Brucellosis Notes PDF

 BRUCELLOSIS - Brucellaceae family > Genus Brucella - Gram NEGATIVE - rod / coccobacilli - NON MOTILE - NON SPORULATED - NON CAPSULATED - NO NATIVE PLASMIDS - FLAGELLA...

 BRUCELLOSIS - Brucellaceae family > Genus Brucella - Gram NEGATIVE - rod / coccobacilli - NON MOTILE - NON SPORULATED - NON CAPSULATED - NO NATIVE PLASMIDS - FLAGELLA - STRICT AEROBIC - OIE LIST B - ZOONOTIC - OBLIGATE INTRACELLULAR PATHOGEN - !!!!!! CAN PASS TRHOUGHT INTACT SKIN !!!!!! - Species - Brucella abortus - Biovar 1 to 9 - CATTLE BRUCELLOSIS => ++++ biovar 1 - smt SR BRUCELLOSIS - HORSE BRUCELLOSIS - Brucella melitensis - SR BRUCELLOSIS => +++ Biovar 1, 2, 3 - smt CATTLE BRUCELLOSIS - assympto - Brucella suis - smt HORSE BRUCELLOSIS - smt CATTLE BRUCELLOSIS - similar B. abortus - Brucella canis - CANINE BRUCELLOSIS - smt CATTLE BRUCELLOSIS - Brucella ovis - agglutinate ONLY ANTIBRUCELLA R SERA - smt CATTLE BRUCELLOSIS - CONTAGIOUS RAM EPIDIDYMITIS - Non zoonotic - Resistance - +++ env if moist and/or organic - up 4m conta fluid: milk, urine, water, damp soil… - survive FREEZING + THAWING - Susceptibility - heat - sunlight - standard disinfectants - hypochlorite solutions - phenols - iodophors - pasteurization milk - Bioch - Oxidase + - Catalase + - Nitrate reductase + - Urease + - Surface ATG - A: +++ B. abortus - M: +++ B. Melitensis - R & Z: only in R colonial phase - CLINICAL - If ABOTOGENIC => SECOND PART GESTATION - Diagnostic - 1) Staining 22  - partial acid/fast - Special methods - KOSLOWSKI - KOSTER’S - STAMP - 2) Culture - Brucella agar - Modified Farrell’s Medium - Serum Dextrose Agar - 25-42°C - 3) Direct ID of pathogen - Immunofuorescence - +++ - ID BRUCELLA Sp thx - omp2 - bcsp31 - 4) SEROLOGY: - 1) STAT = serum tube agglutination - detection IgM or IgG - titer > 1:160 = + - 2) BBAT = buffered brucella ATG test - ROSE BENGAL AGGLUTINATION TEST - fast - qualitative (+/-) => require confirmatory diag by Complement Fix test (CFT) or ELISA - reduced efficacy in chronic cases or endemic areas - + = agglutination - limitations: - PROZONE PHENOMENON: if HIGH AMOUNT ATB => can appear negative - if chronic evolution => ATB can disappear up reappearance patho => negative - 3) COMPETITIVE ELISA - one of most common serologic methods - +: can control large quantities of sample (others only 1 by 1) - 4) BRUCELLIN SKIN TEST = BURNET’s INTRADERMAL TEST : RUMINANTS - screening unvaccinated herds - delayed hypersensitivity reaction - + : induration > 6mm in 24h - !!! ONLY FOR CHRONIC - 5) CFT - confirmatory test - + : >= 20 ICFTU/mL - 6) AGAR GEL IMMUNODIFUSION - 7) SPECIFIC TESTS - B. abortus - MILK RING TEST - cheap - surveillance dairy herd - POSITIVE = BLUE RING at surface + rest milk unstained - NEGATIVE = whole blood is blue - 3 consecutive tests on 3d - B. canis - BRUCEROCAN diag kit - B. ovis - BROVSET TEST = type of CFT 23  Brucellosis in CATTLE one of the most common contagious +communicable zoonotic disease in cattle: high rates morbidity + lifetime sterility => NOTIFIABLE DISEASE Cattle: abortion in the last trimester + possible permanent sterility + by reduced milk yield + high temperature + articular disorders Humans: undulant fever, general malaise, arthritis - ETIOLOGY - Alpha proteobacteria class > Rhizobiales order > Brucellaceae family > - Brucella abortus biovar 1 +++ - other sp: - Brucella melitensis (no CLINICAL signs) - Brucella suis (similar B. abortus) - Brucella ovis - Brucella canis - CHRONIC DISEASE + ABORTIONS - Tropism: REPRO TRACT + ARTICULAR DISORDERS - Resistance - SURVIVAL - survive FREEZING + THAWING - up 4m in conta fluid : MILK, URINE, WATER, DAMP SOIL - infectivity - 100d WINTER - 30d SUMMER - not