Babesiosis

Introduction
• Bovine babesiosis is a tick-borne parasitic disease t
hat results in significant morbidity and mortality in c
attle
• The economic losses can be considerable when ani
mals with no immunity are moved into an endemic
area
• Three species of Babesia cause most clinical c
ases in cattle:
• 1. Babesia bovis
• 2. B. bigemina are widespread in tropical and subt
ropical regions
• 3. B. divergens circulates in parts of Europe and p
ossibly in North Africa
• Most cattle Babesia do not seem to affect humans
• But B. divergens can cause rapidly progressing, life
threatening hemolytic anemia in people who hav
e had splenectomies.
 ETIOLOGY
• Family - Babesiidae
• Order - Piroplasmida
• Bovine babesiosis is usually caused by
• 1. Babesia bovis
• 2. B. bigemina
• 3. B. divergens
• 4. B. major
• 5. B. ovata
• 6. B. occultans
• 7. B. jakimovi .

•
• B. jakimovi, was discovered in Russia and does not
appear in many descriptions of Babesia.

• Sheep and goats:

• B. motasi
• B. ovis
 Morphology
• Babesia spp. are unicellular eukaryotes
• Taxonomically classified as Piroplasmida
• Commonly referred to as piroplasmids
• Prsent within the Apicomplexa group, together with
Theileria spp. and Cytauxzoon spp.
• Piroplasmids have been defined as tick transmitted,
piriform, round, or rod-shaped parasites that lack c
onoids and flagella in all life stages; without oocyst
s
• All ruminant-infecting Babesia species belong to t
he sensu stricto group
• This group is characterized by
• 1.Lack of a schizont stage
• 2. The occurrence of transovarial transmission in t
he tick
• These two phenotypic features that clearly disting
uish them from Theileria parasites
• Babesia spp. only invade erythrocytes within their
vertebrate hosts
• After entering an erythrocyte, the infectious sporoz
oite acquires a cyclic structure—a trophozoite ring
• Which then transforms into a replicative merozoite
• Merozoites are tear-shaped and normally observed
in pairs
• Although Maltese cross structures can occasionally
be found
Babesia divergens
EPIDEMIOLOGY
 Geographic Distribution
• The distribution of the babesia is governed by the
"geographic and seasonal distribution of the insec
t vectors that transmit them"
Distribution
 Species Affected
• Cattle are the primary hosts and reservoirs for B. b
ovis, B bigemina and B. divergens
• B. divergens have been identified in infected reinde
er (Rangifer tarandus)
• Experimentally infected, splenectomized non-huma
n primates (including chimpanzees, Pan troglodytes
and rhesus macaques, Macaca mulatta) become ill
• But spleen-intact primates were unaffected.
• Mongolian gerbils are the only laboratory rodents t
hat are readily infected with B. divergens without re
moving the spleen
• B. bovis and B bigemina have been detected in wat
er buffalo (Bubalus bubalis)
• Wild African buffalo (Syncerus caffer)
• African buffalo
• American bison (Bison bison)
• Wild white-tailed deer (Odocoileus virginianus) also
get affected
• B. major, B. ovata, and B. occultans are primarily
thought to infect cattle
• B. occultans have been reported in wild African b
uffalo
• B. major can cause clinical signs in experimentally
infected American bison
• B. jakimovi has been reported to infect cattle, ro
e deer, Asian elk (Alces alces), and reindeer
Interrelation between Babesia pathogen, verteb
rate host, and tick vector in bovine babesiosi
s epidemiology.
Life Cycle and Deve
lopment of
Babesia
• The development of B. bovis B. bigemina follows si
milar patterns
• Babesia spp. do not parasitize any vertebrate host c
ell other than erythrocytes
• Each sporozoite penetrates the erythrocyte membr
ane with the aid of a specialized structure called the
apical complex.
• Once inside, it transforms into a trophozoite
• From trophozoite two merozoites develop by a pro
cess of merogony or schizogony (binary fission)
• The tick becomes infected when ingesting erythroc
ytes containing piroplasms (gametocytes)
• They develop into male and female gametes in the
tick gut
• The microgametes fuse with macrogametes to pro
duce motile zygote
• The zygotes then multiply, and the resultant “verm
icules” invade numerous organs of the tick, includi
ng the ovaries.
• Therefore the infection passes through the ovary a
nd the egg to the next tick generation (called trans
ovarial transmission).
• Usually the female tick becomes infected
• Sporogony takes place in the salivary glands of larv
al, nymphal, and/or adult ticks of the next generati
on
• When the tick attaches to a new host, sporozoites
mature
• Example: In B. bigemina some development occurs
in the feeding larvae
• But infective sporozoites take about 9 days to appe
ar
• So therefore transmission occur in the nymphal and
adult stages of the tick.
• For B. bovis, the formation of infective sporozoites
usually occurs within 2 to 3 days of larval tick attac
hment
• The host is infected with infected tick saliva.
• Particular species of Babesia can persist for several
tick generations, even in the absence of new infecti
ons.
Other means of transmission:
• Contaminated needles
• Surgical instruments
• The ease with which infection can be
Life Cycle of Babesia bigemina
Origin of Infection
and Transmission
• Viable stages of Babesia are present in the bloodstr
eam of animals in the active phaseof the infection
• Ticks are the natural vectors of babesiosis
• The causative parasites persist and pass through pa
rt of their life cycle in the invertebrate host
• Both B. bovis and B. bigemina pass through part of
their life cycle in the tick Rhipicephalus australis

