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Journal of Immunology and Immunopathology
Vol. 19, No. 2, July-December, 2017: 61-69
DOI: 10.5958/0973-9149.2017.00009.0
61
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A produ ct of Diva Ente rpr ises Pvt. Ltd .
Review Article
Brucellosis: A Current Review Update on Zoonosis
Shabu Shoukat1, Henna Wani2, Umer Ali1, Parveez Ahmad Para3, Shabnam Ara4 and
Subha Ganguly5*
1Ph.D. Research Scholar, Department of Veterinary Public Health, 2Ph.D. Research Scholar, Department of Veterinary
Pathology, Sher-e-Kashmir University of Agricultural Sciences and Technology-Kashmir (SKUAST-K), Kashmir-
190006, Jammu & Kashmir, India
3Assistant Professor, Department of Livestock Products Technology, 5Associate Professor, Department of Veterinary
Microbiology, Arawali Veterinary College (Affiliated with Rajasthan University of Veterinary and Animal Sciences,
Bikaner), N.H.-52 Jaipur Road, V.P.O. Bajor, Sikar-332001, Rajasthan, India
4PG Scholar, Department of Clinical Psychology, Indira Gandhi National Open University (IGNOU), New Delhi, India
*Corresponding author email id: ganguly38@gmail.com
Submission: 25-6-2017; Revision: 27-7-2017; Acceptance: 5-8-2017
ABSTRACT
Shoukat S, Wani H, Ali U, Para PA, Ara S and Ganguly S (2007). Brucellosis: A current review update on
zoonosis. J. Immunol. Immunopathol. 19(2): 61-69.
Brucellosis is an infectious, contagious zoonotic disease which is spread worldwide, caused by bacteria
of the genus Brucella. Brucellae are facultative intracellular parasites of the reticuloendothelial system and is
characterised by wave-like variations in the body temperature of afflicted victims. It was formerly known as
Mediterranean fever, Malta fever or undulant fever. Its modern name bears tribute to Sir David Bruce, the
military physician who discovered the aetiologic agent, Brucella melitensis. The disease primarily affects
cattle, sheep, goats, swine and dogs. The different Brucella spp. includes B. melitensis, Brucella abortus,
Brucella canis and Brucella suis, and their preferential hosts are sheep/goats, cattle/bison, dogs and pigs,
respectively. However, cross-species infections can occur; for example cattle can be affected by both B.
abortus and B. melitensis at the same time [Abdussalam and Fein (1976). Developments in Biological
Standardization, 31: 23]. Hence, from epidemiological evidence, the three species (B. abortus, B. melitensis
and B. suis) have distinct host preferences, and the organisms are capable to cause an infection in a wide
range of host species, including humans. Cross transmission of brucellosis can occur between cattle, swine,
sheep and goats and other species including dogs, horses, feral swine, bison, rein deer and camels [Than
(2007). Prevalence survey of bovine brucellosis (Brucella abortus) in dairy cattle in Yangon, Myanmar. A
Thesis Submitted to Chiang Mai University and Freie University at Berlin in a Partial Fulfilment of the
Requirements for the Degree of Master of Veterinary Public Health].
Keywords: Animals, Brucellosis, Diagnosis, Epidemiology, Human, Transmission, Treatment, Zoonosis
INTRODUCTION
Brucellosis is an occupational hazard with those
particularly at risk such as laboratory workers,
veterinarians, abattoir workers, farmers and animal
keepers either living in close proximity with animals
or handling aborted foetus and animal products that
are contaminated by Brucella agents (Jim, 2012). In
humans, Brucellosis is life threatening and presents
with nonspecific symptoms, including intermittent
fever, weight loss, depression, hepatomegaly and
splenomegaly. Arthritis, spondylitis, osteomyelitis,
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62
Shabu Shoukat, Henna Wani, Umer Ali, et al.