free living in environment - SUSCEPTIBILITY - heat - sunlight - standard disinfectants - hypochlorite sol - phenols - iodophors - PASTEURIZATION of milk - EPIDEMIO - DISTRIB - WORLDWIDE excp where eradicated, eg: Denmark, Finland, Norway, Sweden, Romania - MOST PREVALENT ZOONOSIS DISEASE OIE LIST B - present in Europ - Absent in Romania - importance: - ZOONOTIC - Economic losses - reduced milk prod due abortion - losses of calves - culling - reduced export possibilities - loss of lost labor - vet costs - corset of research + eradication programs - => up 18$/buffalo in India - Hosts/ Susceptible species - Cattle + Bison + Buffalo , ELKS, CAMELIDS - higher receptivity = PREGNANT COWS - pregnant uterus = TARGET of FIXATION + MULTIPLICATION - higher seroprevalence = OLDER ANIMALS - infection dep - age - physio cond - gender 24 - SOURCES - Cattle - FEMALE - HIGH CONCENTRATION IN PREGNANT UTERUS - ABORTED FOETUS (+++ LUNGS & RENNET) - FOETAL MEMBRANES + LIQUIDS - PLACENTA (LESIONED COTYLEDONS) - VAGINAL DISCHARGE (before abortion or premature birth) - LOCHIA POST PARTUM - MAMMARY GLAND + MILK - MALE - SEMINAL SECRETIONS - DOGS & CATS + Some WILD (++ RODENTs) - URINE - Contamination / TRANSMISSION - direct contact - with inf animals - with carriers - natural mating (male to female) - intact or lesioned skin - indirect contact - contact with infected discharges - contact with aborted materia (foetus, membranes) - contact with infected env: litter, manure, feed, manure… - !!! ARTIFICIAL INSEMINATION (male to female) - HUMANS - ingestion raw, underpasteurized milk or milk products - intact or lesioned skin - Entry route - skin, ++ limb extremities + udder - digestive tract => ingestion - mucosal membranes => conjunctiva + vaginal - Contagiosity - ++ contagious - enzoonotic - zoonotic potential (farmers, slaughterhouse workers, vets..) 25 - PATHOGENESIS - FEMALES - Entry -> regional LN : x° > Lymph > transported in BLOOD thx MACROPHAGES: INTERMITTENT BACTEREMIA > Liver + Spleen + Udder LN -> then end bacteremia but x° in organs rich MACROPHAGE MONONUCLEAR TISSUE: LATENT INF > either resolve @+7w or invade PREGNANT UTERUS due ERYTHRITOL CHEMOTAXIS prod foetal tissue (++ present from +5m gest) & INTERMITTENT SHEDDING in MILK > PLACENTITIS + foetus ABORTION - !! NO PERSISTENCE IN UDDER BEFORE PUBERTY - PERSISTENCE LIVER LN SPLEEN EVEN IF SERONEG IN CHRONIC CASES - MALES - +2-3w infection -> invasion TESTIS + ACCESSORY GLANDS => ACUTE or CHRONIC SUPPURATIVE + NECROTIC INFLAMMATION => INFECTION SPERM + INFERTILITY - BOTH MALES & FEMALES - affect JOINTS + TENDINOUS FASCIA + SEROUS BURSAE - CLINICAL - Incub: 10d up 8m - FEMALE REPRO TRACT DISORDERS - usually assympto - new outbreaks: - ABORTIONS btw 6th to 9th month + PLACENTAL RETENTION + ENDOMETRITIS - past abortion: Continuous / Intermittent vaginal DISCHARGE 1-2w - smt: no abortion but WEAK newborns (LOCHIA) - MALES REPRO TRACT DISORDERS - modif general condition - reduced libido - uni/bilateral scrotum swelling - inflammation seminal vesicles - evolution on 2 to 4w then improvement (BCM CHRONIC) - BOTH JOINT DISORDERS - ++ JOINTS - bursitis - tenosynovitis - arthritis - periarthritis - LESIONS - FEMALE - Hyperemia + Edema genital tract - Uterus: Catarrhal endometritis - Foetal membranes: THICK + EDEMATOUS + HYPEREMIA + Fibrinopurulent deposits, hemorrhagic/necrotic placentitis + hemorrhagic cotyledons - ABORTUS - fibrinous pleuritis - increased liver + subcapsular necrotic foci + capsular fibrin deposits - MALES - orchitis + periorchitis - epididymitis: large brucellosis granuloma or atrophy + sclerosis - large necrosis or large abscesses => open & form caverns - Articular LESIONS - bursitis - tenosynovitis - arthritis - periarthritis - spondylitis - Cardiac LESIONS - vegetative endocarditis - Granulomatous inflammatory LESIONS - Repro tract - Udder - LN & lymphoid tissue - Joints 26 - DIAGNOSIS - CLINICAL x necropsy x lab test - LAB TEST - 1) Detection bacteria/ agent - 1) Culture - low diag value (reduced quant bact) - only ACUTE STAGE - require SEVERAL TEST due TRANSIENT BACTEREMIA - Specific media - 2) IMMUNOFLUORESCENCE: abortion samples - 3) PCR - 2) Detection Immune resp - 1) STAT = Serum Tube Agglutination - 2) ROSE BENGAL AGGLUTINATION TEST - 3) CFT - 4) COMPETITIVE ELISA - 5) MILK RING TEST - 6) BRUCELLIN SKIN TEST = BURNET’S INTRADERMAL TEST - DD - Trichomoniasis - Vibrisosis - Listeriosis: + neuro signs - Infectious Bovine Rhinotracheitis: + respi & digest symptoms - Mycoses & mycotoxins - PREVENTION - general measures if not endemic - hygiene - trade of animals : serologic testing - serologic surveillance with rose bengal test or CFT - endemic - vaccine: B. abortus strains 19 & RB51 vaccines - S19: calves 6-9m + annual booster - RB51: heifers 4-10m + booster 12-16m - humans: THERAPY ONLY, no VACCINE - TTT - no efficient therapy for cattle bse brucella in RES + macrophages: ATB can’t reach - CONTROL - SURVEILLANCE + ID positive animals (RBT + CFT, ring milk test) + confirm PCR => CULLING - shall involve all workers on chain (farmers, slaughterhouse, butcher, consumers, vets - education workers - basic security equipment - avoid consumption RAW dairy products - carcasses can be used in hum consumption IF COOKED - Eradication measures - ID + SEGREGATE & CULLED - test semen from reproductor males 27  Brucellosis in SR contagious and communicable zoonotic disease in SR with high rates of morbidity and lifetime sterility => NOTIFIABLE DISEASE SR: abortion in the last trimester + possible permanent sterility + reduced milk yield + high temp + articular disorders Humans: undulant fever, general malaise, arthritis - ETIOLOGY - Brucella melitensis biovar 1, 2, 3 + RARE Brucella abortus (if close contact positive CATTLE) - 2 types - CHRONIC + ASSYMPTO EVOL - CHARACT SIMILAR CATTLE - abortion + placenta retention + metritis - orchitis - bursitis - one most serious zoonosis in sheeps, ++ contagious & enzootic - EPIDEMIOLOGY - ++ Mediterranean disease - Susceptibility - sheeps & goats - lambs resistant during 1st m - Sources - infected animals shed - milk - urine - feces - fetal membranes + liquides + uterine discharge - Transmission - direct + indirect - intact or lesioned skin - mucosa - genital - GOAT ONLY - DIGESTIVE TRACT due grooming - PATHOGENESIS - SIMILAR CATTLE - brucella granuloma - CLINICAL - abortion 3-5th months - hemorrhagic discharge +5-7d post partum - ACUTE PHASE UNLIKE BOVINE POSSIBLE with neuro signs - some sheeps; acute/chronic bronchitis - LESIONS - brucella granuloma: SPLEEN, LIVER, UTERUS, TESTIS - testicle sclerosis, infertility - lobar pneumonia - increased LN - DIAG - 1) Identification of ATG - PCR - semen, foetus aborted, fetal membrane, affected organs - target same genes CATTLE - 2) Culture - fresh sample from aborted foetus - specific media - specific staining - 3) SEROLOGY - 1) STAT = Serum Tube Agglutination - 2) ROSE BENGAL AGGLUTINATION TEST - 3) COMPETITIVE ELISA - 4) BRUCELLIN SKIN TEST = BURNET’S INTRADERMAL TEST 28 - DD - Leptospirosis (+neuro signs, hemorrhagic urine + liver & kidney failure) - Campylobacteriosis - Q fever - Listeriosis = sillage disease in sheeps (+ neuro signs + digest signs) - Salmonellosis - S. abortus ovis (+ digestive signs) - S. typhimurium in lambs (+ dysentery (hemorrhagic diarrhea)) - PREVENTION - ++ SURVEILLANCE (CFT, ROSE BENGAL TEST) - QUARANTINE MANDATORY