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GENUS BORDETELLA GENUS BORDETELLA Gram-, rod motile & non-motile oxidase + catalase + non-fermentative amino acid as source of energy peritrichous flagella non-spore forming commensals on m.m. of upper respy tract of animals Colonies of B. bronchiseptica. B. bronchiseptica. HABITAT B. bronchiseptica - parasite of ciliated epithelium of respiratory tract of mammals - inhabit in upper respiratory tract of susceptible animals such as pig dog cat rabbit horse guinea pig etc. B. avium - upper respiratory tract of infected fowl CLASSIFICATION 1. B. bronchiseptica animals 2. B. avium turkey coryza (rhinotracheitis) 3. B. pertussis man-pertussis (whooping cough) 4. B. parapertussis man-parapertussis TRANSMISSION B. bronchiseptica mammals - 1rycause, airborne carrier animal, shed the organisms is the important source of organism B. avium poultry - direct contact, by aerosal, water, litter (environment sources) B. bronchiseptica PATHOGENESIS Disease of ciliated epithelium in respiratory tract exhibit phase changes correlated with virulence identified by colonial appearance Virulence phase (phase I) filamentous hemagglutinin pertactin, fimbriae (allow attachment to the cilia) Avirulence form (phase IV) after repeated subculture different colonial morphology PATHOGENICITY a mild form of atrophic rhinitis and bronchopneumonia rhinitis, sinusitis, tracheitis, bronchitis domestic and wild animals are affected most prevalence in swine, guinea pig rare in man no report in ruminant B. bronchiseptica - the agent alone causes temporary turbinate atrophy by disturbing osteoblast physiology, transient and self limiting - combined infection with Pasteurella multocida causing atrophic rhinitis [AR] - AR affects pig at 3 wks to 7 months leading to turbinate destruction and 2ry complication young animal, non-immune adult, more susceptible infection in adult, mild, subclinical Virulence Factor : Virulence Factor B. bronchiseptica and B.avium Activity B.bron B.avium Filamentous haemagglutinin Bind to cilia* + - Pertactin Bind to cells* + + Fimbriae Attachment to cell* + + Adenylate-cyclase -hemolysin Interfere phagocytic cell function + - Tracheal cytotoxin Inhibit ciliary fn, kills ciliated cells, induce skin necrosis, impair osteogenesis, toxic for osteoblast + + Dermonecrotic toxin osteotoxin LPS Stimulate cytokine release + + + + + + Pathogenesis B. bronchiseptica Score 5 Atrophic rhinitis. Canine infectious tracheobronchitis Kennel cough most prevalent respiratory complexes B. bronchiseptica, canine parainfluenza virus, canine adenovirus etc. *coughing, retching, dog remain alert, non-febrile * tenacious mucoid to mucopurulent exudate * variable involvment of lungs and adjacent lympnode * self-limiting unless complicated by bronchopneumonia * mild upper respiratory tract infection in cat TRANSMISSION Direct contact or aerosal Clothing, feeding utensil Kennel, petshop, animal shelter Bronchopneumonia in dog Canine infectious tracheobronchitis(kennel cough) IMMUNITY local antibody (IgA) is believed to block B. bronchiseptica colonization in pigs (4days after PI) but unable to remove attached bacteria clearance require several weeks ISOLATION & IDENTIFICATION 1. Nasal swab from pig, tracheal swab or sinus from chicks. 2. Tracheal aspiration fluid 3. Blood agar & Mac Conkey agar biochemical test B. avium (urease –ve) B. bronchiseptica) (urease +ve) 4. Virulent isolate agglutinate red blood cells (ovine, bovine, guinea pig), identify B. avium 5. Inoculate organism into guinea pig caused death 6. Serological tests which have been developed are of limited value Bordetella avium [turkey coryza] rhinotracheitis in turkey and fowl - highly contagious upper respiratory tract disease - morbidity is high, mortality is low (< 5%) - recover 2 wks after infection or longer (6 wks) - young chicken usually affected - environment induce infection Bordetella avium Turkey coryza * produce hemagglutinin agglutinate quinea pig RBC and correlates with pathogenicity spread from direct contact by aerosols and from environmental sources * mucus accumulates in the nares with swelling in the submaxillary sinuses * beak-breathing excessive lacrimation * 2ry infection with E.coli Diagnosis * clinical sign & gross pathology * isolation & identification B. avium from sinus and tracheal exudates * virulent isolate agglutinate guinea-pig RBC * ELISA Bordetella spp. Host B. bronchiseptica Pigs AR dogs canine infectious tracheobronchitis kitten pneumonia horse respiratory infection rabbit avium B. parapertussis B. Lab. rodent turkeys lambs Disease conditon upper respiratory infection bronchopneumonia coryza pneumonia GENUS BRUCELLA GENUS Gram-rod non-motile BRUCELLA catalase+, oxidase+ (except B.ovis, B.neotomae) not produce acid from CHO non-spore forming aerobe (microaerophilic) parasite of man and animal (zoonosis) all pathgenic - both male & female reproductive organ - reticuloendothelial system CLASSIFICATION CLASSIFICATION B. abortus B. suis B. melitensis B. ovis B. canis B. neotomae cattle swine goat & sheep sheep & esp. ram dog desert wood rat Zoonotic infection Brucellosis in man Undulant fever TRANSMISSION m. m. of intestinal tract, genital tract, conjunctiva, skin abrasion, inhalation ingestion - most common transplacental transmission PATHOGENESIS - able to