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Sri Lanka
Veterinary
Journal
Volume 63 (2) July - December 2016
e O cial Journal of the Sri Lanka Veterinary Association
SRI LANKA
VETERINARY JOURNAL
THE OFFICIAL JOURNAL
OF THE SRI LANKA VETERINARY ASSOCIATION
275/75, OPA Building
Prof. Stanley Wijesundara Mawatha
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Vol. 63 (2)
JULY -DECEMBER 2016
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sent to the Editorial Secretary, Sri Lanka Veterinary Journal
Dept. of Biochemistry, Faculty of Medicine, Peradeniya, Sri Lanka
SRI LANKA VETERINARY JOURNAL
VOL. 63 (2)
July - December 2016
EDITORIAL COMMITTEE
R. SIVAKANESAN
B.V.Sc (Ceylon), Ph.D (Hull), Editor
INDIRA D. SILVA
MANGALA GUNATILAKE
MISSAKA P. B. WIJAYAGUNAWARDANE
D.D. NIRANJALA DE SILVA
S. S. P. SILVA
P. A. BASIL. D. ALEXANDER
R.M.C. DESHAPRIYA
L. G. SAMPATH LOKUGALAPPATTI
CHANAKA RABEL
ERANDA RAJAPAKSHA
BVSc (Sri Lanka), PhD (Davis, California) Co-editor
BVSc (Sri Lanka), PhD (Sri Lanka)
BVSC (Sri Lanka), PhD (Japan)
BVSc (Sri Lanka), PhD (Cantab)
BVSc (Sri Lanka), PhD (UK)
BVSc (Sri Lanka), PhD (Guelph)
BVSc (Sri Lanka), MSc (Pera), MPhil (UK), PhD (Japan)
BVSc (Sri Lanka), MSc (Sri Lanka) PhD (Stellenbosch)
BVSc (Sri Lanka) MS (Illinois), PhD (Illinois)
BVSc (Sri Lanka) MSc (Edinburgh),
PhD, Dip. Vet. Beh (Davis, California)
BVSc (Sri Lanka), PhD (Sri Lanka) Secretary
D. R. ANURUDDHIKA DISSANAYAKE
CONTENTS
SECTION A
COMPLEX CLINICAL PRESENTATIONS OF TICK-BORNE DISEASES IN DOGS IN SRI LANKA
Indira Silva
1
ASSESSMENT AND COMPARISON OF URINARY PROTEIN: CREATININE RATIOS OF HEALTHY
DOMESTIC DOGS AND WORKING DOGS IN KENNELS DIVISION IN SRI LANKA POLICE FOR
EARLY DETECTION OF RENAL DISEASE
K.N.A. Wijayawardhane, K.G.S.S. Karunathilake and V.S.S. Kumara
11
BODY CONDITION SCORE IN LARGE PURE BRED DOGS: A PRELIMINARY STUDY ON AGREEMENT
BETWEEN OWNER'S PERCEPTION AND SCIENTIFIC EVALUATION
M. Jagatheesan, D. D. N De. Silva and H. M. H. S . Ariyarathna
17
FELINE PANLEUKOPENIAVIRUS INFECTION IN ACAPTIVE-BRED BENGAL TIGER (Panther tigris tigris)
AND A LEOPARD (Panthera pradus)
D.R.A. Dissanayake, I.D. Silva, Sashikala Gamage, Dinuka Sonnadara, M.R.B.N. Bandara,
S.S. Alokabandara, V.P. Priyanka Jayapani and Waruni Jayaweera
23
PREVALENCE, TREATMENT AND MANAGEMENT OF HEALTH CONDITIONS IN PET RABBITS
PRESENTED TO THE VETERINARY TEACHING HOSPITAL (VTH), UNIVERSITYOF PERADENIYA
K.K. Sumanasekera, M.G.C.M. Jayasinghe, K.G. Indika, R.E.H. Perera and D.D.N. de Silva
27
SECTION B
FOOD SAFETY MONITORING AND S URVEILLANCE SYSTEM FOR SRI LANKA
Umanga C. Gunasekera, Mahalingam Muralithas and Maximilian P.O. Baumann
31
HUMAN-MONKEY (Macaca sinica) CONFLICT IN SRI LANKA
B.C.G. Mendis and Ashoka Dangolla
35
LETTER TO THE EDITOR
38
ABSTRACTS
10, 22
S.L.Vet.J. 2016, 63:2(A): 1-9
Review article
COMPLEX CLINICAL PRESENTATIONS OF TICK-BORNE DISEASES
IN DOGS IN SRI LANKA
Indira Silva
Veterinary Teaching Hospital, University of Peradeniya, Sri Lanka
Vector-borne diseases (VBD) which were mostly seen
in the tropics, in locations with limited resources and
surveillance (Gubler, 2009), have now reached high or
medium occurrence in all continents, except Australia
[CVBD (Companion Vector Borne Diseases) website].