epididymitis and orchitis, as well as other more severe
complications such as neurobrucellosis, liver
abscesses and endocarditis, are common in some
patients (Bingöl et al., 1999). The risk of disease
and its severity is determined by the species of
Brucella to which an individual is exposed. Brucella
melitensis is the type most frequently reported as a
cause of human disease and the most frequently
isolated from cases. It is the most virulent type and
associated with severe acute disease. Neuro-
brucellosis in humans can occur in the form of
meningitis and meningoencephalitis. In domestic
animals, Brucella infects the reticuloendothelial
system and genital organs causing chronic infection
and abortion (especially in the last trimester), stillbirth
and infertility, which significantly affect farmers
economically due to loss of production. Epididymo-
ochitis is common in males and the females that
have aborted show necropulurent placentitis and
endometrititis. Lesions in the placenta cause oedema
of the chorionic stroma and multifocal necrosis of
allantochorion. These lesions are cardinal in the
induction of abortion and eventual infertility. This is
accompanied by large accumulation of neutrophils
and degenerate leukocytes. In addition, large number
of tiny gram-positive Brucella coccobacilli can be
seen in trophoblasts. Contrary to some traditional
views, B. melitensis remains fully virulent for man
after infecting cattle. The bovine infection presents a
particularly serious problem because of the large
volume of infected milk that can be produced by an
individual animal and because of the extensive
environmental contamination that even single
abortions or infected births can produce (Radostits
et al., 2007).
Br uce llo sis is an occupatio nal direc t
anthropozoonosis. Public health significance of
brucellosis is not only because of its status as one
of the world’s major zoonoses but also because it
causes a serious reduction in availability of quality
food, especially the animal proteins. It also poses a
serious threat to livestock economy by causing
abortion, loss of offspring, infertility and reduction in
milk yield. Most cases in humans are caused by
contact with infected animals or animal products,
such as unpasteurised milk and cheese prepared
from infected unpasteurised milk (Pappas et al.,
2005).
There are six recognised species for genus
Brucella: B. melitensis, B. abortus, B. suis, B. canis,
B. ovis and B. neotomae; and two provisional species
B. pinnipediae and B. cetaceae. This taxonomic
classification is mainly based on the difference in
host preference and pathogenicity which can be
attributed to various proteomes, as exemplified by
specific outer-membrane protein markers (Pappas
et al., 2005).
MODES OF TRANSMISSION
Both vertical and horizontal transmissions of
brucellosis exist in animals. Horizontal transmission
occurs through ingestion of contaminated feed, skin
penetration, via conjunctiva, inhalation and udder
contamination during milking or by licking the
discharge of an animal, newborn calf or retained foetal
membrane. Foetus can be infected in uterus or
suckling of infected dams. Congenital infection that
happens during parturition is frequently cleared, and
only few animals remained infected as adult. Venereal
infections can also occur and mainly seen with B.
su is inf ections. The importan ce of venere al
transmission varies with the species; it is the primary
route of transmission for B. ovis, B. suis and B. canis
are also spread frequently by this route. B. abortus
and B. melitensis can be found in semen, but venereal
transmission of these organisms is uncommon (OIE,
2009a). Of the transmission ways of brucellosis to
human, ingestion of unpasteurised dairy foods
produced from unlicenced family owned flocks whose
products are sold door-to-door at low prices is one of
the known ways. Dairy products are the main source
of infection for people who do not have direct contact
with animals. Transmission of infection to humans
occurs through breaks in the skin, following direct
contact with tissues, blood, urine, vaginal discharges,
aborted foetuses or placentas. Occupational aerosol
infection in laboratories and abattoirs has also been
documented. Accidental inoculation of live vaccines
(such as B. abortus Strain 19 and B. melitensis Rev
1) can also occur, resulting in human infections (Tigist
et al., 2011).
Most cases in humans are caused by contact
with infected animals or animal products, such as
unpasteurised milk and cheese prepared from infected
un pas teu ris ed milk (Pa ppas et al. , 200 5).
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Journal of Immunology and Immunopathology 63
Brucellosis: A Current Review Update on Zoonosis
Consumption of unpasteurised milk and their products
especially soft cheese, yoghurts and ice-cream
(Eckman, 1975), undercooked traditional delicacies
such as liver (Malik, 1997) and spleen (Elbeltagy,
2001) are mainly responsible for human brucellosis.
