Emergence of Chlamydial Infection in Animals and Human Disease Conditions CHAPTER IV: Involvement of multiple Chlamydia suis genotypes in porcine conjunctivitis in commercial farms of Japan Introduction……………………………………………………………… 79 Materials and Methods…………………………………………………... 80 Results…………………………………………………………………… 86 Discussion……………………………………………………………….. 92 Summary………………………………………………………………… 98 CHAPTER V: Molecular characterization of the Chlamydophila caviae strain OK135, isolated from human genital tract infection, by analysis of some structural and functional genes Introduction……………………………………………………………. 99 Materials and Methods………………………………………………… 100 Results…………………………………………………………………. 103 Discussion……………………………………………………………... 108 Summary………………………………………………………………. 110 CONCLUSION………………………………………………………………… 112 REFERENCES…………………………………………………………………. 116 ACKNOWLEDGMENTS……………………………………………………… 140 ii PREFACE Chlamydiae are the obligate intracellular bacterial pathogens of animals and human beings (185, 188). They replicate within the cytoplasm of host cells, forming characteristic intracellular inclusion bodies. The chlamydiae undergo a unique biphasic developmental cycle characterized by two morphologically distinct forms, the elementary body (EB) and the reticulate body (RB). EBs are small (0.2–0.3 µm in diameter) infectious forms that are either endocytosed or phagocytosed by host cells. The EB within an intracytoplasmic inclusion, transforms into the larger (0.5–1.6 µm in diameter) intracellular non-infectious, metabolically active, RB. The RB multiplies by binary fission and fills the inclusion, which expands in size. RBs re-condense back into EBs via intermediate bodies towards the end of the cycle (24–48 hr, depending on species and strains) and are then released from the host cell by lysis or exocytosis to initiate another cycle of infection (75, 119, 131, 216). The EB, in contrast to the RB, is structurally rigid, resulting from the formation of extensive disulphide linkages between various cysteine-rich proteins in or associated with the outer membrane. The chlamydial outer membrane consists of lipopolysaccharide (LPS), polymorphic outer membrane proteins (POMPs), peptidoglycan and cysteine-rich proteins like major outer membrane protein (MOMP), OmcA and OmcB. The most important of these is the MOMP, which makes up 60% of the weight of the outer membrane. MOMP is about 40- to 45-Kda in mass and contains serovar- and species-specific epitopes. MOMP acts as porins, channels to allow exchange of molecules including nutrients. The high rigidity results in EBs being resistant to both chemical and physical factors and therefore, adapted for prolonged extracellular survival, an important factor in terms of chlamydial pathogenesis and control or treatment of chlamydial infections (75, 176). Earlier all chlamydiae were placed in order Chlamydiales, family Chlamydiaceae and one genus Chlamydia that included 4 species: C. trachomatis, C. psittaci, C. 1 pneumoniae and C. pecorum (52, 64, 132). Following reclassification of the order Chlamydiales in 1999, the family Chlamydiacaeae is now divided into two genera, Chlamydia and Chlamydophila (43). The genus Chlamydia comprises of Chlamydia trachomatis, Chlamydia suis and Chlamydia muridarum species. The genus Chlamydophila contains the species Chlamydophila pneumoniae, Chlamydophila psittaci, Chlamydophila abortus, Chlamydophila pecorum, Chlamydophila felis and Chlamydophila caviae. Chlamydiae are responsible for a diverse range of diseases in birds and mammals including humans. Among human chlamydial infections, C. trachomatis causes blindness (trachoma), sexually transmitted disease and infertility. C. pneumoniae is a cause of acute respiratory infection, coronary heart disease and chronic inflammatory lung diseases viz. chronic obstructive pulmonary disease and asthma (71, 105, 177). Animal chlamydial infections are caused by many chlamydial species. In genus Chlamydia, C. suis is associated with either asymptomatic infections or cause conjunctivitis, enteritis, pneumonia and genital tract infections in domesticated and wild porcine (99, 165, 183). C. muridarum causes either no disease or asymptomatic pneumonitis among mice and hamsters (40, 48). In genus Chlamydophila, C. pecorum is associated with chronic conditions viz. pneumonia, polyarthritis, conjunctivitis, enteritis, encephalomyelitis, metritis and mastitis among ruminants, swine and marsupials (40, 52). C. felis is responsible for “feline chlamydiosis”, which is characterized by rhinitis and conjunctivitis in cats, particularly kittens (192). Studies conducted in Japan have shown C. felis to be more widespread in stray cats than in pet cats (26, 57, 222). C. abortus is a cause of abortion in sheep, goats and responsible for reproductive failure in bovine, equine and porcine (40, 113, 160). C. psittaci causes “avian chlamydiosis” among many avian species (reported 460 avian species) and can also infect mammalian species (8, 94, 210). Avian chlamydiosis is characterized by diarrhea, anorexia, respiratory distress, sinusitis, rhinitis 2 and conjunctivitis (8, 188). C. caviae is a host specific pathogen of guinea pigs and causes either self-limiting or asymptomatic inclusion conjunctivitis among young (4-8 weeks old) guinea pigs (63, 101, 135). C. pneumoniae is also reported from marsupials and equines (62, 218). Animal chlamydial infections also have public health hazard significance. Among all chlamydial species infecting animals, C. psittaci, C. abortus and C. felis are potential zoonotic pathogens (113). C. abortus can cause severe, life-threatening disease in pregnant women. Infection in women during pregnancy can result in spontaneous abortion or stillbirths, which are typically preceded by several days of acute influenza-like illness, as well as renal failure, hepatic dysfunction, disseminated intravascular coagulation, and possibly death (25, 87). Cases of C. felis infection associated with conjunctivitis and atypical pneumonia in humans have been reported over the years (19, 113), but it is only recently that compelling evidence of transmission from cats to humans has been provided by way of molecular classification techniques (74). In Japan serological evidence of human C. felis infection has been reported (222). The zoonotic infection by C. psittaci, is particularly a significant occupational health hazard to the workers in poultry industry, aviaries, zoos and also pet shop owners, farmers and veterinarians. Infection in humans can vary in severity but is usually associated with respiratory disease. But multiple organs may become infected resulting in endocarditis, myocarditis, hepatitis, encephalitis or meningitis and result in deaths (113, 181, 210). The financial losses resulting from this disease, particularly those incurred in the poultry and pet industries, combined with the fact that this is the most common animal chlamydiosis transmissible to man, highlights the economic importance and public health significance of avian chlamydiosis. The incidences of zoonotic infection by C. caviae, C. pecorum, C. suis and C. muridarum are unknown (113). Defining the distribution of natural chlamydial infections in animals and human and its relationship to pathogenesis provides the fundamental basis for management and control 3 of chlamydial diseases. The final outcome of chlamydial infection in animals and human depends on large numbers of host and organism specific factors (217). Genetic and biological differences among various chlamydial species/strains are well known. Different pathobiotypes of C. trachomatis strains have been recognized, which include trachoma, urogenital and lymphogranuloma venereum (LGV) biovars. These biovars varies in tissue tropism and host specific cellular response
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