A global re-emergence of VBDs, with a high frequency
of transmission has been recorded due to multiple
environmental and social factors, such as, increased
human and animal bonding, mobility across borders,
animal trade, global warming, resistance to pesticides,
changes in agricultural practices, deforestation and
reforestation and loss of biodiversity (WHO, Vectorborne diseases, February 2016). Pets are not considered a
direct source of infection to people, but direct
transmission from an infected animal can occur via
accidental inoculation of contaminated blood or tissue.
This re-emergence of VBDs in humans and animals, has
presented a 'One Health' issue highlighting the
requirement for interaction between veterinary and
human medical sectors for the benefit of animal and
human health and the global environment (Braks et al.,
2007; Day, 2011; Gubler, 2009). There are many
challenges in achieving the above goals: (1) promoting
detection of these diseases in animals and humans, (2)
undertaking comparative and translational research (3)
developing robust diagnostic tests and surveillance
systems (4) studying the connection between pet animals
and wildlife reservoirs (6) promoting awareness of pet
owners on the importance of regular ectoparasite control,
and (7) developing strategies to minimize the risk
involved in pet animal mobility with regard to zoonotic
diseases. This paper intends to highlight the complex
clinical presentations associated with tick-borne diseases
(TBD) based on scientific information and personal
experiences on such diseases in dogs.
The organisms responsible for TBDs in South-East Asia
and the Indian Subcontinent are: Anaplasma
(phagocytophilum and platys), Babesia (vogeli, gibsoni
and canis), Ehrlichia (canis, chaffeensis), Hepatozooan
canis, and Rickettsia (felis, japonica, conorii, typhi &
orientia tsutsugamushi) (CVBD).
Vector-borne
hemoparasites in dogs in Sri Lanka are Anaplasma platys,
Babesia gibsoni, Babesia canis, Ehrlichia canis and
Hepatozooa canis. Presence of Rickettsial parasites (R.
conorii, R. typhi, Orientia tsutsugamushi) also had been
detected serologically (Nanayakkara et al., 2013).
Furthermore, intermittent, persistent infections or coinfections caused by Babesia, Anaplasma, Ehrlichia,
Hepatozoon, Rickettsia and Mycoplasma haemocanis
(Haemobartonella) are commonly encountered by
practitioners. Co-infections of Ehrlichia, Babesia and
Hepatozoon can occur particularly in endemic areas since
the same tick species can transmit several pathogens. Such
infections probably explain the variations in clinical
presentation, pathogenicity and response to therapy
(Shaw et al., 2001).
Babesia organisms are broadly divided by their size in
to small and large. The large babesia which were
previously known as Babesia canis are reclassified as B.
canis, B. vogeli, B. rossi and Babesia sp (Coco). The small
Babesia previously known as Babesia gibsoni are
reclassified as B. gibsoni and B. conradae; Babesia
microti as Theilaria annae; and Babesia equi as Theileria
equi (Irwin, 2009). Babesiosis in cats is less common and
is manifested as an afebrile, chronic, low-grade disease,
with anore xia, letha rgy, anemi a, depre ssion , and
occasionally icterus. Feline Babesiosis is caused by
Babesia felis, Babesia cati (India), B. canis (Europe), B.
canis presentii (Israel), B. vogeli (Thailand).