Camel milk is considered to be the most important
source of infection in Middle East countries and
Mongolia (Mantur and Amarnath, 2008).
MORPHOLOGY AND GROWTH REQUIREMENT
Brucellae are coccobacilli or short rods, usually
arranged singly but sometimes in pairs or small
groups. The organisms are gram negative facultative
intracellular parasites. Carbon dioxide is important
elements for growth of Brucella organism, especially
B. abortus; such organisms, which require carbon
dioxide for their growth, are called capnophilic
organisms. At pH <4, Brucella agents do not have
potential to survive.
PATHOGENESIS
Brucella may enter the host via ingestion or
inhalation, or through conjunctiva or skin abrasions.
After infecting the host, the pathogen becomes
sequestered within cells of the reticulo-endothelial
system. Organisms may be engulfed by phagocytic
cells and localised to regional lymph nodes. Then
they proliferate, disseminate haemogenously and
localise in the reticulo-endothelial and reproductive
tract. The smooth lipopolysaccharides that cover the
bacterium and proteins involved in signalling, gene
regulation and transmembrane transportation are
among the factors suspected to be involved in the
virulence of Brucella. The smooth, non-endotoxic
lipopolysaccharides help to block the development
of innate and specific immunity during the early stage
of infection; it protects the pathogen from the
microbicidal activities of the immune system and has
a role in cell entry and immune evasion of the infected
cell (Lapaque et al., 2005). The lipopolysaccharides
are thought to alter the capacity of the infected cell
to pr ese nt for eig n ant ige ns to the M ajo r
Histocompatibility Complex (class II) antigen
presentation system, hence preventing attack and
killing of the infected cell by the immune system. In
addition, smooth lipopolysaccharide in Brucella may
be involved in the inhibition of apoptosis of infected
cells, since resistance to apoptosis of infected cells
has been observed in patients with acute and chronic
disease (9). Rough strains (strains with lipopoly-
saccharide lacking the O-side chain) are less virulent
because of their inability to overcome the host
defence system; these rough strains do not confer
host cells resistance to apoptosis. From genus
Brucella, B. ovis and B. canis are classified under
rough strains group, whereas B. abo rtus, B .
melitensis and B. suis are categorised under smooth
strains group (high virulent and have O-side chain)
(Porte et al., 2003).
LABORATORY DIAGNOSIS
Bacteriology
The isolation and identification of Brucella can
confirm a diagnosis of brucellosis. Brucella is most
commonly isolated from blood cultures. It can also,
however, be isolated from the following:
bone marrow
cerebrospinal fluid
wounds
purulent discharge
joint fluid
There are a range of commercially available
culture media for growing Brucella; the most common
basal media in use are tryptcase soy, bacto tryptose,
triptic soy and tryptone soya. Frequently, field
samples are contaminated with other bacteria; thus,
selective media should be used to avoid overgrowth
by fast growing agents. Classical identification and
typing of Brucella species into their respective
species and biovars are the work that should be
undertaken after culturing any suspected specimen
in appropriate media. After 48–72 h of incubation at
37°C, Brucella colonies are 0.5 to 1.0 mm in diameter
with a convex and circular outline. Smooth strains
(B. abor tu s, B. mel itens is and B. suis ) ar e
transparent and pale yellow, resembling droplets of
honey with a shiny surface when observed in
transmitted light. Rough colonies (B. ovis and B.
canis) are more opaque with a granular surface when
compared with the smooth strains of Brucella
organisms. Dissociation of Brucella can be detected
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64
Shabu Shoukat, Henna Wani, Umer Ali, et al.
by the emulsification of a colony in 0.1 per cent w/v
aqueous acriflavine. Smooth colonies, B. abortus,
B. melitensis and B. suis, produce a yellow uniform
suspension, whereas rough colonies, B. ovis and B.
canis, produce granular agglutinates. Colonial
variation can be detected also by examining the plates
under oblique light after staining the colonies with
crystal violet. Smooth colonies appear translucent,
and pale yellow and rough colonies are stained with
red, purple or blue with opaque and granular
appearance (Fernando et al., 2010).
