BRUCELLOSIS

Bang's disease, Gibraltar fever, Malta fever, Maltese fever, Mediterranean fever, rock fever, or undulant fever.

Submitted to:- Dr.Rashmi Singh Submitted by:- vikas

Classification

• Phylum:- Proteobacteria• Class:- Alpha Proteobacteria• 1)Family:- Brucellaceae• Genus :- Brucella• Genus :- Mycoplana• Genus :- Orchrobactrum• 2)Family:- Bartonellaceae• Genus :- Bartonella

Species :- Species :- B.abortusB.abortus:-cattle:-cattle

B. melitensisB. melitensis:-goat, :-goat, B.suisB.suis:-pigs, :-pigs, B.canisB.canis:-:-dogs, coyotes, foxes.dogs, coyotes, foxes.

B.ovisB.ovis:-sheep, :-sheep, B.neotomaeB.neotomae:-rodent :-rodent (wood rat), (wood rat), B.marisB.maris:-marine mammals.:-marine mammals.

Humans:-Humans:-B.abortus, B.melitensis, B.suis, B.abortus, B.melitensis, B.suis, B.canis.B.canis.

HISTORY• The disease now called brucellosis,

under the name "Mediterranean fever", first came to the attention of British medical officers in Malta during the Crimean War in the 1850s. The causal relationship between organism and disease was first established by Dr. David Bruce in 1887.

In 1897, In 1897, Danish veterinarian Bernhard Bang isolated isolated Brucella abortus as the agent, and as the agent, and the additional name "Bang's disease" was the additional name "Bang's disease" was assigned. In modern usage, "Bang's assigned. In modern usage, "Bang's disease" is often shortened to just "disease" is often shortened to just "BangsBangs“.“.

Maltese doctor and archaeologist Sir doctor and archaeologist Sir Themistocles Zammit identified identified unpasteurized milk as the major source of unpasteurized milk as the major source of the pathogen in 1905, and it has since the pathogen in 1905, and it has since become known as Malta Fever, or become known as Malta Fever, or deni rqiqdeni rqiq locally.locally.

In cattle this disease is also known as In cattle this disease is also known as contagious abortioncontagious abortion and and infectious infectious abortionabortion..

• The popular name "undulant fever" originates from the characteristic undulance (or "wave-like" nature) of the fever which rises and falls over weeks in untreated patients. In the 20th century, this name, along with "brucellosis", gradually replaced the 19th century names "Mediterranean fever" and "Malta fever".

• In 1989, Saudi Arabian neurologists discovered neurobrucellosis, a neurological involvement in brucellosis.

The following obsolete names have previously been The following obsolete names have previously been applied to brucellosis:applied to brucellosis:

BrucelliasisBrucelliasis Bruce's septicemia Bruce's septicemia Continued fever Continued fever Crimean fever Crimean fever Cyprus fever Cyprus fever Febris melitensisFebris melitensis Febris undulansFebris undulans

Goat fever Goat fever Melitensis septicemiaMelitensis septicemia Melitococcosis Melitococcosis Milk sickness Milk sickness Mountain fever Mountain fever Neapolitan fever Neapolitan fever Slow feverSlow fever

Morphology, cultural & Morphology, cultural & biochemical characteristicbiochemical characteristic

Small ,gram negative,cocobacilli,0.6-1.5in Small ,gram negative,cocobacilli,0.6-1.5in length,0.5-0.7 micromtr. in breadth.length,0.5-0.7 micromtr. in breadth.

Non motile, non spore forming, Non motile, non spore forming, carboxyphillic(5-10% CO2), non acid fast. carboxyphillic(5-10% CO2), non acid fast.

Catalase+, urease+ (except Catalase+, urease+ (except B.ovisB.ovis), ), oxidase+ (except oxidase+ (except B.ovis B.ovis & & B.neotomae)B.neotomae), do , do not produce any acid from the not produce any acid from the carbohydrate.carbohydrate.

Cannot live apart from host.Cannot live apart from host. Pathogenic and facultative intracellular (in Pathogenic and facultative intracellular (in

mQs).mQs).

Resist decolourisation by dilute acids Resist decolourisation by dilute acids &alkalies so, MODIFIED ZIEHL NELSON’S &alkalies so, MODIFIED ZIEHL NELSON’S staining is done.staining is done.➩0.5% acetic acid used & ➩0.5% acetic acid used & organism observed as red cocobacilli.organism observed as red cocobacilli.

