College of Veterinary and Animal Sciences, Parbhani

DEPARTMENT OF VETERINARY PATHOLOGY

TOPIC: ANAPLASMOSIS, EHRLICHIOSIS & Q FEVER

SUBMITTEDTO: DR.B.M. KONDRE

Assistant Professor
Department of veterinary pathology

SUBMITTED BYKAUSTUBH V.ANTURKAR

REG.NO.-13V02B
BATCH:A

ANAPLASMOSIS
Anaplasmosis, formerly known as gall sickness, traditionally refers to a disease of
ruminants caused by obligate intraerythrocytic bacteria of the order Rickettsiales,
family Anaplasmataceae, genus Anaplasma. Cattle, sheep, goats, buffalo, and some
wild ruminants can be infected with the erythrocytic Anaplasma. Anaplasmosis occurs
in tropical and subtropical regions worldwide (~40N to 32S), including South and
Central America, the USA, southern Europe, Africa, Asia, and Australia.The Anaplasma
genus also includes A phagocytophilum ,A bovis, andA platys,all of which invade blood
cells other than erythrocytes of their respective mammalian hosts. Bovine
anaplasmosis is of economic significance in the cattle industry.

Etiology and Pathogenesis

Clinical bovine anaplasmosis is usually caused by A marginale. An A marginale with an
appendage has been called A caudatum, but it is not considered to be a separate
species. Cattle are also infected with A centrale, which generally results in mild disease.
A ovis may cause mild to severe disease in sheep, deer, and goats. A phagocytophilum
has recently been reported to infect cattle; however, natural infection is rare and it
does not cause clinical disease.

Transmission and Epidemiology

Up
to
17
different
tick
vector
species
(including
Dermacentor,Rhipicephalus,Ixodes,Hyalomma, and Argas) have been reported to
transmit Anaplasmaspp. Notall of these are likely significant vectors in the field, and it
has been shown that strains of A marginale also coevolve with particular tick strains.
Rhipicephalus (Boophilus) spp are major vectors in Australia and Africa, and
Dermacentor spp have been incriminated as the main vectors in the USA. After feeding
on an infected animal, intrastadial or trans-stadial transmission may occur.
Transovarial transmission may also occur, although this is rare, even in the single-host
Rhipicephalus spp. A replicative cycle occurs in the infected tick. Mechanical
transmission via biting dipterans occurs in some regions. Transplacental transmission
has been reported and is usually associated with acute infection of the dam in the
second or third trimester of gestation. Anaplasmosis may also be spread through the
use of contaminated needles or dehorning or other surgical instruments. There is a
strong correlation between age of cattle and severity of disease. Calves are much more
resistant to disease (although not infection) than older cattle. This resistance is not due
to colostral antibody from immune dams. In endemic areas where cattle first become
infected with A marginale early in life, losses due to anaplasmosis are minimal. After
recovery from the acute phase of infection, cattle remain chronically infected carriers
but are generally immune to further clinical disease. However, these chronically

infected cattle may relapse to anaplasmosis when immune suppressed (eg, by

corticosteroids), when infected with other pathogens, or after splenectomy.

Clinical Findings

In animals <1 yr old anaplasmosis is usually subclinical, in yearlings and 2-yr-olds it is

moderately severe, and in older cattle it is severe and often fatal. Anaplasmosis is
characterized by progressive anemia due to extravascular destruction of infected and
uninfected erythrocytes. The prepatent period ofA marginaleis directly related to the
infective dose and typically ranges from 1536 days (although it may be as long as 100
days). After the prepatent period, peracute (most severe but rare), acute, or chronic
anaplasmosis may follow. Rickettsemia approximately doubles every 24 hr during the
exponential growth phase. Generally, 10%30% of erythrocytes are infected at peak
rickettsemia, although this figure may be as high as 65%. RBC count, PCV, and
hemoglobin values are all severely reduced. Macrocytic anemia with circulating
reticulocytes may be present late in the disease.Animals with peracute infections
succumb within a few hours of the onset of clinical signs. Acutely infected animals lose
condition rapidly. Milk production falls. Inappetence, loss of coordination,
breathlessness when exerted, and a rapid bounding pulse are usually evident in the
late stages. The urine may be brown but, in contrast to babesiosis, hemoglobinuria
does not occur. A transient febrile response, with the body temperature rarely
exceeding 106F (41C) occurs at about the time of peak rickettsemia. Mucous
membranes appear pale and then yellow. Pregnant cows may abort. Surviving cattle
convalesce over several weeks, during which hematologic parameters gradually return
to normal.Bos indicusbreeds of cattle appear to possess a greater resistance toA
marginaleinfection thanB taurusbreeds, but variation of resistance of individuals within
breeds of both species occurs. Differences in virulence betweenAnaplasmastrains and
the level and duration of the rickettsemia also play a role in severity of clinical
manifestations.