Serology
Serological tests can be divided broadly into two
groups, and these are screening tests lab tests.
Some screening tests used in the field clinics or in
regional laboratories are the Rose Bengal, buffered
plate agglutination test. The Rose Bengal Plate Test
(RBPT) has a very high sensitivity to ensure that
infected animals are not missed (Sharma et al.,
2017a,b; Sharma and Ganguly, 2017). The milk ring
test is also an excellent screening test for dairy
cattle. Indirect ELISA tests are also being used to
screen milk and serum. Confirmatory tests include
complement fixation tests, competitive ELISA,
fluorescence polarisation assay are very useful in
distinguishing vaccinal antibody responses from
those induced by field infections (FAO, 2003).
As far as eradication of B. melitensis from small
ruminants is concerned, no country has yet been
able to achieve it (Saleem et al., 2004). As mentioned,
the disease is present in India, in both livestock
(Renukaradhya et al., 2002; Rashid et al., 2008; Boral
et al., 2009) as well as humans (Agasthya et al.,
2007; Smith and Kadri, 2005). Moreover, >70 per cent
of Indian population is rural which is constantly
exposed to the infecte d animals resulting in
continuous transmission of disease to humans (Park,
2009). Therefore, the presence of disease in farmers,
veterinarians and other occupationally exposed groups
can never be questioned. Although, in past few
studies, involving humans and animals had been
performed in Jammu region, the status of disease in
border areas of Jammu is largely unknown; thus, the
present study was aimed to evaluate the prevalence
of brucellosis in goats and human beings along with
their epidemiological features in and around border
areas of Jammu. The study also compared the various
serological tests, namely RBPT, STAT and I-ELISA,
as they are routinely used tests in the diagnosis of
brucellosis; however, no single test in itself is fully
satisfactory, and each test is associated with certain
disadvantages (MacMillan, 1990; Nielsen, 2002;
Maansi et al., 2017). Moreover, due to the lack of
reliability of tests, the seroprevalence has been
calculated individually for each test.
Molecular Technique
Molecular biolo gy as a diagnostic tool is
advancing and will soon be at the point of replacing
actual bacterial isolation. The use of the Polymerase
Chain Reaction (PCR) to identify Brucella DNA at
genus, species and even biovar levels has become
extended to improve diagnostic tests, and a diversity
of methods have been developed. Applications for
PCR methods range from the diagnosis of the disease
to characterisation of field isolates for epidemiological
purposes including taxonomic studies. PCR-based
assays are also useful in chronically infected patients
where the yield of bacteria from blood cultures is
usually low. It is rapid, safe and cost effective, the
only real problems being some uncertainties regarding
specificity (FAO, 2003; Maansi et al., 2017).
ECONOMIC IMPORTANCE OF BRUCELLOSIS
Brucellosis is a major veterinary and human
health importance in economy of affected countries.
Among the genus B rucella, B. melitensis , B.
abortus, B. suis and B. ovis which preferentially
infect sheep and goats, cattle, pigs and sheep,
respectively, are the most important from a socio-
economic standpoint. In addition to decreasing
productivity in animals, the first three species are
the main ones responsible for brucellosis in human
beings (De Miguel et al., 2011). Costs include
production loss associated with infection in animals,
preventive programme and in human disease cost of
treatment and absenteeism from work bring many
economical impacts. Losses in animal production
due to brucellosis disease can be of major importance,
primarily because of the decreased milk production
by aborting dairy animals; the common sequel of
infertility increases the period between lactation, and
in an infected herd, the average inter calving period
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Journal of Immunology and Immunopathology 65
Brucellosis: A Current Review Update on Zoonosis
may be prolonged by several months. This is of
greatest importance in beef herds where the calves
represent the sole source of income. A high incidence
of temporary and permanent infertility results in heavy
culling of valuable, and some deaths occur as the
result of acute metritis following retention of the
placenta. The effect of the disease on ram’s fertility
can influence the number of rams that are required in
a flock; the required ram to ewe ratio is significantly
reduced in B. ovis-free flocks. The percentage of
lambs born early and within the first three weeks of
the lambing period is also markedly increased.
