Obstetrics and Genital

Diseases
4(2-2)
Lec : Bovine Abortion: Infectious causes and Brucellosis
 Infectious causes
Bacterial:
Brucellosis
Brucella abortus

Leptospirosis
Leptospira pomona
Leptospira icterohemorrhagica

Campylobacteriosis
Campylobacter fetus venerealis

Listeriosis
Listeria monocytogenes

Tuberculosis
Mycobacterium bovis
 Infectious causes
• Miscellaneous bacterial:
Contd…..
Streptococcus spp. Staphylococcus spp.

Escerichia coli Pseudomonas aeruginosa

Corynebacterium pyogenes
Campylobacter fetus intestinalis

Mycobacterium avium Pasteurella multocida

Salmonella paratyphi B Salmonella dublin

Bacillus anthracis Mycoplasma spp.

Hemophilus spp. Alcaligenes fecalis

Erysipelothrix insidiosa
 Infectious causes contd…..
VIRAL:

IBR-IPV
(Infectious Bovine Rhinotracheatis and Infectious
Pustular Vulvovaginitis)

Epizootic Bovine Abortion

 Infectious causes contd…..
• Miscellaneous Viral:
– FMD (Foot and Mouth disease virus)
– RP (Rinderpest)
– RVF (Rift valley fever)
– Bovine infectious petechial fever
– Tick borne fever
– BVD-MD (Bovine viral diarrhea-Mucosal Disease)
– MCF (Malignant catarrhal fever)
– Myxovirus parainfluenza-3
 Infectious causes contd…..
• Mycotic/Fungal:
– Aspergillus spp.
– Mucorales spp.
• Absidia
• Mucor
• Rhizopus
• Yeasts
 Infectious causes contd…..
• Protozoa:
– Trichomoniasis
– Toxoplasmosis
– Besnoitis/Globidiosis
– Trypanosomiasis
– Anaplasmosis
– Babesiosis
– Piroplasmosis
 Introduction
• Ch. by abortion, infection of uterus and fetal membranes
• Small, G –ve, rod
• Due to protective outer protein coat, it grows
Intracellularly and can grow in host macrophages and
also capable of chronic infection
• 1887 David Bruce - first isolated this organism (Brucella
melitenses) from spleen of patient died of Mediterranean
fever
• 1897 Fredrick Bang – isolated similar org from bovine
aborted fetus and call it as Bacillus abortus (also called
Bang’s disease)
• 1914 – Br suis was isolated from aborted porcine fetus
 Transmission
Entry into the herd of:
• Usually infected cow
• less commonly infected bulls
• rarely horse with fistulous withers or poll evil
Within the herd
• Ingestion of infected material (fodder and drinking water
become contaminated with aborted material, vaginal
discharge, etc.)
• Via respiratory tract
• Conjunctiva
• Broken/ even unbroken skin
• Tick bites
Fistulous Withers (Horse)
Poll Evil (Horse)
 Transmission
 Aborted fetuses
 Fetal membranes
 Vaginal discharges
 Milk, Colostrum, feces all material contain Brucella. These
contaminate environment including Pastures and water
supply.

• Recovered cows (not aborting) continue to excrete Brucella in

milk for many years
• Infected bulls unlikely to spread through coitus but may
transmit through AI ???
• These bulls can spread through contamination of environment
by feces and urine
 Pathogenesis
• Infection (ingestion), bacteremia then
• Usually localization of bacteria in reticulo-endothelial
tissue & reproductive organs
• Less frequently localization in bones & joints
• NP = remain localized but if preg then…..
• Production of carbohydrate erythritol by fetoplacental
unit (placentome) favors the multiplication of Brucella
(….villi….)
• Bovine, sheep, goats reproductive lesions mostly
cause death of fetus along with abortion …..excretion
continue for 2-3 wks
• Zoonotic (real occupational hazard for vets, AIT,
slaughtermen and farmers)
 Prevalence/ incidence

• Worldwide

• In Pak, 1981-87, 6416 buff/cow samples

tested serologically. 59 positive (0.9 %)
 Age buffalo cow camel Total

No. +ve No. +ve No. +ve No. +ve

% % % %
Upto 1 66 ---- 16 ----- ---- ----- 82 -----
yr
1-7 yr 140 3.57 78 1.28 12 ----- 230 2.61