Lesions

Lesions are typical of those found in jaundiced. Blood is thin and watery. The spleen is
characteristically enlarged and soft, with prominent follicles. The liver may be mottled
and yellow-orange. The gallbladder is often distended and contains thick brown or
green bile. Hepatic and mediastinal lymph nodes appear brown. There are serous
effusions in body cavities, pulmonary edema, petechial hemorrhages in the epi- and
endocardium, and often evidence of severe GI stasis. Widespread phagocytosis of
erythrocytes is evident on microscopic examination of the reticuloendothelial organs.A
significant proportion of erythrocytes are usually found tobe parasitized after death
due to acute infection

Diagnosis
Anaplasma marginale, bovine blood smearA marginale, together with the
hemoprotozoa Babesia bovis and B bigemina, are the causative agents of tick fever in
cattle. These three species have similar geographic distributions, except that an
aplasmosis occurs in the absence of babesiosis in the USA. Microscopic examination of
Giemsa-stained thin and thick blood films is critical to distinguish anaplasmosis from
babesiosis and other conditions that result in anemia and jaundice, such as
leptospirosis and theileriosis.Blood in anticoagulant should also be obtained for
hematologic testing. In Giemsa-stained thin blood films, Anaplasma spp appear as
dense, homogeneously staining blue-purple inclusions 0.31.0 m in diameter. A
marginale inclusions are usually located toward the margin of the infected erythrocyte,
whereas A central inclusion bodies are located more centrally. A caudatum cannot be
distinguished from A marginale using Giemsa-stained blood films. Special staining
techniques are used to identify this species based on observation of characteristic
appendages associated with the bacteria.A caudatum has been reported only in North
America and could possibly be a morphologic form of A marginale and not a separate
species. Inclusion bodies contain 18 initial bodies 0.30.4 m in diameter, which are
the individual rickettsiae. The percentage of infected erythrocytes varies with the stage
and severity of disease; maximum rickettsemias in excess of 50% can occur with A
marginale. Microscopically, the infection becomes visible 26 wk after transmission.
During the course of infection, the rickettsemia can double each day for up to 10 days
and then decreases. Severe anemia can persist for weeks after parasites cannot be
detected in blood smears. Chronically infected carriers may be identified with a fair
degree of accuracy by serologic testing using the msp5 ELISA, complement fixation, or
card agglutination tests.

Treatment, Control, and Prevention

Tetracycline antibiotics and imidocarb are currently used for treatment. Cattle may be
sterilized by treatment with these drugs and remain immune to severe anaplasmosis
subsequently for at least 8 mo.Prompt administration of tetracycline drugs
(tetracycline,
chlortetracycline,
oxytetracycline,
rolitetracycline,
doxycycline,
minocycline) in the early stages of acute disease (eg, PCV >15%) usually ensures
survival. A commonly used treatment consists of a single IM injection of long-acting
oxytetracycline at a dosage of 20 mg/kg. Blood transfusion to partially restore the PCV
greatly improves the survival rate of more severely affected cattle. The carrier state
may be eliminated by administration of a long-acting oxytetracycline preparation (20
mg/kg, IM, at least two injections with a 1-wk interval). Withholding periods for
tetracyclines apply in most countries. Injection into the neck muscle rather than the
rump is preferred.Imidocarb is also highly efficacious against A marginale as a single
injection (as the dihydrochloride salt at 1.5 mg/kg, SC, or as imidocarb dipropionate at
3 mg/kg). Elimination of the carrier state requires the use of higher repeated doses of

imidocarb (eg, 5 mg/kg, IM or SC, two injections of the dihydrochloride salt 2 wk apart).
Imidocarb is a suspected carcinogen with long withholding periods and is not approved
for use in the USA or Europe. In South Africa, Australia, Israel, and South America,
infection with live A centrale(originating from South Africa) is used as a vaccine to
provide cattle with partial protection against the disease caused by A marginale. A
centrale(single dose) vaccine produces severe reactions ina small proportion of cattle.