Disease meta-analysis showed that the highest
incidences of brucellosis occur during spring and
summer, whereas the lowest incidences during winter
and autumn seasons (Moosazadeh et al., 2016).
ZOONOTIC IMPORTANCE OF BRUCELLOSIS
Br uce llo sis is major h eal th pro blem in
Mediterranean, Middle East, India, Latin America,
Africa, parts of Mexico and parts of Asia (Sharma et
al., 2015, 2016). Syria has the highest annual
incidence worldwide with an alarming 1,603 human
cases per million per year (Boral et al., 2008, 2009).
Rate of human infection is high in Peru, Kuwait and
Saudi Arabia as compared with sub-Saharan Africa,
where the rate is relatively low, possibly due to under
reporting and low levels of surveillance (McGill, 2003).
Expansion of animal industries, the lack of
hygienic measures in animal husbandry and poor
food handling partly account for brucellosis to remain
a public health hazard. International travel and the
importation of different dairy products into Brucella
free regions contribute to the ever-increasing concern
over human brucellosis. Brucellosis is a zoonotic
disease occurring in humans and various species of
domesticated and feral animals (Sharma et al.,
2017a,b). Human brucellosis can be a very debilitating
disease, although the case fatality rate is generally
low; it often becomes s ub-clinical or chronic,
especially if not recognised early and treated
promptly. All ages of human beings are susceptible,
and even congenital cases have been recorded (FAO,
WHO and OIE, 2006). High risk groups include those
exposed through occupation in contexts where animal
infection occurs, such as slaughterhouse workers,
hunters, farmers and veterinarians (14). The three
species of Brucella of major concern here are B.
abortus (biovars 1–6), affecting primarily cattle and
other bovidae, B. suis (biovars 1–5), affecting primarily
swine; and B. melitensis (biovars 1–3), affecting
primarily sheep and goats. The persistent infection
of the mammary glands and supramammary lymph
nodes leads to a constant or intermittent shedding
of the organisms in the milk in succeeding lactations.
It provides an important source of infection for man
and young animals. Of the three species, B .
melitensis is highly pathogenic for human beings
(FAO, 2003). In addition to the above three Brucella
species, B. canis also has zoonotic importance, and
its infections in humans resemble brucellosis caused
by other Brucella species (CFSPH, 2012).
Brucellosis results in huge economic losses and
social burden on the society especially in the
developing countries, and it is one of the most
common occupational anthropozoonoses present
world-wide. Due to the slow onset and absence of
pathognomonic signs and symptoms, it is usually
diagnosed quite late and by the time the animal herd
is already infected. Important animal species of genus
Brucella, namely B. abortus associated with cattle
and B. melitensis associated with sheep and goat
(Corbel, 1997). In humans, 90 per cent cases of
brucellosis are said to be associated with B.
melitensis (Benkirane, 2006).
Prevalence of brucellosis poses a direct public
health implications and act as a potential barrier to
international trade of animals and animal products
(WHO, 1997). Some recent reports regarding
Brucella infection involving marine mammal and birds
have given a new orientation to the epidemiology of
brucellosis. Although dairy cattle, sheep, goats and
swine are major reservoirs of the disease and horses,
camels and wild animals can also suffer occasionally.
Brucellosis management remained a difficult task as
control programmes require vaccination of animals
besides other drastic measures. However, the
difficulty in differentiating between vaccinated and
infected animals has posed a serious limitation in
interpretation of results during vaccination campaigns
(OIE, 2008a,b, 2009b,c).
Although the disease is eradicated in bovines
(B. abortus), in many developed countries like
Australia, Canada, Cyprus, Denmark, Finland,
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Vol. 19, No. 2, July-December, 2017
66
Shabu Shoukat, Henna Wani, Umer Ali, et al.
Netherlands, New Zealand, Norway, Sweden and
United Kingdom, it is still present in Mediterranean
countries of Europe, Northern and Eastern Africa, near
East countries, India, Central Asia, Mexico, Central
and South America (Seleem et al., 2010). As reported
by Schwabe (1984), in India, there have been annual
losses of Rs. 240 millions due to bovine brucellosis
alone.