Ehrlichia and Anaplasma spp. are closely related with
four distinct clades: Anaplasma, Ehrlichia, Wolbachia and
Neorickettsia genegroups (Dumler et al., 2001; Ferla et al.,
2013).
Ehrlichia gene group- Ehrlicha canis, E. chaffeensis
and E. Ewingii
Ana pla sma g ene g rou ps- E. Phagocytophilum
(formerly known as E. equi) and E. platys (synonymn
Anaplasma platys) Ehrlichia Phagocytophilum
(Anaplasma) infection in neutrophils has become
increasingly significant in human, canine and feline
populations causing granulocytic ehrlichiosis.
Neoric kettsia gene groups - E. sennetu, E. risticii and
Neorickettsia helminthoeca
Ehrlichia chaffeensis cause human monocytic
ehrlichiosis, Ehrlichia ewingii causes human granulocytic
ehrlichiosis, and Anaplasma phagocytophilum cause
human anaplasmosis (formerly known as human
granulocytic ehrlichiosis, or HGE).
Transmission
The transmission of TBD pathogens can occur in
different ways. In biological transmission, the pathogen
http://doi.org/10.4038/slvj.v63i2.9
2
Indira Silva
undergoes some biological development in the body of the
arthropod vector in order to complete its life cycle.
Mechanical transmission is a simple transfer of the
organism on contaminated mouth parts or other body parts,
without multiplication or developmental change of the
pathogen in the arthropod (Gubler, 2009). In addition to
biological and mechanical transmission, direct
transmission of TBD can occur through dog bites,
contaminated equipment, transplacentally and via blood
transfusions. Although Ehrlichia canis and Hepatozoon
canis are transmitted by ticks, E. canis is transmitted
through the saliva of ticks during a blood meal, while H.
canis is transmitted by ingestion of a tick containing
mature sporozoites (Baneth et al., 2007; Harrus and Waner,
2011).
Hemotrophic mycoplasmas (hemoplasmas), formerly
classified as Haemobartonella and Eperythrozoon species,
with a single circular chromosome are transmitted by
Rhipicephalus sanguineus and Dermacentor reticulatus
ticks, from a blood transfusion and also directly through
saliva during a fight (Willi et al., 2006; Nascimento et al.,
2012). They adhere to the surface of the erythrocytes and
cause extravascular hemolysis. Mycoplasma hemofelis, M.
hemocanis and Candidatus are closely related (Sykes and
Tasker 2016; Braga et al., 2012). Hemoplasmas in dogs
are M. haemocanis, Candidatus M. haematoparvum
(Nascimento et al., 2012). Mycoplasma haemofelis,
Candidatus Mycoplasma haemominutum, Candidatus
Mycoplasma turicensis cause feline infectious anemia in
the presence of retrovirus-induced immunosuppression or
interaction with blood progenitor cells.
Phagolysosome
Extrinsic incubation period (time required to develop
inside the arthropod and to progress to the infective stage)
is generally 7 to l4 days (Gubler, 2009). Mitochondria are
important for the division of Babesia by binary fission (10
h in vitro) which depends on nucleic acid synthesis and
energy metabolism via aerobic and anaerobic glycolysis,
converting glucose to lactate (Rajapakshage et al., 2012).
The mitochondrial DNA (mtDNA) consists of three
functional genes essential for energy metabolism.
cytochrome c oxidase subunit I (COXI) - electron
transfer in respiratory chain
cytochrome c oxidase subunit III (COXIII) assembly and stabilization of the entire cytochrome
complex
cytochrome b (CYTb) - subunit of COXIII - electron
transfer
Ehrlichia organisms survive and multiply in the
infected cell by their ability to inhibit the fusion of the
phagosomelysosome (Park and Rikihisa, 1991; Wells and
Rikihisa, 1988).
