Brucellosis in Organized Dairy Farm: An Investigation
Amit Kumar Verma,
Brucellosis is an important zoonotic disease of animals and man causing different
reproductive problems like abortion, retention of placenta, repeat breeding,
still birth and many more. In this report, authors described an investigation
of brucellosis outbreak in an organized dairy farm, Agra, India causing abortions
and other reproductive disorders. Blood and environmental samples including
feed and water samples were collected in sterile containers. About 80% sera
samples collected from aborted animals and 60% sera samples from pregnant and
in-contact animals, maintained at the dairy farm contained antibodies against
Brucella spp., while all the bulls were found negative for presence of
anti-Brucella antibodies. Escherichia coli and Streptococcus
spp., isolates were recovered from water samples, while Staphylococcus,
Streptococcus spp. and Pseudomonas spp. could be isolated from feed
samples. In the present investigation, high prevalence of anti-Brucella
antibodies was found in sera samples of the animals, indicates that they all
had an exposure to Brucella spp. which might be the probable reason of
abortion storm. Thus, on the basis of our findings, it can be deduced that Brucella
might be associated with abortion in dairy animals even without apparent septicaemia
or any other manifestations. The presence of anti-Brucella antibodies
in sera samples of animals can be used as an indicator of infection in farm
Received: October 23, 2013;
Accepted: November 08, 2013;
Published: February 27, 2014
Reproductive disorders particularly abortion and repeat breeding in animals
are caused by various etiological agents such as Brucella spp., Infectious
Bovine Rhinotrachitis, Bovine viral diarrhoea virus, Leptospira and
Listeria spp., Campylobacter spp. etc (Refai,
2002; Renukaradhya et al., 2002; Chahota
et al., 2003; Smits and Kadri, 2005; Kumar
et al., 2009; Dhama et al., 2013;
Rathore et al., 2013; Verma
et al., 2014). Besides these some other bacteria like Staphylococcus,
Escherichia coli and other pathogenic bacteria and protozoa like Neospora
may also cause abortion in the pregnant cattle either alone or in concert with
other pathogens (Van Wuijckhuise and Otten, 2008; Njiro
et al., 2011; Blumer et al., 2011).
Among all these, Brucella abortus is considered as the most common cause
of abortion and subsequently considerable economic losses to dairy industry.
Brucellosis was first time reported in India in Anonymous
(1918) and since then it has been reported in many states of the country
and its incidence and prevalence is increasing day by day due to increase in
trade and movement of livestock (Renukaradhya et al.,
2002; Kumar et al., 2009). According to
an estimate, the annual losses due to brucellosis are Rs 350 million per year
in India (Tiwari et al., 2013). Contaminated
feed and water, close contact, unhygienic management practices are some of the
important sources of infection for terrestrial animals (PD-ADMAS,
2011). Brucella can survive and remain infective for several months
in water, aborted foetus and foetal membranes, faeces and liquid manure, organic
material, dust and soil on buildings, equipment and clothes (Alton,
1985; FAO/WHO, 1986). This article deals with an
outbreak of brucellosis in an organized dairy farm, Agra, Uttar Pradesh, India
as well as its transmission.
MATERIALS AND METHODS
Study area: The incident occurred in an organized dairy farm at Agra,
Uttar Pradesh, India. Climatic conditions of this area is humid subtropical
with dry winter type, annual rainfall (650-1000 mm), relative humidity (20-50%),
annual temperature (11.0 to 36.9°C).
Animal and management: The animal population at the farm during the
incident was 806 of different age and physiological status (Table
1). The herd were kept in groups according to their physiological stage.
The animals are kept on stall feeding and fed with wheat straw along with concentrate
and mineral mixture. They had free access to water sources (water toughs). Animals
were vaccinated against pasteurellosis with Pasteurella multocida biotype
A vaccine (Biological Product Section, Badshabagh, Lucknow) and against foot-and-mouth
disease. The hygiene and sanitation conditions at the farm were very poor or
unsatisfactory (Fig. 1a-d).
History, clinical examination and data collection: Abortions started
in the month of March, 2013 and going on with an average of 5-10 abortions per
month in dairy farm. All the animals were clinically examined by the experts
from the university. The health and basic record books of the herd, compiled
by veterinary and animal care staff, were also examined and analysed for occurrence
of the disease, morbidity and mortality etc.
Sample collection: Besides, blood samples from aborted, pregnant animals
and from bulls were also collected aseptically and transported on ice to the
laboratory. Feed and water samples were analysed for presence of contamination.