As far as eradication of B. melitensis from small
ruminants is concerned, no country has yet been
able to achieve it (Saleem et al., 2004). As mentioned,
the disease is present in India, in both livestock
(Renukaradhya et al., 2002; Rashid et al., 2008; Boral
et al., 2009) as well as humans (Agasthya et al.,
2007; Smith and Kadri, 2005). Moreover, >70 per cent
of Indian population is rural which is constantly
exposed to the infecte d animals resulting in
continuous transmission of disease to humans (Park,
2009). Therefore, the presence of disease in farmers,
veterinarians and other occupationally exposed groups
can never be questioned. Although the past few
studies, involving humans and animals, had been
performed in Jammu region, the status of disease in
border areas of Jammu is largely unknown; thus, the
present study was aimed to evaluate the prevalence
of brucellosis in goats and human beings along with
their epidemiological features in and around border
areas of Jammu. The study also compared the various
serological tests, namely RBPT, STAT and I-ELISA,
as they are routinely used tests in the diagnosis of
brucellosis; however, no single test in itself is fully
satisfactory, and each test is associated with certain
disadvantages (MacMillan, 1990; Nielsen, 2002;
Sharma et al., 2017a,b; Maansi et al., 2017; Sharma
and Ganguly, 2017). Moreover, due to the lack of
reliability of tests, the seroprevalence has been
calculated individually for each test.
CONCLUSION
Brucellosis is a major bacterial zoonosis of
global, economic and public health significance
(Cutler and Whatmore, 2003). The disease has also
been reported in recent years from wild and marine
mammals and birds. The presence of brucellosis in
wild animals with a potential for continuous transfer
to domestic animals and from them to humans is
another epidemiological issue (Cutler et al., 2005).
In cattle, B. abortus causes abortion, stillbirth and
weak calves with abortions usually occurring during
the third trimester of gestation. In goats, B. melitensis
can cause abortion, retained placenta, orchitis and
epididymitis. Abortions usually occur during the fourth
month of gestation in goats. Clinical manifestations
of disease in man are weakness, fever, profuse
sweating e sp ec ia lly in night, los s of weight,
generalised body ache and others. Swelling in testes
and burning micturition due to orchitis and urethritis,
respectively, are also peculiar symptoms of disease
in man (Smith and Kadri, 2005).
Brucellosis in India is very common but often a
neglected disease (Renukaradhya et al., 2002).
Brucellosis due to B. melitensis is widespread in India
and major cause of abortion in small ruminants
imposing economic loss due to an adverse effect on
total animal protein supplies and severe hazard to
human health (Abeer et al., 2003). Sheep brucellosis
can be divided into classical brucellosis and ram
epididymitis. Ram epididymitis is caused by non-
zoonotic agent B. ovis, whereas classical brucellosis
constitutes a major public health threat equal to goat
brucellosis (Acha and Szyfres, 2003). There are about
500,000 new human cases of brucellosis reported
annually worldwide making it most common zoonosis
(Saleem et al., 2004; Seleem et al., 2010).
Food safety is one of the principal pillars on which
protection of human health resides; hence humans
ar e i nfe cted by Bru cel la ma inly throu gh
inappropriately prepared and/or preserved food of
animal origin. Laws, regulations and veterinary policy
measures alone will not bring the desired results.
The whole community needs to be involved through
health education in schools, in the workplace and in
the population at large (FAO, 2003). Brucellosis is
the most common zoonosis worldwide with over
500,000 cases every year. In general, prevention of
brucellosis like any other zoonoses begins by
elimination of the pathogens in animals. The aim of
animal brucellosis control programme is to reduce
the impact of a disease on human health and the
economic consequences. The elimination of the
disease from the population is not the objective of a
control programme, and it is implicit that some
‘acceptable level’ of infection will remain in the
population. Control programmes have an indefinite
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Journal of Immunology and Immunopathology 67
Brucellosis: A Current Review Update on Zoonosis
duration and will need to be maintained even after
the ‘acceptable level’ of infection has been reached,
so that the disease does not re-emerge.
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