Immunity
The cell-mediated immunity (CMI), the primary but not
exclusive, is critical for the outcome of an infection
(Waner et al., 2001). Macrophages play a leading role in
CMI by phagocytosing and killing of organisms by O2-
dependent and O2-independent mechanisms, and in
initiating an acute phase response by releasing cytokine
leading to a humoral immune response.
Cytokines contribute to both host pathology and host
immunity (Hemmer et al., 2000). The T-cell-induced
immunity and proinflammatory cytokine, Interferone
gamma (IFNa) secretion are predominant mechanisms
for recovery from and immunity to ehrlichial infections
(Waner et al., 2001). The cytokine IFNg released from
Th1 cells, macrophages and natural killer (NK) cells,
enhance Tumor necrosis factor alpha (TNFa) expression
which up-regulate the expression of adhesion molecules
in endothelia, promoting sequestration of organisms to
endothelia (Hemmer et al., 2000; Irwin, 2009; Tajima and
Rikihisa, 2005). Furthermore, the recruit of leukocytes by
TNFa lead to changes in vascular epithelium resulting in
increased vascular permeability, oedema and tissue injury.
Therefore, TNFa production is detrimental to the host as
it enhances inflammation, leading to abnormal
perfusion,tissue hypoxia, sepsis and systemic
inflammatory response (SIRS). Increased expression of
cytokines, IL-10 and IL-4 is important for resolution of
parasitemia (Hemmer et al., 2000).
The immunologic responses among monocytophilic
Ehrlichiae of the family Anaplasmataceae (E. canis,
E.chaffeensis, A. platys and A. phagocytophilum) are
similar (Brouqui and Dumler, 2002). Polyclonal
hypergammaglobulinemia comprising IgM and IgA
appear in 2-7 days, and shift to IgG by two weeks postinfection (Iqbal et al., 1994; Weisiger et al., 1975). A titre
of =1:160 in 2-4 weeks can rise to 1:2560 - 1:20,480 in >4
weeks. Declining titres may indicate recovery while very
high gammaglobulinemia can pose a risk of
hyperviscosity causing nervous signs. The CD4:CD8 ratio
is a deciding factor for pathogen elimination or survival.
The cytokines from Th1 cells (IFNg & TNFa) result in
elevation in IgG 2 which is responsible for
pathogenelimination (Nair et al., 2016). An infection
causes immunodysregulation reducing lymphocyte
blastogenesis and decreasing CD4:CD8 ratio yielding low
Th1-cytokines which will result in elevation in IgG1 which
is poor in pathogen specific, and therefore cause severe
infection. Quantitative ELISA has shown a predominant
elevation in IgG2 in E. canis infection when compared
with E. chaffeensis, A. platys and A. phagocytophilum
infections. A dog with clinical signs and hematological
parameters suggestive of Chronic Monocytic Ehrlichiosis
(CME) with a single serum IgG IFA titer of 1:256 can be
defined as a “definitive” case, while a “probable” case
would be one with clinical signs and symptoms and a
single IgG IFA titer of 1:64-1:128 (Waner et al., 2001).
However, hypogammaglobulinemia has been seen in
pancytopenic dogs (Waner et al., 2001). Marked
pancytopenia due to bone marrow hypoplasia is a
hallmark of chronic severe Ehrlichiosis.
Cross-reactions between apicomplexan parasites and
operator subjectivity may limit the benefits of serology.
Cross reactivity between ehrlichiosis, anaplasmosis, or
Tick-borne diseases
neorickettsiosis infections can be distinguished by
western immunoblotting (Harrus and Waner, 2011). There
is no serological cross reactivity between E. canis and
Anaplasma platys, and little cross reactivity, if any, with
Rickettsia rickettsii, of Rocky Mountain Spotted Fever
(Harrus and Waner, 2011).
Persistent titres may occur in persistent infections,
reinfection, treatment failure, and an abnormal immune
response. It has been speculated that dogs infected with
vector-transmitted pathogens may develop persistent
infection within systemic circulation and therefore may be
more prone to develop autoi mmune anti-nuclear
antibodies (ANA) (Smith et al., 2004). It has been
suggested that in persistent VBD infections, prolonged
antigenic stimulation due to ineffective apoptosis or
defective removal of cell debris by neutrophils and
macrophages might lead to production of perinuclear
antineutrophil cytoplasmic autoantibodies (pANCA)
(Karagianni et al., 2012).