Q FEVER

Coxiellosis is a zoonotic bacterial infection associated primarily with parturient

ruminants, although domestic animals such as cats and a variety of wild animals have
been identified as sources of human infection. The human zoonotic infectious process
associated with Coxiella burnetiiis widely known as Q fever. Coxiella is considered a
potential agent of bioterrorism because of its low infectious dose, stability in the
environment, and capability for aerosol dispersion.

Etiology, Epidemiology, and Transmission

Coxiellosis is caused by the gram-negative coccobacillus C burnetii. Although classically
considered a rickettsial agent, recent phylogenetic analyses suggest that C burnetiiis
more closely related to Legionella and Francisella than to the genus Rickettsia. It
resides and reproduces in the acidified phagolysosomes of host monocytes and
macrophages. Two forms exist: the large cell variant is a vegetative form found in
infected cells, and the small cell variant is the extracellular infectious form shed in milk,
urine, and feces and found in high concentration (109ID50/g) in placental tissue and
amniotic fluid. The small cell variant is resistant to heat, drying, and many common
disinfectants and remains viable for weeks to years in the environment. Once a
domestic ruminant is infected, C burnetiican localize in mammary glands,
supramammary lymph nodes, placenta, and uterus, from which it may be shed in
subsequent parturitions and lactations. The epidemiology of C burnetiiis complex
because there are two major patterns of transmission. In one, the organism circulates
between wild animals and their ectoparasites, mainly ticks; the other occurs in
domestic ruminants, independent of the wild animal cycle. Ixodid andargasid ticks can
act as reservoirs of the organism. Distribution is worldwide (except New Zealand), and
the host range includes various wild and domestic mammals, arthropods, and birds.
The disease is enzootic in most areas where cattle, sheep, and goats are kept. In the
USA, seroprevalence studies have shown significant variability in prevalence of
antibodies to C burnetii based on difference in test population, test used, and time of
year. Evaluation of bulk tank milk sampling of USA cattle dairies has demonstrated
prevalence of the organism at the farm level of 77% to >90%. In a comprehensive
seroprevalence study conducted in Canada, 48.6% of sheep operations and63.2% of
goat operations had at least one positive animal. Not surprisingly, sero prevalence in

veterinarians and small ruminant farmers is also high .The greatest risk of transmission
occurs at parturition by inhalation, ingestion, or direct contact with birth fluids or
placenta. The organism is also shed in milk, urine, and feces. High-temperature
pasteurization effectively kills the organism. Ticks may transmit the disease among
domestic ruminants but are not thought to play an epidemiologically important role in
transmission of disease to people.

Clinical Findings and Diagnosis

Infection in ruminants is usually subclinical but can cause anorexia and late abortion.
When infection is subclinical, animals shed much lower bacterial loads of the organism
than when abortion occurs. Reports have implicated C burnetii as a cause of infertility
and sporadic abortion with a necrotizing placentitis in ruminants. New evidence has
shown an association of C burnetii with subclinical mastitis among dairy cows,
although additional work regarding causality is required before this can be considered
clinically valid. Experimental infection in cats causes transient fever, dullness, and
anorexia lasting several days. In domestic ruminants, gross lesions are nonspecific, and
differential diagnosis should include infectious and noninfectious agents that cause
abortion. Immunofluorescence test on paired sera taken 2 wk apartcan be used to
detect recent infection; however, shedding of C burnetii may occur in the absence of a
measurable serum antibody titer in up to 20% of infected animals. Culture,
immunohistochemical, and PCR tests may be used to identify C burnetii in tissues.
Studies conducted in veterinary diagnostic laboratories suggest that C burnetii is often
found concurrently with other organisms isolated in cases of infectious abortions, so
mixed infections may be important. Seasonal variability in shedding of the organism
hinders interpretation of a single PCR test. Shedding is highest in the periparturient
period and may drop below detectable levels for a significant period during the year
despite persistent infection.