Stains red with Macchiavello stain.Stains red with Macchiavello stain. Media supplemented with blood or serum.Media supplemented with blood or serum. Organism appears after 2-3 days at 37 Organism appears after 2-3 days at 37

degrees in the presence of CO2.degrees in the presence of CO2. Selective media:- serum dextrose agar Selective media:- serum dextrose agar

media, liver dextrose agar media.media, liver dextrose agar media. After 3-5 days pinpoint, smooth, glistening, After 3-5 days pinpoint, smooth, glistening,

bluish, translucent colonies appear. As they bluish, translucent colonies appear. As they age the colonies appear opaque and about age the colonies appear opaque and about 2-3 mm in dia. 2-3 mm in dia.

Erythritol attracts the organism.Erythritol attracts the organism. Present in RE system of reproductive tract.Present in RE system of reproductive tract.

Cell structure & compositionCell structure & composition

Typical gram-ve cell wallTypical gram-ve cell wall Dominant surface Ag :- located on Dominant surface Ag :- located on

Lipopolysaccharide (A& M).Lipopolysaccharide (A& M). Peptidoglycan layer is quite prominent.Peptidoglycan layer is quite prominent. Periplasmic space varies 3-30nm.Periplasmic space varies 3-30nm. Cytoplasmic membrane is typical 3 layered Cytoplasmic membrane is typical 3 layered

lipoprotein membrane.lipoprotein membrane. Mol% G+C of DNA is 58-59.Mol% G+C of DNA is 58-59. 2 circular chromosomes. Plasmid DNA is –nt. 2 circular chromosomes. Plasmid DNA is –nt.

Cell productsCell products

1)Cell wall:-LPS an important virulence 1)Cell wall:-LPS an important virulence determinant.determinant.

Lipid A & length of the side chain in the O Lipid A & length of the side chain in the O repeat unit hinders attachment of MHC to the repeat unit hinders attachment of MHC to the outer membrane.outer membrane.

LPS binds to LPS binding proteinLPS binds to LPS binding protein➩ CD➩ CD14 14

➩➩CDCD1414--LPS complex binds to Toll like receptor LPS complex binds to Toll like receptor proteins on surface of mQs triggering release proteins on surface of mQs triggering release of proinflammatory cytokines.of proinflammatory cytokines.

Also LPS aids in survival of Also LPS aids in survival of Brucella Brucella in mQs.in mQs.

• 2) Outer membrane poteins:- porin proteins in outer membrane stimulate DTH.

• 3) Erythritol• 4) Miscellaneous products:-

Adenine & Guanine monophosphate.

PATHOGENESIS Depends on :- 1)Dose and no. of infective organism 2)Virulence 3) Host susceptibility rough forms are less virulent. Virulent forms phagocytosed ➩ regional L.N

➩ organism multiply in mQs ➩ thoracic duct ➩ blood stream ➩parenchymatous organs (mQs and parenchymal cells).

Interfere with phagosome and Interfere with phagosome and lysosome fussion and thus, survive in lysosome fussion and thus, survive in mQs and not in nuetrophils.mQs and not in nuetrophils.

Superoxide dismutase and catalase Superoxide dismutase and catalase help in resistance against oxidative help in resistance against oxidative killing & organism spreads to killing & organism spreads to reproductive tract.reproductive tract.

Intracellular multiplication Intracellular multiplication ➩ ➩ mononuclear infilteration ➩ giant cell mononuclear infilteration ➩ giant cell and granuloma formation.and granuloma formation.

Foci in lymphtic tissues, liver, spleen, Foci in lymphtic tissues, liver, spleen, bone marrow etc. ➩ abcess formation.bone marrow etc. ➩ abcess formation.

Antigenic natureAntigenic nature

Smooth and rough forms. Smooth and rough forms. Change from S to R forms due to:-Change from S to R forms due to:-

loss of virulence, tendency of loss of virulence, tendency of autoagglutination, and loss of Ag that autoagglutination, and loss of Ag that elicit aglutinins specific for smooth elicit aglutinins specific for smooth strains.strains.

B.abortusB.abortus, , B.melitensisB.melitensis, , B.suisB.suis:- S :- S forms.forms.

Possess two important surface Ags’ Possess two important surface Ags’ i.e A&M.i.e A&M.

Smooth & Rough formsSmooth & Rough forms

Smooth formSmooth form

1.1. Colonies are smooth, Colonies are smooth, glycening, bluish, & glycening, bluish, & translucent after 3-5 translucent after 3-5 days of incubation.days of incubation.

2.2. B.melitensisB.melitensis, , B.abortusB.abortus, , B.suisB.suis. .

Rough formRough form

1.1. Colonies are dull, Colonies are dull, yellowish, opaque, & yellowish, opaque, & friable in appearance.friable in appearance.