Detection of parasites in blood and tissues
Detection of piroplasms can be done morphologically,
serologically or using molecular techniques. Cell culture
isolation is expensive and time consuming. Serology may
not differentiate acute from chronic phases, and testing
need to be done in experienced diagnostic labs with
str ing ent qua lit y con tro l mea sur es. Cyt olo gic al
examination of thin blood films by light microscopy lacks
both sensitivity and specificity and depends on the degree
of parasitaemia. It might be difficult to detect Babesia
through light microscopy when hemolytic anaemia or
thrombocytopenia is present. However, the probability of
detection of Babesia is high in smears made from capillary
beds (ear tip, toe nail) or from cells beneath the buffy coat.
Detection in carrier dogs can be unrewarding due to very
low, often intermittent parasitaemias (Irwin, 2009).
The morulae of Ehrlichia and anaplasma can be
detected either in mononuclear cells, neutrophils or
platelets, depending on the organism. The detection of
morulae could be optimized by examining buffy coat
smears (1000 oil immersion fields)(Harrus and Waner,
2011). Precautions must be taken not to confuse ehrlichial
inclusions with platelets, lymphocytic azurophilic
granules, and phagocytosed nuclear material when
examining blood smears. Cases may be erroneously
diagnosed as lymphocytic leukemia as a large proportion
of lymphocytes can show prominent azurophilic granules,
cleaved or indented cell nuclei with mature nuclear
chromatin, consistent with large granular lymphocytes
(Heeb et al., 2003).
Although the detection limit of light microscopy is
approximately 0.001% parasitaemia for Babesia (Böse et
al., 1995) and only 4% of blood smears for E. canis
(Woody and Hoskins, 1991), Polymerase Chain Reaction
(PCR) has a high sensitivity and specificity in parasite
detection when there are approximately 50 organisms/ml
(Birkenheuer et al., 2003) and 9 parasites/ ìl (Matsuu et al.,
2005). However, fluctuating parasitemia can limit
parasite detection by PCR testing. A one-step multiplex
3
PCR has been developed for simultaneous detection of
Ehrlichia canis, Babesia spp and Hepatozoon canis, from
blood samples in a single reaction (Kledmanee et al.,
2009). The primers used are specific to E. canis VirB9,
Babesia spp 16S rRNA and H. canis 16S rRNA genes. The
sensitivity in detecting H. canis in buffy coat, blood and
bone marrow is high with PCR compared with cytology
using the same tissue (Otranto et al., 2011).
General Clinical findings
Critical determinants of infection severity are virulence
of the organism, age, gender and the immune status of the
patient. The records of the Veterinary Teaching Hospital
(VTH) reveal that 34% of the dogs with TBD were less
than one year old, and 52% of them were less than 6
months old. Males were at higher risk of infection with
M:F ratio of 1.5. Approximately 28% of males was
critically anemic (HCT <10%), compared with 14%
females.
Rottweilers, Labradors, Boxers and cross bred dogs of
any age were more susceptible for Babesiosis and
Ehrlichiosis than other breeds. However, mostly young
ad ul ts of Ge rm an sh ep he rd s, Do be rm an n, an d
Pomeranian s showed higher susceptibi lity. It was
assumed that the cross breds were at a higher risk probably
because the probability of getting infected is high as they
were mostly free ranging and also more exposed to dog
bites during fights.
The degree of parasitemia depends on the immune
status of susceptible dogs. When infected with the KR-1
strain of B. microti, the SCID (Severe Combined
Immunodeficiency) mice and TCR knockouts (T cell
receptor-beta-deficient mice) had shown to sustain
severe parasitemia; IFN-ã deficient mice had
developed a less severe parasitemia (Clawson et al.,
2002). In contrast, the levels of parasitemia in JHD-null
mice which lack B-lymphocytes and antibodies were
indistinguishable from the wild-type animals (Clawson et
al., 2002). These data indicate that cellular immunity is
critical for the clearance of B. microti.
