Genus BRUCELLA Dr.P.Sreenivasulu Reddy MD., Professor of Microbiology NARAYANA MEDICAL COLLEGE Nellore-1 Andhra Pradesh INDIA Many Names of Brucellosis Human Disease Malta Fever Undulant Fever Mediterranean Fever Rock Fever of Gibraltar Gastric Fever Animal Disease Bang’s Disease Enzootic Abortion Epizootic Abortion Slinking of Calves Ram Epididymitis Contagious Abortion Sir William Burnett (1779-1861) •Physician General to the British Navy •Differentiated the various fevers affecting soldiers Jeffery Allen Marston •Contracted Malta fever. •Described his own case in great detail. Sir David Bruce (1855-1931) •British Army physician and microbiologist •Discovered Micrococcus melitensis (July 9, 1887) Bernhard Bang (1848-1932) •Danish physician and veterinarian •Discovered Bacterium abortus (1897) could infect cattle, horses, sheep and goats. Alice Evans, American bacteriologist Credited with linking the organisms Similar morphology and pathology between: Bang’s Bacterium abortus Bruce’s Micrococcus melitensis Nomenclature today credited to Sir David Bruce Brucella abortus and Brucella melitensis. Morphological Profile Brucella spp. are small, Gram-negative, non-motile, non-spore-forming, coccobacilli . Intracellular microbes causing chronic disease. Cultural characters Aerobe. Best medium is Trypticase Soya Agar with selective agents (Polymyxin,Cyclohexamide). Small,moist,translucent and glistening colonies after 3 or more days of incubation. Species Biovar/ Serovar Natural Host Human Pathogen B. abortus 1-6, 9 cattle yes B.melitensis 1-3 goats, sheep yes B. suis 1, 3 swine yes 2 hares yes 4 reindeer, caribou yes 5 rodents yes B. canis none Dogs yes B. ovis none Sheep no B. neotomae none Desert wood rat no marine mammals ? B. maris (?) Biochemical reactions Catalase and Oxidase positive. Nitrates are reduced. Rapidly positive for urease. Negative for IMViC reactions. Susceptibility Killed at 600 C in 10 minutes Pasteurization of milk . Survival is long in refrigerated milk, ice creams and cheese. Transmission to Humans Conjunctiva or broken skin on contact with infected tissues Blood, urine, vaginal discharges, aborted fetuses, placentas Ingestion Raw milk & unpasteurized dairy products (Cheese) Rarely through undercooked meat Transmission to Humans Inhalation of infectious aerosols Inoculation with vaccines B. abortus strain 19, RB-51 B. melitensis Rev-1 Conjunctival splashes, injection Pens, stables, slaughter houses Person-to-person transmission is rare Incubation varies 5 days to three months Transmission in Animals Ingestion / contact with infected tissues or body fluids Venereal Mucous membranes, injections Swine, sheep, goats, dogs Fomites Who is at Risk? Occupational Disease Cattle ranchers/dairy farmers Veterinarians Abattoir workers Meat inspectors Lab workers Hunters Travelers Consumers of unpasteurized dairy products Pathogenecity and Immunity Followed by Entry Ingested by PMN and not killed (super oxide dismutase ,LPS and nucleotide substance). Spread to local LN, multiply and spread to blood. Reaches to RES (Spleen,Liver,BM,LN). Resist phagocytosis (production of AMP/GMP and TNF Inhibits the phagolysosome fusion). Spread (CNS,Heart,Joints,Respiratory,Skin,GU system) Host response leads to granuloma formation and necrosis. B. abortus (Contagious/ Infectious abortion) Worldwide Some countries have eradicated it Notifiable disease in many countries abortions, arthritic joints. Fever of Unknown Origin (FUO) Main causes for spontaneous abortion in animals are erythritol, which can promote infections in the fetus and placenta and lack of anti-Brucella antibody in amniotic fluid. B. canis Poorly understood 1-19% prevalence in United States contact with aborted fetuses and semen. Rarely causes disease in humans. B. suis Biovars 1 and 3 Free United Kingdom, Canada Eradicated Worldwide problem. Holland, Denmark Low Incidence Middle East, North Africa Human Disease Can affect any organ or system All patients have a cyclical fever Headache, weakness, arthralgia, depression, weight