2.2. B.ovisB.ovis, , B.canisB.canis..

• B.abortusB.abortus :- more A. :- more A.

• B.melitensisB.melitensis :- more M :- more M

• B.suisB.suis :-intermediate pattern but :-intermediate pattern but more of A.more of A.

• B.canisB.canis & & B.ovisB.ovis has R surface Ag has R surface Ag but not M or A.but not M or A.

• B.abortusB.abortus share an Ag with some share an Ag with some strains of Yersinia enterocoliticus & strains of Yersinia enterocoliticus & so, a cross reaction develops in so, a cross reaction develops in serology. serology.

Brucellosis in cattleBrucellosis in cattle The bacterium The bacterium Brucella abortusBrucella abortus is the principal is the principal

cause of brucellosis in cattlecause of brucellosis in cattle.. The most common clinical signs of cattle The most common clinical signs of cattle

infected with infected with Brucella abortusBrucella abortus are high are high incidences of abortions(5-6incidences of abortions(5-6thth month), arthritic month), arthritic joints and retained joints and retained after-birth..

In India this disease is endemic & of great In India this disease is endemic & of great economic importance.economic importance.

Entry through ingestion ,infection of feces & Entry through ingestion ,infection of feces & urine of infected animal, penetration through urine of infected animal, penetration through skin & service by infected bull.skin & service by infected bull.

Incubation period is 30-60 days.Incubation period is 30-60 days.

• There are two main causes for spontaneous abortion in animals:-firstly due to erythritol, which can promote infections in the fetus and placenta.

• Second is due to the lack of anti-Brucella activity in the amniotic fluid.

• Males can also harbor the bacteria in their reproductive tracts, namely seminal vesicles, ampullae, testicles, and epididymides.

• In females:- gravid uterus in pregnant & in non pregnant in mammary glands.

• Sterility and infertility in infected herd.

Brucellosis in dogsBrucellosis in dogs

The causative agent of brucellosis in The causative agent of brucellosis in dogs is is Brucella canis..

It is transmitted to other dogs through It is transmitted to other dogs through breeding and contact with aborted breeding and contact with aborted fetuses. fetuses.

The bacteria in dogs normally infect The bacteria in dogs normally infect the genitals and the genitals and lymphatic system, , but can also spread to the eye, but can also spread to the eye, kidney, and intervertebral disc kidney, and intervertebral disc (causing discospondylitis). (causing discospondylitis).

Symptoms of brucellosis in Symptoms of brucellosis in dogs dogs

abortion in female dogs and scrotal abortion in female dogs and scrotal inflammation and orchitis (inflammation inflammation and orchitis (inflammation of the testicles) in males.of the testicles) in males.

Fever is uncommon.Fever is uncommon. Infection of the eye can cause uveitis, Infection of the eye can cause uveitis,

and infection of the intervertebral disc and infection of the intervertebral disc can cause pain or weakness. can cause pain or weakness.

Caprine and ovine brucellosis• B.melitensis• High abortion rates and orchitis

develops in males.• B.ovis causes ovine epididymitis in

lambs &placentitis in ewes. Sporadic abortions are reported. It is present in serum also.

• Test and slaughter policy done to control it.

Swine brucellosis

• B.suis

• Prolonged bacteremia and chronic inflammatory lesions in reproductive organs of sows and boars.

• Abortions ,still births & neonatal mortality.

• Secreted in semen.

• Modified live B.suis vaccine used in China.

Brucellosis in humansBrucellosis in humans

Brucellosis in humans is usually associated with:-Brucellosis in humans is usually associated with:-• the consumption of unpasteurized milk and the consumption of unpasteurized milk and

soft cheeses made from the milk of infected soft cheeses made from the milk of infected animals, primarily goats, infected with animals, primarily goats, infected with Brucella Brucella melitensis.melitensis.

• with occupational exposure of laboratory with occupational exposure of laboratory workers, veterinarians and slaughterhouse workers, veterinarians and slaughterhouse workers. workers.

• Some vaccines used in livestock, most notably Some vaccines used in livestock, most notably B. abortusB. abortus strain 19, also cause disease in strain 19, also cause disease in humans if accidentally injected. humans if accidentally injected.

The symptoms The symptoms Brucellosis induces inconstant fevers, Brucellosis induces inconstant fevers,

sweating, weakness, anaemia, headaches, sweating, weakness, anaemia, headaches, depression and muscular and bodily pain.depression and muscular and bodily pain.

Duration of disease is from few months to Duration of disease is from few months to years.years.