Clinical signs and hematological changes, such as,
hyperthermia, severe/critical anemia, thrombocytopenia,
hypoalbuminemia, lymphadenopathy, hepatomegaly,
splenomegaly and evidence of internal bleeding are not
specific to a single tick-borne pathogen. The overall
clinical and haematological findings propose a multisystem disease complex, and therefore, the findings
should be carefully interpreted to avoid incorrect
diagnosis and inappropriate treatment. Common clinical
signs seen in TBDs are:
• Hyperthermia 104-106 °F (40-41°C), Sometimes
hypothermia in Babesiosis (98.7°F)
• Anemia severe/critical, Evan's syndrome (IMHA/
IMTP)
• Leukocytosis/ Leucopenia
• Thrombocytopenia with or without clinical bleeding
such as, epistaxis, hematuria
• Hypoalbuminemia
4
Indira Silva
• Tachycardia (up to 80/min), elevated CRT
• Tachypnoea (up to 80/min), dyspnoea, pulmonary
oedema
• Lymphadenopathy, splenomegaly, hepatomegaly,
liver enzyme, jaundice, bilirubinuria
Occasional signs seen in Ehrlichiosis are;
• Neurologic (ataxia, seizures, paraparesis/ tetraparesis
[upper/lower motor-neuron deficits], stupor,
vestibular disease)
• Polyarthritis (stiffness, swollen joints, reluctance to
move [E. ewingii and A. Phagocytophilum])
• Ocular lesions (chorioretinitis, retinal detachment,
uveitis)
• Vomiting, diarrhea
Systemic Findings
The vector-borne parasites cause severe damage to
various organs in the host. The highest pathological
grading have been seen in E. canis infected dogs followed
by E. chaffeensis, A. platys and A. phagocytophilum,
respectively (Nair et al., 2016). Chronic ehrlichiosis is
associated with irreversible bone marrow destruction
(Skotarczak, 2003). The infection status of E. canis and E.
chaffeensis can be detected better with PCR testing of
spleen and lymphnodes compared to A. platys and A.
phagocytophilum (Nair et al., 2016). Detection of
Ehrlichia DNA in tissues may not necessarily correlate
with the presence of viable organisms, as Ehrlichial DNA
can be found in blood or haemolymphatic tissues in dogs
in which the infections are at sub-clinical level or has
contributed to other disease conditions and masked by the
presenting condition (Gal et al., 2008). It has been shown
experimentally that Ehrlichial DNA could not be detected
from blood or spleen after 9 and 60 days of treatment with
doxycycline, respectively (Harrus et al., 2004). Therefore,
it can be presumed that DNA from dead E. canis
organisms gets probably cleared up in dogs and does not
persist in the tissues for a long time after the elimination of
infection by successful treatment or by an effective host
immune response.
Anemia and Fever
Anemia caused by hemolysis, internal hemorrhage, or
poor erythropoiesis could be critical leading to hypoxemia.
The hemolysis could be intravascular and/or
extravascular and either immune-mediated (IMHA) or
non-immune mediated. Both intra and extravascular
hemolysis lead to fever, icterus with elevated bilirubin.
Ghost cells, autoagglutination and hemoglobinemia (cellfree plasma hemoglobin indicated by an elevated mean
corpuscular hemolysis, MCHC), and hemoglobinuria are
evidences of immune-mediated intravascular hemolysis.
Erythrophagocytosis and spherocytes, indicating
complete and partial phagocytosis, respectively, are
evidence of immune-mediated extravascular hemolysis
where hemolysis occurs in the mononuclear phagocytic
system (MPS). Hemolysis due to non-immune mediation
occurs as a result of oxidative damage, intensive lipid
peroxidation due to generation of reactive oxygen species
(ROS) molecules in parasitized erythrocytes, increased
membr ane perme abili ty and decre ased membr ane
potential. Malondialdehyde (MDA), an end product of
ROS excreted in the urine, blood, and body fluids
indicates oxidative stress and non-immune mediated
hemolysis in B. gibsoni, B. canis and in experimental
mixed infection of Ehrlichia canis and B. gibsoni
(DVM360.com). Elevated MDA is a prognostic marker of
disease severity and outcome.