Treatment and Control

Q fever in people is a notifiable disease in the USA, primarily because of its status as a
possible bioterrorism agent; reporting requirements for animals vary by state. Vaccines
for people and animals have been developed but are not commercially available in the
USA. Vaccination has prevented infection when administered to uninfected calves and
has improved fertility and reduced shedding in previously infected animals. There is
little evidence-based data to suggest that antibiotic treatment in animals provides
significant benefit. Human clinical disease is typically treated with tetracyclines, but
significant benefit from tetracycline treatment has not been demonstrated in
controlled studiesin abortion outbreaks of sheep in Europe. Despite the lack of
evidence, some practitioners still advocate the use of parenteral tetracyclines during
abortion storms. In known infected herds, the periparturient period represents a

significant risk period for transmission because of the large amount of environmental
contamination associated with abortion. Standard abortion control measures,
including prompt removal of aborted materials (using zoonotic precautions),
segregation of animals by pregnancy status, and diagnostic evaluation of abortions, are
all warranted. Zoonotic Risk Q fever occurs more frequently in persons who have
occupational contact with high-risk species. The clinical presentation in people is highly
variable clinically, ranging from a self-limiting, influenza-like illness to pneumonia,
hepatitis, and endocarditis. C burnetii is highly infectious, and a single organism can
reportedly cause infection via the aerosol route in people. ed milk. Handling of infected
tissue poses a threat to laboratory personnel.

Signs and symptoms

Incubation period is usually two to three weeks.The most common manifestation is flulike symptoms with abrupt onset of fever,malaise,profuse perspiration, severe
headache,muscle pain,joint pain,loss of appetite, upper respiratory problems, dry
cough,pleuritic pain, chills,confusion and gastrointestinal symptoms, such as nausea,
vomiting, and diarrhea .Approximately half of infected individuals exhibit no
symptoms. During its course, the disease can progress to anatypical pneumonia, which
can result in a life-threatening acute respiratory distress syndrome(ARDS), whereby
such symptoms usually occur during the first four to five days of infection.[citation
needed]Less often, Q fever causes (granulomatous)hepatitis, which may be
asymptomatic or becomes symptomatic with malaise, fever,liver enlargement, and
pain in the right upper quadrant of the abdomen. Whereas transaminase values are
often elevated ,jaundiceis uncommon. Retinal vasculitis is a rare manifestation of Q
fever.The chronic form of Q fever is virtually identical to inflammationof the inner
lining of the heart (endocarditis),which can occur months or decades following the
infection. It is usually fatal if untreated. However, with appropriate treatment, the
mortality falls to around 10%.Clinical signs in animals Cattle, goats and sheep are most
commonly infected, and can serve as a reservoir for the bacteria. Q fever is a well
recognized cause of abortions in ruminants and in pets .C. burnetii infection in dairy
cattle has been well documented and its association with reproductive problems in
these animals has been reported in Canada, USA, Cyprus, France, Hungary, Japan,
Switzerland and West Germany.For instance, in a study published in 2008,a significant
association has been shown between the sero positivity of herds and the appearance
of typical clinical signs of Q fever such as abortion, stillbirth, weak calves and repeat
breeding. Moreover, experimental inoculation ofC. Burneti in cattle induced not only
respiratory disorders and cardiac failures (myocarditis) but also frequent abortions and
irregular repeat breedings.

Diagnosis is usually based onserology(looking for an antibody response) rather than

looking for the organism itself. Serology allows the detection of chronic infection by the
appearance of high levels of the antibody against the virulent form of the bacterium.
Molecular detection of bacterial DNA is increasingly used. Culture is technically difficult
and not routinely available in most microbiology laboratories. Q fever can cause
endocarditis(infection of the heart valves) which may require trans esophageal
echocardiography to diagnose. Q fever hepatitis manifests as an elevation of ALT and

AST, but a definitive diagnosis is only possible on liver biopsy, which shows the
characteristic fibrin ring granulomas.