• Other species of Boophilus and Rhipicephalus are

major vectors of babesiosis
• Other ticks of the genera Haemaphysalis, Hyalom
ma, Ixodes, and Dermacentor can be involved in t
ransmission
• R. microplus and R. annulatus are one-host ticks t
hat complete their life cycle on a single host
• Ixodes ricinus is the major vector for B. divergens
• All three of its life stages are thought to be capabl
e of transmitting this organism
• Haemaphysalis longicornis transmits B. ovata
• B. occultans is thought to be transmitted by Hyalo
mma marginatum, Hy. rufipes
• B. major are thought to include Haemaphysalis pu
nctata
• B jakimovi might be transmitted by a member of th
e genus Ixodes.
• Cattle that have recovered from acute babesiosis
can remain asymptomatically infected, and recrud
escence of infection
• Parasitemia can occur at irregular intervals.
• Persistent infection with B. divergens, with period
ic waves of parasitemia, was also detected in infe
cted sheep
• Other means of transmission
• Transmitted during transfusions
• Smaller amounts of blood are transferred on reused
needles or field surgical instruments
• Biting flies
• Transplacental transmission has been demonstrate
d for B. bovis and B. bigemina in cattle
• But seems to be infrequent
Persistent Babesia Infectio
ns
• The outcome depends on the balance between ho
st and parasite responses tothe inflammatory resp
onse induced by the parasite
• After acute infection, animals either die or recover
from the infection
• Recovery is either naturally or after drug treatment
• After recovery, the parasite establishes persistency
and the host become chronically infected
• Following a tick bite, at the site of infection, an inn
ate immune response against infected erythrocyte
s or free Babesia merozoites is initiated
• Different Babesia pathogen associated molecular p
atterns (PAMPs) bind to mammalian toll-like recept
ors (TLRs)
• TLRs expressed on neutrophils or monocytes
• This is the process responsible for the initiation of th
e host innate immune response.
• Mainly, non-methylated CpG motifs of DNA bind to
bovine TLR9 Monocytes or phagocytic neutrophils
• Results in release of cytokines such as IL-B, TNF-α, a
nd Nitric Oxide (NO) in Babesia infections
• This chemokines attract immature dendritic cells (i
DC)
• Also, Natural Killer cells (NK) have been identified i
n the earlystages of Babesia infection
• NKs release interleukin (IL) IL-12, IL-18, and IFNγ
• Mature DC (mDC) migrate to secondary lymphoid
tissue, such as the spleen or lymph nodes, at the b
eginning of the acute infection
• Which is characterized by high parasitemia, fever,
anemia, drop of hematocrit, and organ failure
• In the spleen, the mDC present Babesia antigens to
naïve T cells
• Spleen macrophages activated by IFNγ kill the para
site by phagocytosis and the activity of toxic metab
olites such as NO
Immune regulation of Babesia i
nfection toward Persistence. D
Risk Factors
 Host Factors
• B. indicus breeds of cattle are considerably more r
esistant to babesiosis than B. taurusbreeds
• Zebu and Afrikaner cattle have a higher resistance
to B. bovis than British and European breeds
• Santa Gertrudis and crossbred cattle occupy an int
ermediate position
• Zebu-type cattle are also relatively free from the dis
ease because of their resistance to heavy tick infest
ation
• Age Resistance:
• In cattle, there is a variation in susceptibility to infe
ction according to age
• Severity of babesiosis increases with age.
• Calves from naïve dams are highly susceptible to i
nfection
• Clinical illness from birth to 2 months of age, at wh
ich time they develop resistance that persists to ap
proximately 6 months of age.
• Calves from immune dams receive antibodies via th
e colostrum
• This passive immunity persists for 3 to 4 months aft
er birth
• The highest infection rate is in animals between 6 to
12 months of age
• Infection is uncommon in animals of greater than 5
years of age
• Animals of less than 1 year of age are infected pre
dominantly with B. bigemina
• Animals greater than 2 years of age by B. bovis
 Environmental Factors
• There is seasonal variation in the prevalence of babe
siosis
• The greatest incidence occurring soon after the peak
of the tick population
• Of the climatic factors, environmental temperature is
the most important
• Because of its effect on tick activity—higher temperat
ures increase activity
• Humidity and rainfall have little effect
 Pathogen Factors