Sera from all the blood samples were separated and stored at 20°C, while
clots were processed for the isolation of bacteria. Environmental samples from
the dairy farm including water (from water troughs) and feed samples were collected
and brought in sterile containers on ice.
|| Distribution of animals according to species and physiological
|| Poor hygienic conditions at the farm
Laboratory examination: Sera samples were tested for presence of anti-Brucella
antibodies using Rose Bengal Plate Test as per the method described by Alton
et al. (1975). Briefly, the test sera and Rose-Bengal antigen were
kept for one hour at room temperature before the beginning of the test. A result
was considered positive when there was any degree of agglutination noticeable
and the absence of agglutination was considered as negative. Blood clots were
processed for bacteriological examination by incubating the samples at 37°C
onto nutrient agar, MacConkey agar and 5% sheep blood agar plates and examined
after 24-48 h. Environmental samples from the dairy farm including water and
feed samples were processed for microbiological examination as per the method
described by Cruickshank et al. (1975).
RESULTS AND DISCUSSION
A total of 16 sera samples from aborted animals (5), pregnant animals (5) and
bulls (6) were processed for diagnosis of Brucella infection using RBPT.
Out of these, 80.00% of aborted animals (4/5) and 60% of pregnant animals (3/5)
were positive for brucellosis infection, while all the bulls were found negative.
Escherichia coli and Streptococcus spp. isolates were recovered
from water samples, while Staphylococcus spp., Streptococcus spp.
and Pseudomonas spp. could be isolated from feed samples.
Reproductive-related problems in animals may be due to various infectious diseases
of bacterial origin (brucellosis, salmonellosis, campylobacteriosis, leptospirosis,
mycoplasmosis and listeriosis etc) and viral origin (Infectious Bovine Rhinotrachitis
and Bovine herpes virus-1 (BHV-1) leading to major threats to dairy industry
(Chahota et al., 2003; Kumar
et al., 2009; Njiro et al., 2011;
Murray, 2012; Watson et al.,
2012; Verma et al., 2014). Although, a number
of different bacteria have been associated with abortion in dairy animals. In
the present investigation, high prevalence of anti-Brucella antibodies
was found in sera samples of the animals, indicates that they all had an exposure
to Brucella spp. and which might be the probable reason of abortion storm.
This observation is in concurrence with earlier study (Chahota
et al., 2003) revealing involvement of Brucella spp. in all
affected cows using serological tests.
In the present study, water available to animals was heavily contaminated (105
CFU mL-1) particularly with faeces such as faecal coliforms as count
was much higher (80/100 mL-1) than the maximum permissible limits
(10/100 mL-1) in raw drinking water of livestock of dairy receiving
disinfection only (Warrington, 2001). It seems that heavily
contaminated water might also helped in transmission of infection to farm animals,
eventually leading to abortions. Treatment of affected animals for brucellosis
is not cost effective so it can be controlled by adopting the prevention and
control strategies including screening of animals, segregation of positive animals
and vaccination of healthy animals etc (OIE, 2008).
From this investigation, it can be deduced that Brucella might be associated
with abortion in dairy animals even without apparent septicaemia or any other
manifestations. The presence of anti-Brucella antibodies in sera samples
of animals can be used as an indicator of infection in farm animals. Being a
zoonotic disease, all the protective and preventive measures should be applied
for its control in animals as well as human beings. On the basis of above facts
along with availability of suitable vaccine and diagnostics, a control programme
for this disease should be started as early as possible.
Authors are highly thankful to Dean, College of Veterinary Science and Animal
Husbandry and Honble Vice Chancellor,
Uttar Pradesh Pandit Deen Dayal Upadhayay Pashu Chikitsa Vigyan Vishwavidyalay
Evum Go-Anusandhan Sansthan (DUVASU), Mathura, India; for providing all the
necessary support and facilities for conducting this study. Authors wish to
thank staff of Department of Veterinary Epidemiology and Preventive Medicine
and Department of Veterinary Public Health for their technical assistance.
1: Alton, G.G., L.M. Jones and D.E. Piezt, 1975. Laboratory Techniques in Brucellosis. 2nd Edn., WHO, Geneva, Switzerland, pp: 112-113.
2: Alton, G.G., 1985. The Epidemiology of Brucella Melitensis in Sheep and Goats. In: Brucella Melitensis: A Seminar in the CEC Programme of Co-ordination of Research on Animal Pathology, Verger, J.M. and M. Plommet (Eds.). Martinus Nijhoff, Dordrecht-Boston-Lancaster, pp: 187-196.
3: Anonymous, 1918. Annual report 1917-1918. Imperial Institute of Veterinary Research, Mukteswar, Uttar Pradesh, India.
4: Blumer, S., G. Greub, A. Waldvogel, M. Hassig and R. Thoma et al., 2011. Waddlia, parachlamydia and chlamydiaceae in bovine abortion. Vet. Microbiol., 152: 385-393.