In most occasions, a marked drop in HCT could be seen
before parasitemia is detected in peripheral blood. This
happens when erythrocyte destruction is greater than the
degree of parasitemia indicating lysis of both parasitized
and non-parasitized erythrocytes. The compensatory
response to anemia by the bone marrow is evident by
polychromasia indicating the presence of reticulocytes.
The bone marrow response is classified as nonregenerative, or decompensatory, when nucleated
erythrocytes (NRBC) are evident in peripheral blood,
indicating the need to elevate the RBC volume by
stimulating erythropoiesis or transfusing erythrocytes in
order to alleviate hypoxemia. However, blood
transfusions may not be possible in IMHA due to the
presence of anti-erythrocytic antibodies which would lyse
the transfused cells. Of the rickettsial organisms, E. canis
cause the most significant drop in the HCT when
compared with E. chaffeensis, A. phagocytophilum and A.
platys (Nair et al., 2016). Samples with autoagglutination
may record an elevated red cell distribution width (RDW).
The immune driven hypoproliferative anemia (Anemia
of Inflammatory Disease, AID) could be another finding
in Babesiosis and Ehrlichiosis. In AID, the availability of
iron get reduced as a result of cytokine activation which
inhibit duodenal Fe absorption, increase Fe uptake and
retention in macrophages, reducing erythropoietin
production in the kidneys, and directly inhibiting
erythroid progenitor cells (Weiss and Goodnough, 2005).
Bone marrow erythropoeisis can get impaired due to
cytokine-mediated apoptosis, down-regulation of
erythropoeitin receptor expression on erythroid
progenitor cells, poor expression of other prohaematopoietic factors, and low Fe availability for
hemoglobin synthesis (Weiss and Goodnough, 2005) .
Hypoferremia despite adequate Fe stores is a host
response to limit microbial access to Fe. This inhibition of
pathogens due to limited access of iron, has been
demonstrated in Ehrlichia (Park and Rikihisa, 1991).
When comparing the pathogens of the family
Anaplasmataceae, pronounced anemia and reduction in
PCV was seen in E. canis infections, and severe
thrombocytopenia was manifested with the platelettrophic A. platys (Nair et al., 2016).
V a s c u l o p a t h y, H e m o c o n c e n t r a t i o n a n d
Hypercoagulability
Following intravascular hemolysis, the
decompartmentalized hemoglobin or cell-free plasma
oxyhemoglobin causes irreversible oxidation of
endothelial NO resulting in vasomotor instability and
Tick-borne diseases
vasculopathy (Minneci et al., 2005; Palmer et al., 1988;
Rother et al., 2005). The NO regulates homeostatic
vascular functions such as vasodilation, inhibition of
plat elet acti vati on and thro mbos is, inhi biti on of
endothelial adhesion molecule and endothelin expression,
and modulation of intimal and smooth muscle
proliferation (Furchgott and Zawadzki 1980).
High mortality may occur as a result of loss of
intravascular fluid due to increased capillary permeability,
causing the extravascular protein-rich fluid compartment
to raise leading to hemoconcentration and hyperviscosity,
known as “red biliary”. This state will be clinically
evident either as an absolute elevation of HCT from severe
or critical anemia to over 30% or a normal HCT that is
inappropriate for the degree of hemolysis (relative
elevation of HCT) (Pardini, 2000).
Haemoglobin-mediated oxidative damage of
endothelia during intravascular haemolysis is generally
prevented by Hemopexin in plasma (Kuleš et al., 2014).
Endothelial damage can also occur by reactive oxygen
intermediates (ROS), activation of the complement
cascade and cytokine activation on endothelia, as a result
of autoagglutination, circulatory stasis and sequestration
of parasitized cells. Endothelial phospholipase damage
activates the coagulation cascade leading to the
hypercoagulable state of disseminated intravascular
coagulation (DIC) and formation of micro-thrombi.