for newly imported animals - SEROLOGIC SURVEILLANCE - all imported SR during quarantine by SERUM AGGLUT TEST - PROPHYLAXIS - immunization Rev1 = LIVE VACCINE, not reco during pregnancy - !!!! interfere with serology : no diff btw natural & vaccination immune resp - CONTROL - test & slaughter program - conta area: repeated disinfections + sanitary void 29  Brucellosis in HORSES = Suppurative bursitis = Fistulous Withers / Poll Evil rare equine pathology causing mainly suppurative bursitis: pus-filled inflammation of connective tissue over the shoulders or poll and occasionally abortion, infection of the sexual organs in stallions, and infertility - ETIOLOGY - Brucella abortus: Main - Brucella suis - EPIDEMIOHir - Susceptibility - very RARE IN HORSES - Entry route - lesion @ withers due unfitted saddle - digestive route - Sources - inf animals (acute, chronic, assympto) - horses but also CATTLE + SWINE - aborting animals - normal birthing animals: infected placenta - fistulization of SUPRASPINOUS BURSITIS = CHARACTERISTIC CLINICAL OUTCOME - intact or lesioned skin => RQ GLOVES - PATHOGENESIS - ingestion -> oral mucosa -> regional LN -> transient bacteremia -> REPRO TISSUES + MUSCULOSKELETAL SYSTEM - CLINICAL - 1) ASYMPTOMATIC FORM - 2) CHRONIC FORM - Fistulous wither = septic supraspinous bursitis - Poll evil = septic supra atlantal bursitis - smt affect general condition - mid up late term abortion possible, RARE - LESIONS - fistulization of wither exudative inflammation - supra atlantal / supraspinous bursitis - joints + tendons affected - SC abscesses, metritis, osteitis, osteomyelitis - DIAGNOSIS - 1) AGAR GEL IMMUNOFLUORESCENCE - 2) STAT = Serum Tube Agglutination - 3) ROSE BENGAL AGGLUTINATION TEST - 4) CFT - PREVENTION - general non spe measures (avoid contact horses, cattle, swine, SR - avoid wither LESIONS - parasitic control: ONCHOCERCA spp - CONTROL - isolation positive - TREATMENT - TTT of high value horses, otherwise: culling - systemic ATB - ATB with activity in acidic intracellular environments: doxycycline, rifampicin - long term ttt - debridement + disinf + reconstructive procedures 30  Brucellosis in DOGS Inf + contagious disease: abortions females + epididymitis & testis atrophy & infertility males, zoonotic - ETIOLOGY - Brucella canis - EPIDEMIO - worldwide, ++ china, pakistan, mexico, uk - susceptibility - all dogs - Sources - inf animals (++ URINE) - aborted materia - Semen - Transmission - CLOSE CONTACT - MATING (male to females) - Dynamic - EPIZOOTIC for Brucella canis - SPORADIC + SUBCLI if other species - PATHOGENESIS - primary bacteremia + smt 2ndy & tertiary bacteremia dep bacterial load - target: - mononuclear phagocytic system + RES (LIVER, SPLEEN, LN) - STEROID DEPENDANT REPRODUCTIVE TISSUES (like ERYTHRON) - prostate, testis, epididymis, gravid uterus, placenta - usually CHRONIC EVOLUTION - CLINICAL - incub: 1-3w - Brucella abortus + suis + melitensis => ASSYMPTO or REDUCED CLINICAL - !! POSITIVE SEROLOGY - Female - Brucella canis => ABORTIGENIC ++ 7-8w gestation (post midterm, like OTHER SPECIES) - ENDOMETRITIS, can lead to STERILITY - BROWN VAGINAL DISCHARGES - Male - SCROTUM SWELLING + EPIDIDYMIS HYPERTROPHY then sclerosis up infertility, - Balanoposthitis, orchitis, scrotal dermatitis (quite specific) - Articular - BURSITIS - purulent tenosynovitis ++ limbs, - lumbosacral discospondylitis - Rare - pyo granulomatous dermatitis - meningitis - ocular disorders (uveitis !! similar distemper) - LESIONS - Abortion and stillborns: - head edema, sc CT congestion, - petechiae + ecchymosis + renal hemorrhage - male: - necrotic + purulent epididymitis & orchiepididymitis - thickened epididymis duct, - DIAGNOSIS - 1) SEROLOGY - !! carrier animals can be SERONEG - 1) STAT = Serum Tube Agglutination - 2) CFT - 3) Agar Gel Immunodiffusion - 4) ROSE BENGAL AGGLUTINATION TEST - 5) BRUCEROCAN diag kit 31  - 2) BACTERIOLOGY - BAC ISOLATION from conta tissues - PROGNOSIS: BAD - PREVENTION - buy animals from free areas - SERO SURVEILLANCE - modified rapid serum agglutination test : B. canis ATG + ROSE BENGAL - quarantine imported - CONTROL - isolation positive - SERO control - REMOVAL positive from repro - ++ disinfection - TREATMENT - high value anim - STREPTOMYCIN 25mg/kg, 2xd, 6d 32  Contagious Ram Epididymitis = Ovine Brucellosis with Brucella ovis contagious disease that impairs rams’ fertility: epididymis, testicle + accessory sexual gland alterations => NOTIFIABLE DISEASE - ETIOLOGY - Brucella ovis - AGGLUTINATE ONLY ANTIBRUCELLA R SERA - = CHRONIC DISEASE of OVINE, charact by - EPIDIDYMITIS / ORCHIEPIDIDYMITIS in RAM - INAPPARENT or ABORTIVE/STILLBORN in EWES - EPIDEMIO - Susceptibility - ++ ameliorated breeds - ++ adults - ++ males - Sources - sick & carriers - semen: up 2-4 years post inf - in acute phase - feces - urine - milk - Transmission - DIRECT - genital MALE to FEMALE => MAIN: ++ outbreaks in mating season - FEMALE TO MALE: VECTOR SHEEP INF - INDIRECT - CLOSE CONTACT - WATER - FOOD - PATHOGENESIS - similar - MAIN TARGET = EPIDIDYMIS EPITHELIUM OF TAIL + INTERSTITIAL TISSUE OF SEMINAL VESICLE => formation of CYSTS & DEGEN of EPITH inducing BLOCKAGE of CHANNELs & SEMEN STASIS => GRANULOMAS + TESTIS ATROPHY up calcification - CLINICAL - Incub: 8 to 12 w - RAMS - 1) ACUTE PHASE - general state alteration - m. tremors - high fever (3to8d) - diffuse erythematous LESIONS in genital area - 2) CHRONIC PHASE - edema testicular bursae, ++ accum exudate in vagina => uni / bilateral deformation - SEVERE PAIN - REFUSAL TO WALK or WALK with LIMBS APPART - refusal to mate - +1-2w > ADHESIONS btw vagina - Adv stages => ABSCESSES, FISTULATION - => leak THICK + YELLOW + FETID + PURULENT EXUDATE - modif semen: NECROZOOSPERMIA or AZOOSPERMIA + YELLOW-greenish up RED-brown ejaculate, MODIF SPERMATOZOA - EWE - rare, asymptomatic - rarely abortion 1-3m gestation followed septic metritis & infertility - stillborn or congenital malformations newborns 33 - LESIONS - RAMS - @ start - SCROTAL + EPIDIDYMIS EDEMA - advanced stages - EPIDIDYMIS - SCLEROSIS - CYSTS - abscesses - granulomas - rupture epididymis - TESTIS - atrophy - sclerosis - granuloma - abscesses - EWE - suppurative necrotizing placentitis - vaginitis - metritis - Abortus - degen LESIONS + diffuse necrosis or necro foci liver - HISTOPATH = BRUCELLOSIS GRANULOMA - DIAGNOSIS - 1) MICROSCOPY - SEMINAL SMEARS => modif KOSTER METHOD + STAMP + GRAM - FNA: EPIDIDYMIS or other LESIONS - Immunofluorescence for ATG ID - 2) BACTERIO - Specific media - 3) SEROLOGY - 1) CFT (BROVSET test) + ROSE BENGAL TEST= early detection seropositif - 2) ELISA - 3) SERUM AGGLUT REACTIONS - !!!! cant differentiate NATURAL INF & VACCINATED - DD - Actinobacillus seminis - Brucella abortus & Brucella melitensis - Campylobacter fetus subsp foetus - Pseudomonas spp - Listeria monocytogenes - PROGNO - vital = favo - eco = severe - PREVENTION - intro only from brucella ovis free flocks - test semen from repro rams - CFT test rams 2w before & after mating season & purchased rams + ewe - avoid contact with other flocks - IMMUNIZATION - VACCINES - 1) B19 stain: live attenuated B. abortus - 2) Rev1 strain: B. melitensis - => !!!! INTERFERE WITH SEROLOGIC SURVEILLANCE - youngs: post weaning - adults 2-3m before mating season - some countries => NOT ALLOWED - CONTROL - CULLING POSITIVE - disinfection - STRICT CONTROL POST REPOPULATION 34