> 7 yrs 547 3.47 175 2.86 69 2.90 791 3.29

Total 753 3.19 269 2.23 81 2.47 1103 2.90

• Abubakar et al,. 2010 serologically tested
at village level
< 2 yrs = 3.57 2.98 cattle / buff
>2 yrs = 6.54 12.50 cattle / buff
• 1987-92 LES, Qadirabad Xbred 6 to 12 %
• 2003-04 Bhunikey (Now BRI), >15 % in
pure exotic cows
 Etiology
Host Main Pathogen isolated Other pathogen may
involve
Cows/Buffaloes Brucella abortus Brucella melitensis, Brucella
suis
Sheep Br melitensis, Br ovis Br abortus

Goat Br melitensis Br abortus

Horse Br abortus Br suis

Pig Br suis Br melitensis, Br abortus, Br

neotomae

Dog Br canis Br abortus, Br melitensis, Br

suis
Human Br abortus, Br Br canis
melitensis, Br suis
 Economic losses
• Abortion
• Loss of fetus
• Less milk
• Male sterility
• Zoonotic (Public health hazard)
 Symptoms
• Bovines Incub period: highly variable & is inversely proportional to
stage of pregnancy at the time of infection
• Bacteremia observed 14 days after 1st sign
• Abortion occur 24-72 hrs after fetal death
• Abortion after 5th month, with the relaxation of sacroisciatic lig and
followed by brown discharge, retention of placenta, metritis leading to
infertility or sterility
• Nomads, this discharge is the only sign observed and fetus and
membranes mostly not seen
• Org spreads rapidly leading to abortion storm
• Mostly carry subsequent calves to full term. Sometimes 2nd or even
3rd time abortion in the same individual
• Stage of herd resistance only heifers carrying 1st calves will abort.
 Symptoms
• Calves born from contaminated uteri or become infected after
birth, don’t retain infection but only 5 % may retain infection for life
remaining in dormant carrier state until sexually mature and
infection flares up.
• Bulls mostly no clinical signs, rarely orchitis, reduced libido,
infertility
• Horses fistulous withers, poll evil, fever, lameness due to arthritis/
laminitis
• Sheep/goats abortion 3rd / 4th month or still birth, lameness,
mastitis (discoloration of milk with clots) but in acute, weakness,
fever, diarrhea
• Dogs EEM or abortion at 50th day but male may suffer scrotal
dermatitis, unilateral test atrophy
 Symptoms
• Man milkmen, vets have more chance to
contact the disease (through skin, removal
retained placenta, drinking of milk from
Brucella affected cows/buff)
• Also called Malta fever
• ch. by intermittent chills & fever or undulant
fever, pain in joints and muscles, orchitis,
mental depression, general sickness.
• Chronic form in man debilitating
 Lesions
• Bovines Lymph nodes, udder, uterus, joints, liver,
spleen mostly involved
• Cotyledons color changes from normal bright red to
dirty greyish yellow
• necrotic placentitis, swollen and hyperemic
cotyledon surrounded by brownish exudate
• Intercotyledonary spaces thick with leathery
appearance
• Infected udder rarely show heavy cellular infiltration,
degeneration, necrosis
 Lesions
• Bulls orchitis, abscess/ necrotic
areas in testicles surrounded by
fibrous CT
• Sheep / goats in female, same as
in bovine but in rams epididymal
tail and seminal vesicles are
involved
 Diagnostic techniques
• Bacteriological methods

• Serological methods

• Allergic tests
 Bacteriological methods
Samples to be collected
– Milk & milk products, blood, semen
samples
– Vaginal secretion
– Aborted material
• Placenta
• Aborted fetus (stomach contents, lungs, spleen,
muconium, testes of male fetus)
 Bact…..Methods
Isolation of Brucella
• Culture Media
– Serum dextrose Agar
– Glycerol dextrose agar
– Potato infusion agar
– Serum potato infusion agar
• Animal inoculation method
Demonstration by staining
Smears from vaginal swabs, stomach contents of aborted
fetus and placenta can be made with modified Zeihl-
Neilson method
 Serological methods
• SAT
• RBPT
• CFT
• Competitive ELISA
Other than serum
• MRT
• WAT (which is less sensitive than MRT)
 MRT
• It tells positivity even in pooled milk sample, e.g., 5-10 negative
mixed with 1 positive
• 1 ml milk in narrow tube + add 1 drop (0.03 ml) of MRT Ag, mix
by inverting and avoid frothing, incubate 1 hr at 37 C
Creamy Ring Milk Column Rating
Deeply violet colored white ++++
Less color slight color +++
Less color moderate color ++
Same Color Same Color +
White deep violet color -