• Many intraerythrocytic hemoparasites survive host i

mmune responses through antigenic drift or shift
• This has been demonstrated for Babesia bovis
• There are different “strains” and antigenic variants
of both B. bovis and B. bigemina
• Babesia infections in cattle can relate to superinfecti
on with antigenically distinct parasite populations.
•
• Antigenic change can provide Babesia with a tempo
rary respite from attack by the host immune system
• So it might prolong the infection period or cause dis
ease relapses.
 Incubation Period

• Clinical signs usually appear 2-3 weeks after a bite

• from an infected tick. After inoculation with contam
inated
• blood, the incubation period can be as short as 4-5
days
• for B. bigemina and 10-12 days for B. bovis.
Pathogenesis
• Infection of a vertebrate host is initiated by inocul
ation of sporozoite stage into the bloodstream
• Most Babesia sporozoites directly invade circulati
ng erythrocytes.
• Once erythrocyte invasion occurs, a perpetual cycl
e of asexual reproduction is established
 Acute Cases
• When an animal becomes infected, multiplication o
f the protozoa in the visceral (B. bovis) or periphera
l (B. bigemina) vessels occurs
• It reaches a peak with the development of clinically
detectable hemolysis, the principal pathogenic eff
ect, after 7 to 20 days
• The hemolysis results in profound anemia, jaundic
e, and hemoglobinuria
• A fatal outcome as a result of anemic anoxia comm
only follows
• In surviving animals, there are ischemic changes in
skeletal and heart muscle
 B. bovis
• In B. bovis infection, there is also profound vasodila
tion and hypotension
• Its resulting from the stimulation of production of v
asoactive substances
• These substances associated with increase in vascul
ar permeability
• Circulatory stasis and shock follow; disseminated in
travascular coagulation (DIC)
• Subsequent fatal pulmonary thrombosis are also fea
tures
• Cerebral babesiosis is possible
• B. bigemina
• B. bigemina is an uncomplicated hemolytic agent
• It does not exert these vascular and coagulation eff
ects
• Susceptibility to infection with Babesia spp. decrea
ses with age
• But the severity of disease increases. For
• Example:
• Calves of 5 to 6 months of age infected with B. bov
is show limited clinical signs,
• Cattle of 1 to 2 years of age have a moderately sev
ere disease
• Aged cows suffer a severe, often fatal disease.
• Intrauterine infection with B. bovis has been repor
ted
• Animals that survive become carriers
• Carrier is a state in which a subclinical infection is ma
intained by a delicate immunologic balance betwee
n protozoa and antibodies
• This balance is readily disturbed by the stress of tran
sport, deprivation of food, pregnancy, or intercurrent
disease
•
• Carrier animals are resistant to infection with B. bo
vis for up to 2 years
• With constant reinfection, such as occurs in enzooti
c situations, protection is continuous
• The ability of cattle to infect ticks is much longer (1
year) with B. bovis than B. bigemina(4–7 weeks)
• Similarly, the peak incidence is at a younger age for
B. bigemina
• Reinfection rate is more rapid in young animals
• In pregnant cows, there is usually no apparent infe
ction of the calf in utero
• But passive immunity is transferred via colostrum t
o the newborn calf
CLINICAL FINDINGS
 Cattle
• Babesia Bovis:
• The acute disease generally runs a course of 3 to 7 d
ays
• Fever of greater than 40°C (104°F) is usually present
for several days before other signs emerge
• This is followed by inappetence, depression, polypne
a, weakness, and a reluctance to move.
• Hemoglobinuria is often present known as “redwat
er” in some countries
• Urine is dark red to brown in color and produces a
very stable froth
• Anemia and jaundice develop, particularly in prolo
nged and severe cases
• Diarrhea may occur.
• Muscle wasting, tremors, and recumbency develop
in advanced cases, followed terminally by coma
•.