In the first stage of the disease, In the first stage of the disease, septicaemia occurs and leads to the septicaemia occurs and leads to the classic triad of undulant fevers, sweating classic triad of undulant fevers, sweating (often with characteristic smell, likened to (often with characteristic smell, likened to wet hay) and migratory arthralgia and wet hay) and migratory arthralgia and myalgia.myalgia.

If untreated, the disease can give origin to If untreated, the disease can give origin to focalizations or become chronic.focalizations or become chronic.

The focalizations of brucellosis occur usually in The focalizations of brucellosis occur usually in bones and joints and spondylodiscitis of lumbar bones and joints and spondylodiscitis of lumbar spine accompanied by sacroiliitis is very spine accompanied by sacroiliitis is very characteristic of this disease.characteristic of this disease.

Orchitis is also frequent in men.Orchitis is also frequent in men. Abortions not common in females due to Abortions not common in females due to

absence of erythritol.absence of erythritol. Raw milk is major source of contamination. Raw milk is major source of contamination.

Pasturization has reduced susceptibility of Pasturization has reduced susceptibility of brucella. brucella.

In blood tests, is characteristic the In blood tests, is characteristic the leukopenia and anaemia, some elevation leukopenia and anaemia, some elevation of AST and ALT and positivity of classic of AST and ALT and positivity of classic Bengal Rose is seen.Bengal Rose is seen.

This complex is, at least in Portugal, This complex is, at least in Portugal, known as the Malta fever. During episodes known as the Malta fever. During episodes of Malta fever, of Malta fever, melitococcemiamelitococcemia (presence (presence of brucellae in blood) can usually be of brucellae in blood) can usually be demonstrated by means of blood culture in demonstrated by means of blood culture in tryptose medium or Albini medium. tryptose medium or Albini medium.

Treatment

Resistant to B-lactam antibiotics. Treatment effective in humans if given early. Oral Doxycycline with Refampin for 30 days

is preferred to prolong treatment with a Tetracycline and Streptomycin.

Treatment of chronic disease is not always satisfactory ,coz is intracellular & there are inaccessible foci especially in bone.

Epidemiology

Humans acquire infection by handling infected tissue.

Common sources are abnormal fetuses, placentas ,post abortional uterine fluids, raw milk and milk products etc.

Persons at risk are veterinarians, slaughter house workers, ranchers, kennel workers, breeders etc.

In India ,disease is endemic in nature. 1943, prevalence of disease in goats ranged

b/w 10.2% &13.7%, Hissar. Low prevalence rate in goats & dairy animals

3 & 0.97% resp. from haldwani/bareilly region.

High PR of 22.5% of cattle, Orissa. Reported from >100 countries,230 thousand

cases annually around globe.

…

A serological survey of bovine brucellosis was performed in 23 states of India. A total of 30,437 bovine samples,comprising 23,284 cattle & 7,153 buffalo screened.

Overall prevalence,1.9% in cattle & 1.8% in buffalo.

Institute of Animal Health,Hebbal, Karnataka.

In India, infection from goat & sheep.

Geographical distribution B.abortus:-world wide common. viz, India,

Britain, Germany, Egypt, East Africa etc. B.melitensis:- New Zealand, Australia, North

America etc. B.suis:- Western & Central Europe, USA,

Argentina, Singapore, Arctic circle etc. B.ovis:-New Zealand, Australia, USA,

Romania, South Africa & South America, Czechoslovakia.

B.canis:- North America & parts of Europe. B.neotomae:- USA

IMMUNITYIMMUNITYCellular in nature.Cellular in nature.

Sensitized T lymphocytes release cytokines Sensitized T lymphocytes release cytokines that activate mQs ,which in turn control that activate mQs ,which in turn control BrucellaBrucella by reactive O2 intermediates. by reactive O2 intermediates. Brucella Brucella can activate NK cells to secrete IL-can activate NK cells to secrete IL-12.NK cells can then kill infected target cells.12.NK cells can then kill infected target cells.

More effective immunity develops when More effective immunity develops when animals are infected prior to sexual maturity.animals are infected prior to sexual maturity.

DiagnosisDiagnosis Samples:- blood, urine (males), fetal Samples:- blood, urine (males), fetal

organs and vaginal swabs.organs and vaginal swabs. +ive blood cultures:- reliable evidence +ive blood cultures:- reliable evidence

for B.canis.for B.canis.Other tests r as follows:-Other tests r as follows:- Rapid slide agglutination test. (RSAT)Rapid slide agglutination test. (RSAT) Agar gel immunodiffussion test Agar gel immunodiffussion test Tube agglutination testTube agglutination test ELISA, CFT, PCR, Hybridisation:- ELISA, CFT, PCR, Hybridisation:-

confirmatory tests.confirmatory tests. Indirect flurescent antibody test.Indirect flurescent antibody test. Brucellosis ring test/Bang ring test.Brucellosis ring test/Bang ring test.