Treatment

Treatment of acute Q fever with antibiotics is very effective[citation needed]and should

be given in consultation with an infectious diseases specialist.[citation
needed]Commonly
used
antibiotics
includedoxycycline,tetracycline,chloramphenicol,ciprofloxacin,ofloxacin, and hydroxyl
chloroquine. Chronic Q fever is more difficult to treat and can require up to four years
of treatment with doxycycline
and quinolones or
doxycycline
with
hydroxychloroquine.Q fever in pregnancy is especially difficult to treat because
doxycycline and ciprofloxacin are contraindicated in pregnancy. The preferred
treatment is five weeks of co-trimoxazole.

Prevention

Protection is offered by Q-Vax, a whole-cell, inactivated vaccine developed by an

Australian vaccine manufacturing company, CSL. The intradermal vaccination is
composed of killed Coxiella burnetii organisms. Skin and blood tests should be done
before vaccination to identify pre-existing immunity, because vaccinating subjects who
already have an immunity can result in a severe local reaction. After a single dose of

vaccine, protective immunity lasts for many years. Revaccination is not generally
required. Annual screening is typically recommended.

Ehrlichiosis

Etiology
Canine monocytic ehrlichiosis is usually caused by the rickettsia Ehrlichia canis,
although other types of Ehrlichia are sometimes involved. (Rickettsiae are a specialized
type of bacteria that live only inside other cells.) Carried by ticks, the organism infects a
certain type of white blood cell and causes fever and other signs. A related organism,
Ehrlichia ewingi, targets other types of white blood cells called granulocytes and has
been isolated from dogs and people in the southern, western, and midwestern United
States. Anaplasma platys, another rickettsia, causes infectious cyclic thrombocytopenia
in dogs. This infection leads to periodic losses of platelets, which causes problems with
blood clotting.The Ehrlichia and Anaplasma rickettsiae are present in many parts of the
world, including the United States. They are transmitted by ticks (including the brown
dog tick, lone star tick, and black-legged tick) that become infected after feeding on
infected animals. People, dogs,cats, and other domestic animals are accidental hosts of
these disease-causing organisms.

In infections caused by Ehrlichia canis, signs commonly progress from short to long
term, depending on the strain of the organism and the immune status of the host. In
short-term cases, there is fever, widespread inflammation of the lymph nodes,
enlargement of the spleen, and a decrease in the number of platelets in the
bloodstream. In addition, there may be loss of appetite, depression, loss of stamina,
stiffness and reluctance to walk, swelling of the limbs or scrotum, and coughing or
difficulty in breathing. Most short-term cases are seen in the warmer months, when
ticks are active. During this phase of infection, death is rare and the infected animal
may recover spontaneously. The recovered dog may remain free of signs thereafter, or
long term disease may develop.Long term ehrlichiosis caused by Ehrlichia canis may
develop in any breed of dog, but certain breeds (such as German Shepherds) may be
predisposed. Long term infection does not vary with the seasons. Signs depend on
which organs are affected and may include enlargement of the spleen, kidney failure,
and inflammation of the lungs, eye, brain and spinal cord. If the brain and spinal cord
are involved, there may be problems with the nervous system, such as lack of
coordination, depression, partial paralysis, and increased sensitivity to a normally
painless touch. Severe weight loss is common.Dogs infected with Anaplasma platys
generally show minimal to no signs of infection, although the organism is present in
the platelets. Infection with other rickettsiae causes signs similar to short-term
Ehrlichia canis infection, but the disease is usually more self-limiting. Fever and a
lameness that shifts from one leg to another may be present. Longterm disease (such
as that seen with Ehrlichia canis infection) is not typically seen in other ehrlichial

infections.For treatment of all forms of infection caused by these organisms, your

veterinarian will prescribe an antibiotic. The medication is usually given for a period of
10 to 21 days. Fever usually ends within 1 to 2 days after treatment begins. In long
term cases, the blood abnormalities may persist for 3 to 6 months, although relief from
signs often occurs much sooner. Some dogs may also need supportive care. If the dog
has widespread or severe bleeding, then transfusion with blood platelets or whole
blood may be prescribed. The most important preventive steps are those that control
ticks, the most common source of the disease. Keeping your dog away from areas
known to harbor ticks is a step you can take.