Note: False +ve from individual cows may be just after

parturition or at end of lactation
 RBPT
• Result agree very closely with CFT
• RBPT identifies infected animals earlier than any other especially
SAT
• When incidence is low and calfhood vaccination is widely used,
its results are over sensitive
• Mostly after RBPT, other test SAT etc are recommended to
overcome over sensitivity
• 1 drop of Ag & 1 drop of test serum on enamel plate, side by side
• Mix with wooden sterilized toothpick
• Agglutination occur in 0 – 4 minutes
• Read the agglutination in good light
• It is good to observe when mixture flows away from operator
 Serum Agglutination Test (SAT)
• 5 conical tubes
• 0.8 ml NS (containing 0.5 % phenol) in 1st tube
• 0.5 ml of same NS in other four tubes
• 0.2 ml of test serum to 1st tube, mix, 1:5 dilution
• 0.5 ml withdrawn from 1st and carried to 2nd , mix, 0.5 ml withdrawn
from 2nd and carried to 3rd and so on
• Now dilution in all 5 tubes 1:5, 1:10, 1:20, 1:40, 1:80
• 0.5 ml of Br abortus Ag to each tube with final dilution of test serum
to 1/10, 1/20, 1/40, 1/80, 1/160
• Mix from 1/160 dilution to downward
• Known +ve and known –ve control
• Incubate for 37 C for 24 hrs
 SAT-Interpretation of results
• Based upon clearance of suspension along with sedimentation
• ++++ recording = clearance of suspension is100 % with marked
sediments
• +++ recording = clearance of suspension is 75 % with marked sediments
• ++ recording = clearance of suspension is 50 % with marked sediments
• + recording = clearance of suspension is 25 % with marked sediments
Interpretation
1/10 1/20 1/40 1/80 1/160
+ + + - - Negative
+ + + + + Negative
++ + + + + Doubtful
+++ ++ + + + Doubtful
++++ +++ ++ + + Positive
 Allergic test
Brucelhydrolysate (Sheep/goats)
1st day 0.2 ml is injected i.d. into the caudal
fold.
Results are read at 48 hours, a positive
sample is characterized by
• Hyperemic
• Edematous swelling of skin fold
All non-reacting animals are inoculated again
on the 3rd day, near the site of the 1st
injection and results are read after 24 hours.
 Control
Two methods for control:
1. Vaccination
2. test and slaughter

•After infection or vaccination, both cell

mediated and humoral immune responses
are stimulated although former is more
important
 vaccination
Brucella abortus Strain 19 vaccine
• Live attenuated, Single calfhood vaccination at the
age 3-6 months of age, sufficient immunity for 5 or
more lactations
• In adult, not so much effective
• Efficacy 65-75 %
• It reduces the rate of abortion in herd but cannot
prevent mastitis
• Less virulent but it may result in abortion if given
during late pregnancy
Brucella abortus Strain 19 vaccine
• Calves just after birth heavy exposure to infection
refractory to vacc
• In males, of no value rather may result in orchitis
seminal vesiculitis and started shedding org in
semen
• Anaphylactic reaction may occur
• In lactating, causes high but temporary drop in milk
production
• Accidental splashing or pricking with vaccine needle
during vaccination may cause undulant fever in man
 Brucella abortus strain 45/20 vaccine

• May provide immediate protection if

required in cattle

• In adult cattle, 2 doses 6-12 weeks apart

• Very less effective when used as calfhood

vaccination
 Some more strategies for control
Rules to follow when adjacent to infected herd
• Don’t allow contact with neighboring stock, double
fence may be done
• Don’t share neighbor’s bull
• Segregate animals in groups esp of late pregnancy
• Use Br strain 19 or 45/20 vaccine
• Keep calves in isolation, blood test of calves 14 days
after calving, then put in main herd
• Keep in mind the danger of spread through transport
vehicles
 Treatment
• Intracellular, so protected from host defenses
and activity of antibiotics
• Incurable
• But 15 % self recovery
• Sulpha, streptomycin, tetracycline,
chloramphenicol have been tried with very
little success
• Intracellular so prolonged treatment is required
just like TB
Rules to follow to buy replacements in
herd
• don’t buy pregnant heifers or adult
• Buy fully accredited, nonpregnant
heifers, blood test 30 days after arrival
• Keep them separate from rest of herd
until clear test 14 day after calving
 Rules to follow when herd is already infected

• Regard the remainder as infected if one animal

in a group aborts
• Blood test , isolate the reactors to other
premises, cull them
• Calving pens for isolation
• Vaccinate adult non-pregnant stock
• Dispose off aborted calves, placentae, bedding,
etc. by deep burying or incineration
• Disinfect area of abortion
Preventive measures during disease

• Infected animals be isolated from the

others
• Aborted fetuses, placentae, bedding,
etc. should be buried deep or
incinerated
• Abortion area should be properly
disinfected