CrossRef | PubMed | Direct Link |
5: Chahota, R., M. Sharma, R.C. Katoch, S. Verma, M.M. Singh, V. Kapoor and R.K. Asrani, 2003. Brucellosis outbreak in an organized dairy farm involving cows and in contact human beings, in Himachal Pradesh, India. Vet. Arhiv, 73: 95-102.
Direct Link |
6: Cruickshank, R., J.P. Duguid, B.P. Marmion and R.H.A. Swain, 1975. Medical Microbiology: The Practice of Medical Microbiology. 12th Edn., Churchill Livingstone, Edinburgh, UK., ISBN-13: 9780443011115, Pages: 587.
7: Dhama, K., A.K. Verma, R. Tiwari, S. Chakraborty and K. Vora et al., 2013. A perspective on applications of Geographical Information System (GIS): An advanced tracking tool for disease surveillance and monitoring in veterinary epidemiology. Adv. Anim. Vet. Sci., 1: 14-24.
Direct Link |
8: FAO/WHO, 1986. Joint FAO/WHO expert committee on Brucellosis. 6th Report, Technical Report Series 740, WHO, Geneva, Switzerland. http://libdoc.who.int/trs/WHO_TRS_740.pdf.
9: Kumar, N., B.C. Pal, S.K. Yadav, A.K. Verma, U. Jain and G. Yadav, 2009. Prevalence of bovine brucellosis in Uttar Pradesh, India. J. Vet. Public Health, 7: 129-131.
Direct Link |
10: Murray, R.D., 2012. Laboratory diagnosis of Mycoplasma/Ureaplasma abortion in cattle. Vet. Record, 170: 130-131.
CrossRef | Direct Link |
11: Njiro, S.M., A.G. Kidanemariam, A.M. Tsotetsi, T.C. Katsande and M. Mnisi et al., 2011. A study of some infectious causes of reproductive disorders in cattle owned by resource-poor farmers in Gauteng province, South Africa. J. S. Afr. Vet. Assoc., 82: 213-218.
12: OIE, 2008. Bovine Brucellosis: Manual of Diagnostic Tests and Vaccines for Terrestrial Animals (Mammals, Birds and Bees). Chapter 2.4.3, 6th Edn., Vol. 2, OIE, Paris, France, pp: 624-659.
13: PD-ADMAS, 2011. Animal disease monitoring and surveilance report 1994-2002. IAH&VB, Bangalore, India.
14: Rathore, R.S., A.K. Verma, S. Rajagunalan and K. Dhama, 2013. Trends in Campylobacter Diagnosis. In: Veterinary Diagnostics: Current Trends, Gupta, R.P., S.R. Garg, V. Nehra and D. Lather (Eds.). Chapter 19, Satish Serial Publishing House, Delhi, ISBN: 978-93-81226-49-0, pp: 321-339.
15: Refai, M., 2002. Incidence and control of brucellosis in the Near East region. Vet. Microbiol., 90: 81-110.
CrossRef | Direct Link |
16: Renukaradhya, G.J., S. Isloor and M. Rajasekhar, 2002. Epidemiology, zoonotic aspects, vaccination and control/eradication of brucellosis in India. Vet. Microbiol., 90: 183-195.
CrossRef | Direct Link |
17: Smits, H.L. and S.M. Kadri, 2005. Brucellosis in India: A deceptive infectious disease. Indian J. Med. Res., 122: 375-384.
PubMed | Direct Link |
18: Tiwari, R., M.C. Sharma, K.K. Mishra and B.P. Singh, 2013. Economic impacts of infectious diseases of livestock. Indian J. Anim. Sci., 83: 316-320.
Direct Link |
19: Van Wuijckhuise, L. and J. Otten, 2008. Abortions, fertility problems and mastitis on a cattle farm and a sick farmer: A zoonosis? Infection with Staphylococcus aureus phagetype T543. Tijdschr Diergeneeskd, 133: 944-946.
PubMed | Direct Link |
20: Verma, A.K., A. Kumar, Sahzad, N.C.R. Reddy and A.N. Shinde, 2014. Sero-prevalence of infectious bovine rhinotracheitis in dairy animals with reproductive disorders in Uttar Pradesh, India. Pak. J. Biol. Sci. 10.3923/pjbs/2014
21: Warrington, P.D., 2001. Water quality criteria for microbiological indicators. Pursuant to Section 2(e) of the Environment Management Act, 1981, pp: 2. http://www.env.gov.bc.ca/wat/wq/BCguidelines/microbiology/microbiology.html.
22: Watson, P., C. Mason, H. Stevenson, S. Scholes and A. Schock et al., 2012. Laboratory diagnosis of Mycoplasma/Ureaplasma abortion in cattle. Vet. Record, 170: 82-84.