MZN staining done in cotyledons, fetal MZN staining done in cotyledons, fetal discharges, & uterine discharges.discharges, & uterine discharges.

Intradermal Brucellin test can be done.Intradermal Brucellin test can be done. Agglutination test:- screening test for herd Agglutination test:- screening test for herd

for presence of reactors.for presence of reactors. SAT & TAT:- titre 1:40- anumal positive for SAT & TAT:- titre 1:40- anumal positive for

Brucella.Brucella. Rose Bengal Plate Test:- it identifies Rose Bengal Plate Test:- it identifies

subclass i.e IgG1.subclass i.e IgG1. Brucella milk ring test:- coloured Ag.Brucella milk ring test:- coloured Ag.

Vaccines Nonviable B.abortus 45/20. Attenuated live B.abortus strain 19 and RB51. A

single dose at 3-7 & 4-12 months of age is required with B.abortus strain19 & strain RB51 respectively. RB51 is comparatively a stable strain & is replacing strain 19 for calfhood vaccination.

Two doses 6 weeks apart in animals over 6 months of age are required with B.abortus45/20.

Strain 19 is occasionally shed in milk & can cause abortions in cattle.

RB51 rarely causes abortions. Bulls should not be vaccinated.

Orchitis develops in bulls.Orchitis develops in bulls. B.mellitensis B.mellitensis Rev1 :- attenuated vaccine : goats & Rev1 :- attenuated vaccine : goats &

sheep.sheep. Killed product i.e Killed product i.e B.mellitensisB.mellitensis H38 is also available. H38 is also available.

Conjunctival or sub cut. Kids & lambs vaccinated Conjunctival or sub cut. Kids & lambs vaccinated upto 6 months of age.upto 6 months of age.

Live vaccines are virulent for man.Live vaccines are virulent for man. Strain 19 vaccine discovered by Buck in 1930.Strain 19 vaccine discovered by Buck in 1930. Strain 45/20 is a rough strain & immune reaction to it Strain 45/20 is a rough strain & immune reaction to it

can be distinguished from that caused due to can be distinguished from that caused due to infection.infection.

Two dosses provide protection for an year. But can Two dosses provide protection for an year. But can get reverted to smooth forms and so, can be get reverted to smooth forms and so, can be dangerous.dangerous.

Outbreaks in IndiaOutbreaks in India Worldwide, reported incidence of human brucellosis in Worldwide, reported incidence of human brucellosis in

endemic disease areas varies widely,from <0.01 to >200 per endemic disease areas varies widely,from <0.01 to >200 per 100,000 population .100,000 population .

The true incidence of human brucellosis however, is The true incidence of human brucellosis however, is unknown for most countries and no data are available for unknown for most countries and no data are available for India. It has been estimated that the true incidence may be India. It has been estimated that the true incidence may be 25 times higher than the reported incidence dueto 25 times higher than the reported incidence dueto misdiagnosis and underreporting. misdiagnosis and underreporting.

Mostly it is seen that children are affected in large no. due Mostly it is seen that children are affected in large no. due to to the habit of consuming fresh goat milk and the close the habit of consuming fresh goat milk and the close contacts with animals .contacts with animals .

Fever and or joint pain notably of the knee and hip were Fever and or joint pain notably of the knee and hip were the only clinical manifestations in the majority of the the only clinical manifestations in the majority of the patientspatients . .

Since many patients with brucellosis present Since many patients with brucellosis present with fever as the only manifestation, other with fever as the only manifestation, other groups have investigated the prevalence of groups have investigated the prevalence of brucellosis in patients diagnosed with fever brucellosis in patients diagnosed with fever of unknown origin (FUO).of unknown origin (FUO).

In a study it was identified that 4 cases with In a study it was identified that 4 cases with acute brucellosis in a group of 121 patients acute brucellosis in a group of 121 patients with FUO.with FUO.

It was also identified that 28 (6.8%) It was also identified that 28 (6.8%) seropositiveseropositive

cases in a group of 414 patients with FUO cases in a group of 414 patients with FUO and 28 (0.8%) seropositive cases in a group and 28 (0.8%) seropositive cases in a group of 3,532 patients with FUO. The of 3,532 patients with FUO. The seroprevalence in specific risk groups such as seroprevalence in specific risk groups such as abattoir workers is much higher abattoir workers is much higher

•Thank you.