Diagnosis

Because thrombocytopenia is a relatively consistent finding with these infections, a

platelet count is an important screening test. Clinical diagnosis may be confirmed by
demonstrating the organisms within WBCs or platelets, seen in intracytoplasmic
inclusion bodies called morulae. This method of diagnosis lacks sensitivity, because low
numbers of organisms make demonstration difficult. More commonly, a diagnosis is
made by a combination of clinical signs, positive serum indirect fluorescent antibody
(IFA) titer, and response to treatment. In-house tests for E canis, A phagocytophilum, A
platys, and E ewingi Based on enzyme immunoassay methods are also available. The
antibody response may be delayed for several weeks; thus, serologic testing may not
be a reliablediagnostic tool early in the course of the disease. Furthermore, antibodies
can persist for months or years after infection, making in-house tests for the organisms
problematic for confirmation of acute infection, particularly in highly enzootic areas
where many dogs may have antibodies to these agents because of previous infections.
Testing of paired sera and demonstration of increased antibody titers is recommended
to confirm infection when possible, although treatment of suspected cases should
never be delayed or withheld on the basis of test results, either positive or negative.
Serologic cross-reactivity is strong between E canis, E chaffeens is, and E ewingii; some
cross-reactivity toA phagocytophilum is also seen. In people, the EML agent shows
cross-reactivity to E chaffeens is. In some areas, ~50% of dogs infected with E canis also
have a titer to A platys, which likely reflects coinfection; cross-reactivity between these
agents is not seen.PCR has been used to detect and identify specific Ehrlichia and
Anaplasma species in infected people and animals. Samples appropriate for PCR
include blood, tissue aspirates, or biopsy specimens of reticuloendothelial organs, such
as lymph nodes, spleen, liver, or bone marrow.PCR can also be used to detect the
effectiveness of treatment in clearing infection. PCR is not routinely available through
commercial veterinary laboratories, although some veterinary schools and research
institutions offer it. PCR is available through several commercial human laboratories.
During the acute stage, differential diagnoses include other causes of fever and
lymphadenomegaly (eg, Rocky Mountain spotted fever, brucellosis, blastomycosis,
endocarditis), immune-mediated diseases (eg, systemic lupus erythematosus), and
lymphosarcoma. During the chronic stage of E canis infection, differential diagnoses
include estrogen toxicity, myelophthisis, immune-mediated pancytopenia, and other
multisystemic diseases associated with specific organ dysfunction (eg,
glomerulonephritis).

Signs and symptoms

The most common symptoms include headache,muscle aches, and fatigue. A rash may
occur, but is uncommon. Ehrlichiosis can also blunt the immune system by suppressing
production of TNF-alpha, which may lead to opportunistic infections such as
candidiasis. Most of the signs and symptoms of ehrlichiosis can likely be ascribed to
the immune dysregulation that it causes. Late in infection, however, production of this
substance can be up regulated by 30 fold, which is likely responsible for the "toxic
shock-like" syndrome seen in some severe cases of ehrlichiosis. Some cases can
present with purpura and in one such case the organisms were present in such
overwhelming numbers that in 1991 Dr. Aileen Marty of the AFIP was able to
demonstrate the bacteria in human tissues using standard stains, and later proved that
the organisms were indeed Ehrlichia using immune peroxidase stains.Experiments in
mouse models further supports this hypothesis, as mice lacking TNF-alpha I/II
receptors are resistant to liver injury caused by ehrlichia infection.3% of human
monocytic ehrlichiosis cases result in death; however, these deaths occur "most
commonly in immune suppressed individuals who develop respiratory distress
syndrome, hepatitis, or opportunistic nosocomial infections.

Treatment

Doxycycline and minocycline are the medications of choice. For people allergic to
antibiotics of the tetracycline class, rifampin is an alternative. Early clinical experience
suggested that chloramphenicol may also be effective, however, in vitro susceptibility
testing revealed resistance.[citation needed.

Prevention

No human vaccine is available for ehrlichiosis. Tick control is the main preventive
measure against the disease. However, in late 2012 a breakthrough in the prevention
of CME (canine monocytic ehrlichiosis) was announced when a vaccine was accidentally
discovered by Prof. Shimon Harrus, Dean of the Hebrew University of Jerusalem's Koret
School of Veterinary Medicine.[