• Cattle showed hemoglobinuria (A), poor body cond

ition (B), pale mucous membrane of eye(C) andpal
e mucous membrane of vaginadue tobabesiosis (D)
• Many severely affected animals die precipitately at
this point, after an illness of only 24 hours
• Metabolic acidosis can be present in a significant p
ercentage of cases of bovine babesiosis
• During the fever stage, pregnant cattle can abort
• Bulls may become sterile for 6 to 8 weeks.
• Cerebral babesiosis is manifested by
• Incoordination
• Followed by posterior paralysis
• Mania
• Convulsions
• Coma
• The mortality rate in such cases is high, in spite of c
hemotherapy
• In those that survive, the febrile stage usually lasts
for approximately a week
• The total course about 3 weeks
• Cattle that survive recover gradually from the sever
e emaciation and anemia, which are inevitable se
quelae.
 Subacute Syndrome
• Particularly in young calves
• Fever is mild and hemoglobinuria is absent.
• The syndrome associated with B. divergens is simila
r to that of B. bovis
• Exception is that, in addition, there is spasm of the
anal sphincter
• Causing the passage of feces with great force in a lo
ng, thin stream, even in the absence of diarrhea; thi
s sign is referred to as “pipe-stem” feces.
 Babesia Bigemina
• Hemoglobinuria is present earlier and more consis
tently than inB. bovis infection
• Fever is less of a feature
• Acutely affected animals are usually not as severely
affected as those with B. bovis infection
• There is no cerebral involvement
• Recovery in nonfatal cases is usually rapid and comp
lete
• However, in some cases, disease can develop very r
apidly, with sudden and severe anemia, jaundice, a
nd death.
• Animals that recover from B. bigemina remain infec
tive to ticks for 4 to 7 weeks and remain as carriers f
or only a few months
 Differential diagnosis
• Anaplasmosis
• Trypanosomiasis
• Theileriosis
• Bacillary hemoglobinuria
• Leptospirosis
• Eperythrozoonosis
• Rapeseed poisoning
• Chronic copper poisoning
• Rabies and other encephalitides may also be consid
erations in cattle with central nervous system signs.
Diagnosis
 Clinical diagnosis
• Clinical manifestations of disease associated with B
B are typical of a haemolytic anaemia disease proc
ess
• But vary according to agent (i.e. species of parasit
e) and host factors (i.e. age, immune status)
• BB is predominantly observed in adult cattle with
B. bovis being more pathogenic than B. bigemina
or B. divergens
• Infected animals develop a life-long immunity again
st re-infection with the same species and some cro
ss-protection
Babesia bovis
 High fever
 Ataxia and incoordination
 Anorexia
 Production of dark red or brown-colored urine
 Signs of general circulatory shock
 Sometimes nervous signs associated with sequestra
tion of infected erythrocytes in cerebral capillaries
  Anaemia and haemoglobinuria may appear later in
the course of the disease
 In acute cases: maximum parasitaemia (percentage
of infected erythrocytes) in circulating blood is often l
ess than 1%
Babesia bigemina
 Fever
 Haemoglobinuria and anaemia
 Production of dark red or brown-colored urine
 Nervous signs minimal or non-existent as intravascul
ar sequestration of infected erythrocytes does not occ
ur
 Parasitaemia often exceeds 10% and may be as high
as 30%
 Lesions
• Lesions observed are those most often associated
with an intravascular haemolytic condition
• Pale or icteric mucous membranes; blood may app
ear thin and watery
• Subcutaneous tissues, abdominal fat and omentum
may appear icteric
• Swollen liver with an orange-brown or paler colora
tion; enlarged gall bladder containing thick, granula
r bile
• Enlarged, dark, friable spleen
• Kidneys appear darker than normal with possible pe
techial haemorrhages
• Bladder may contain dark red or brown-colored uri
ne
• Possible oedema of lungs