survive and multiply within cells of reticuloendothelial system and tissue associated - facultative intracellular bacteria - granulomatous inflammatory reaction virulence factor ability to invade and survive within host inhibition of fusion of phagosome-lysosome is GENUS BRUCELLA major mechanism of intracellular survival major outer membrane & LPS -smooth form lack outer membrane & LPS - rough form- less virulent LPS - enhance intracellular survival various stress proteins allow the organism to adapt within macrophage PATHOGENICITY point of entry (m.m.) intracellular of monocytes & macrophages bacteremic phase regional lymphnodes (proliferate or killed) lymph & blood stream reproductive organs & associated gland in sexually mature animals granulomatous, nodules, abscess Specific host [cattle, swine, goat, sheep] organisms preferential localized at reproductive organ of pregnant animals mammary gland, gravid uterus,* placenta* associated lymph nodes testes, genital organs, epididymis* growth factor erythritol, fetal fluid,* chorion* etc. non-specific host mammary gland reticulo-endothelial system bacteremia BRUCELLOSIS IN ANIMALS abortion results from: • interfere with fetal circulation due to placentitis • endotoxin • fetal stress because of inflammatory response in fetal tissues BRUCELLOSIS IN ANIMALS Brucella abortus (Bang’s disease) cattle, (sheep) abortion at 5-6 months or more (once) • weak calves • infection of mammary gland & associated lymphnodes persist per year shed organisms in milk intermittently • not apparent • orchitis, epididymitis • organisms in semen Brucella suis swine • more chronic in reproductive organs • long time bacteremia with or without local infection occassion in uterus, metritis intermittent • abortion at any time, stilbirth, • decrease litter size, weak piglets, • arthritis in piglets • abscess in tissue orchitis, sterility (permanent or temporary) Brucella melitensis goat & sheep • the same as in cattle, arthritis • sheep more resistant than goat • mastitis may occur in goat Brucella ovis sheep (high specificity) • rarely abortion, placentitis more susceptible than female • orchitis, impaired fertility, epididymitis Brucella canis dog [only host] • abortion around 50 days, infertility • rapidly spread infertiliy, orchitis, epididymitis Brucellosis in man B. melitensis, B. suis B. abortus, B. canis Orchitis caused by B. melitensis B. ovis orchitis Brucellosis in man B. melitensis, B. suis B. abortus, B. canis transmission contact with excretion of infected animal skin abrasion inhalation ingestion Brucellosis as zoonosis. Routes of infection. Brucellosis in humans (Undulant fever) * 1ry disease of the reticulo-endothelial system * raw milk and dairy product made with unpasteurized milk are important sources of infection * skin abrasion, inhalation, ingestion are the routes of infection * present as fluctuating pyrexia, fatique muscle, and joint pain * mild lymphadenopathy, splenomegaly, hepatomegaly * osteomyelitis is common complication * severe infections B. melitensis B. suis biotype1 and 2 * moderately severe B. abortus * mild B. canis ISOLATION enriched media with blood, liver B. abortus, B. ovis need 10% C02 Live animal - fluid samples, abscess materials - blood, milk, semen, vaginal exudate from abortion blood culture for B. canis (long bacteremia) Carcass - organs - tissue in macrophage system, supramammary, retropharyngeal, internal iliac, lumbar, mesenteric lymph nodes - liver, spleen, uterus, joint fluid Aborted fetus - placenta, fetal membrane, abomasal content IDENTIFICATION 1. Biochemical tests, oxidase+, urease+ growth in dyes at different concentration [Thionin, Basic Fuchsin] 2. Rose-Bengal plate test (screening, agglutination) confirmed by CFT or ELISA 3. Slide agglutination for biotypes using A M R monospecific sera 4. Inoculate of affected tissues in guinea pig. [in case of contaminated samples or few organisms] - pure isolated from guinea pig - high titre against Brucella IMMUNITY vaccine apply in calf and cattle delayed-type hypersensitivity against endotoxin Differentiation of Brucella species and their biotypes species B. melitensis B. abortus biotypes agglutination in sera A M R 1 - + - 2 + - - 3 + + - 1 + - - 2 + - - 3 + - - 4 - + - 5 - + - 6 + - - 7 + + - 9 - + - Differentiation of Brucella species and their biotypes(cont.) species biotypes agglutination in sera A M R 1 + - - 2 + - - 3 + - - 4 + + - B. canis - - + B. ovis - - + B. neotome + - - B. suis Colonies of B. abortus in brucella agar. thionin Basic fuchsin GENUS ALCALIGENES GENUS ALCALIGENES Gram non-spore forming rod (bacilli) peritrichous flagella motile & non-motile non-fermentative oxidase+ catalase + HABITAT saprophyte •intestine of vertebrates • milk product • other foods, sea • decompose Alcaligenes group • maybe part of the normal human flora • isolated from human respiratory tract, renal system • occasionally isolated from urine, blood spinal fluid, wound, and abscess Alcaligenes faecalis recover from animals except poultry is infrequent is not ordinarily considered pathogenic Alcaligenes xylosoxidans (Achromobacter xylosoxidans) has been isolated from many body sites very uncommon as a sole cause of infection bacteremia (teeth rocking with forcep), meningitis (after gunshot wound) ISOLATION MacCongey agar and Blood agar Urease -ve A. faecalis