loss, fatigue, liver dysfunction Human Disease 20-60% of cases Osteoarticular complications Arthritis, spondylitis, osteomyelitis Hepatomegaly may occur Gastrointestinal complications 2-20% of cases Genitourinary involvement Orchitis and epididymitis most common Human Disease Neurological Cardiovascular Endocarditis resulting in death Chronic brucellosis is hard to define Depression, mental fatigue Length, type and response to treatment variable Localized infection Blood donations of infected persons should not be accepted Human Disease Congenitally infected infants Low birth weight Failure to thrive Jaundice Hepatomegaly Splenomegaly Respiratory difficulty General signs of sepsis (fever, vomiting) Asymptomatic Chronic brucellosis: It is hard to define - Length, type and response to treatment variable The localized form: -Bones and joints. -Lumbar spondylodiscitis, sacroiliitis is typical. -Orchitis also frequent. Sequelae are highly variable - Granulomatous hepatitis, Arthritis, spondylitis, Anaemia, leukopenia,thrombocytopenia, Meningitis, uveitis, optic neuritis, - Endocarditis. Granuloma of Liver Diagnosis in Humans Isolation of organism Blood, bone marrow, other tissues Serum agglutination test Four-fold or greater rise in titer Samples 2 weeks apart Immunofluorescence Method Organisms in clinical specimens PCR Blood cultures in trypticase soya broth (Castaneda’s method): Biphasic medium. Sample inoculated bottle has to be tilted to flow the broth over solid medium and keep it upright position at 370C. The growth of brucellae is extremely slow. Bone marrow cultures are more sensitive than blood. Demonstration of IgM , IgG and Ig A ELISA is the best method. Standard tube agglutination test: Anti -O-polysaccharide of LPS. Killed strains of B. abortus as antigen. Not useful for B.canis. Modified Tube agglutination test: 2-mercaptoethanol is added to serum before testing which causes disruption of IgM and only IgG are detected. Problems with TA test: Prozone phenomenon, Presence of blocking or non-agglutinating antibodies. False positives with cholera, tularemia,yersinia or vaccination . Contd….. Castaneda strip test: Strip with colored brucella antigen. On addition of patients serum, if antibodies are present, prevents the flow of serum. Brucellin Skin test: Delayed hypersensitivity. Molecular techniques: PCR Radiology: Alterations in infected vertebrae; Pedro Pons sign ( erosion of antero-superior corner of lumbar vertebrae) and marked osteophytosis are suspecious of brucellic spondylitis. Diagnosis in Animals Culture of urine and Milk. Rapid latex agglutination test, Rose Bengal card test. (Rose Bengal + B.abortus + Sodium azide) Contd…. Milk Ring Test: Frequently used test. A drop of colored brucella antigen (B.abortus/B.melitensis with hematoxylin) is added to milk in a test tube, incubated in a water bath at 700C for 40-50 minutes. Positive: Blue ring at the top leaving the milk unstained. Negative: No ring. Milk remains uniformly blue. Treatment of Choice Combination therapy has the best efficacy Doxycycline for 6 weeks in combination with Streptomycin for 2-3 weeks or Rifampin for 6 weeks CNS cases to treat for 6-9 months. Same for endocarditis cases plus surgical replacement of valves. Prognosis May last days, months, or years Recovery is common Disability is often pronounced About 5% of treated cases relapse Failure to complete the treatment regimen Sequestered infection requiring surgical drainage Case-fatality rate: <2% ( untreated) Endocarditis caused by B. melitensis Prevention and Control Education about risk of transmission Wear proper attire if dealing with infected animals/ tissues Farmer, veterinarian, abattoir worker, butcher, consumer, hunter, public Gloves, masks, goggles Avoid consumption of raw dairy products Prevention and Control Immunize in areas of high prevalence Young goats and sheep with Rev-1 Calves with RB51 No human vaccine Eradicate reservoir Identify, segregate, and / or cull infected animals References: www.slideshare.net