• Petechiae or ecchymoses on surface of heart and br
ain
Opened urinary bladder of a bovine, show
ing haemoglobinuria
Bovine, brain. The cerebral cortex is dif
fusely reddened (“cerebral flush”).
 Samples
1. Several thick and thin blood smears collected from
superficial skin capillaries
• Example: Tip of the ear or tip of the tail of live anim
als during the acute phase of the disease (appearan
ce of fever)
2. Sterile jugular blood
• Should be collected into an anticoagulant such as l
ithium heparin or ethylene diamine tetra-acetic aci
d (EDTA)
3.From dead animals:
• Thin blood films, as well as smears from organs
• Organ smears acquired at necropsy: cerebral cortex
, kidney (freshly dead), spleen (when decompositio
n is evident), heart muscle, lung and liver
4.Babesia parasites can sometimes be detected in ca
pillary blood taken from the lower limb
region one or more days after death
5. Serum samples should also be collected
Identification of the agent
1.Microscopic examination of blood –
• Traditional method of identifying agent in infected
animals by microscopic examination of Giemsa-stai
ned thick and thin blood films
• Sensitivity of thick films can detect parasitaemias a
s low as 1 parasite in 106 red blood cells
• Babesia species differentiation is good in thin films
but poor in the more sensitive thick films
• Adequate for detection of acute infections
• But not for detection of carriers where parasitaemi
as are very low
• Parasite identification and differentiation improved
by using a fluorescent dye, such as acridine orange
instead of Giemsa
2.Nucleic acid-based diagnostic assays -
• Very sensitive particularly in detecting B. bovis and
B. bigemina in carrier cattle
• PCR- based techniques are reported to be at least 1
000 times more sensitive than thin blood smears fo
r detection of B. bovis
• A number of PCR techniques have been described t
hat can detect and differentiate species of Babesia i
n carrier infections
3.In-vitro culture methods:
• Used to demonstrate presence of carrier infections
of Babesia spp.; B. bovis has also been cloned in cu
lture
• Minimum parasitaemia detectable by this method
• Could be as low as 10–10,making it a very sensitive
method for the demonstration of infection, with 10
0% specificity
 Serological tests
1. Babesia bovis enzyme-linked immunosorbent assa
y
o ELISA for diagnosis of B. bovis infection uses a whole me
rozoite antigen
o Competitive ELISAs using recombinant merozoite surfac
e and rhoptry associated antigens of B. bovis have recent
ly been developed
o Reduction in specificity of the indirect B. bovis ELISA usi
ng recombinant antigens has been noted in some situatio
ns
2.Babesia bigemina enzyme-linked immunosorbe
nt assay
• A competitive ELISA developed and validated
• No other well-validated ELISA available for B. bige
mina
• ELISAs have also been developed for B. divergens u
sing antigen derived from culture, Meriones or catt
le
• But none has been validated internationally
3.Indirect fluorescent antibody (IFA) test
• Widely used in the past to detect antibodies to Babe
sia spp.
• But the B. bigemina test has poor specificity
• Cross-reactions with antibodies to B. bovis in the B. b
igemina
• IFA test are a particular problem in areas where the t
wo parasites coexist
4.Complement fixation
• Has been used to detect antibodies against B. bovis
and B. bigemina
• Used to qualify animals for importation into some c
ountries
PREVENTION AND CONT
ROL
 Sanitary prophylaxis
1. Eradication of BB (Bovine Babesiosis) has been a
ccomplished by elimination of tick vector and/or i
ntensive chemotherapeutic regimes
• In areas where eradication of tick is not feasible o
r desirable, ticks are controlled by repellents and
acaricides
2.Reducing exposure of cattle to ticks
o Repellents, acaricides and regular inspection; animal
s and premises
o Control and eradication of the tick vector

3.Cattle develop a durable, long-lasting immunity aft

er a single infection with B. bovis, B. divergens or B. bi
gemin
4.Endemic environments should be monitored car
efully
• Introduction of immuno-naïve animals
• Introduction of new species or strains of disease age
nt
• Interruptions in exposure to ticks and disease due to
changes in climate, host factors and management
 Medical prophylaxis
1. Vaccine for Babesia:
• Live vaccine: Most live vaccines contain specially sel
ected strains of Babesia (mainly B. bovis and B. big
emina)
• These are produced in calves or in vitro in governm
ent-supported production facilities as a service to th
e livestock industries
• Caution should be used in their employment as the
y may be virulent in adult animals
• May be contaminated with other disease agents an
d could lead to hypersensitivity reactions
• Usually used in younger animals
• An experimental B. divergens vaccine prepared fro
m the blood of infected Meriones has also been us
ed successfully
2. Killed vaccine: prepared from blood of B. diverg
ens-infected calves
3. Other vaccines:
• Despite the worldwide efforts, the prospects for rec
ombinant vaccines against Babesia spp. Remain ch
allenging
• To date, no effective subunit vaccine is available co
mmercially
• Endemic areas
• Clinically affected animals treated with an antiparasi
tic drug (diminazene diaceturate, imidocarb, amicarb
alide)
• Efficacy depends on timely detection early in disease
• Babesia parasites can be cleared from carrier animal
s
• Imidocarb has been reported to protect animals fro
m disease but allow development of immunity
• Caution in regard to residues in milk and meat
Treatment
• firstly, treatment with a babesicide
• Secondly, the need for supportive therapy such as b
lood transfusion and fluid replacement.
• There are few babesicides now available.
• Imidocarb, which is given at a dose rate of 1mg/kg b
ody weight
• It is highly effective and relatively non-toxic, but do
es have tissue residues for several weeks after its us
e.
• It can also be used at twice the therapeutic dose as
a chemoprophylactic
• Giving protection from infection for up to six weeks
• Its used in this way to administer to cattle
• 1. That will be exposed to ticks
• 2. That have been vaccinated with live Babesia with t
he hope that cattle will become mildly affected but p
rotected from clinical illness
• 3. Immunity to further infection will be stimulated
 Supportive Theraphy
• Blood transfusion is frequently required for severel
y affected adult cattle
• So normally achieved by collection of 5 l of blood fr
om an unaffected healthy cow into a 22 per cent so
lution of the anticoagulant acid citrate dextrose (AC
D)
• The mixture being immediately transfused into the
recipient animal.
• Such single transfusions without cross-matching of
blood are usually successful
• But repetition can lead to problems of incompatibili
ty of blood antigens.
Thank you...........