Thromboelastography (TEG) and thromboelastometry
(ROTEM) can detect hypercoagulable states (Wiinberg et
al., 2005). The TEG gives a graphical representation of
clot formation and lysis, evaluating precoagulation,
coagulation, and fibrinolysis.
The micro thrombi formed from DIC can lead to venous
thrombosis, obstruction of vessels and tissue ischemia,
particularly in the brain and lungs, thus producing clinical
signs associated with pulmonary thromboembolism
(PTE) and nervous system. The coagulation panel will
reveal thromboc ytopenia and hyperfib rinogene mia,
increased circulating D-dimers, shortened PT or aPTT
(Song et al., 2016). The consumption coagulopathy
resulting from DIC will exacerbate hemorrhage caused by
direct vasculopathy and thrombocytopenia, thereby
increasing the risk of mortality due to hemorrhages.
Plasma may reveal hypofibrinogenemia, elevated APTT,
PT, FDP, D-dimer, and buccal mucosal bleeding time. The
hemorrhaging will be clinically evident as epistaxis,
hematuria, petechia, and melena as single or multiple
manifestations. The survival for dogs with aortic
thrombosis (ATh) or aortic thromboembolism (ATE)
would be between 50% and 60%, and those with chronic
clinical signs have a better prognosis than those acutely or
severely affected (Williams et al., 2016). The prevalence
of PTE in dogs is underestimated in necropsies, since
thrombi in dogs lyse more rapidly than in humans (within
3 hours of death) due to greater plasminogen activator
activity, greater platelet lytic activity, and secretion of
plasminogen activator by the pulmonary endothelium
(Goggs et al., 2009).
5
Thrombocytopenia
Severe thrombocytopenia is a consistent finding in the
acute phase of Babesiosis. Thrombocytopenia, either
constant or cyclic, is an almost consistent finding in
Ric ket tsi al inf ect ion s, but it can not dis tin gui sh
Ehrlichial/anaplasma infections (Nair et al., 2016).
Thrombocytopenia can be critical or severe, with or
without clinical bleeding. Significant persistent
thrombocytopenia is observed with platelet-trophic A.
platys, and also E. canis and A. phagocytophilum
infections even though cell tropisms for the latter two
pathogens are monocytes and granulocytes, respectively
(Nair et al., 2016) . Thrombocytopenia from A. platys
infection is a result of pathogen antigens, whereas
thrombocytopenia in Ehrlichiosis is due to anti-platelet
antibodies, but not due to anti-ehrlichial antibodies (Nair
et al., 2016), because E. canis alters the immune system to
overproduce natural anti-platelet antibodies with high
affinity (Harrus et al., 1998; Harrus et al., 1996).
Decreasing platelet counts correlate well with increasing
anti-thrombocyte IFA titres and PCR positivity for E.
canis DNA 16S rRNA, highlighting the importance of
evaluating the platelet counts in suspected dogs (Harrus
and Waner, 2011; Harrus et al., 1998; Harrus et al., 1996;
Köster et al., 2015). It is important to rule out platelet
agglutination in blood smears when evaluating the actual
platelet count. Immune mediated thrombocytopenia
(IMTP) occurring together with immune mediated
hemolytic anemia (IMHA) is identified as the Evans
Syndrome.
Evidence of bleeding without severe thrombocytopenia
could be due to thrombocytopathia causing reduced
platelet aggregation, reduced platelet adhesiveness,
Interference in PF3 release and platelet migration
inhibition causing reduced pseudopod formation.
Hypoxemia and Hypoperfusion
The pathophysiological sequelae induced by PTE result
in hypoxemia, hyperventilation, and dyspnea. The arterial
hypoxemia secondary to multiple abnormal ventilation:
perfusion (V:Q) ratios in the affected lungs, can get
complic ated fro m inters titial a nd alveo lar edem a
(congestive atelectasis) causing diffusion impairment,
airway closure and alveolar collapse. An occlusion of
>60% of the pulmonary vasculature will increase
pulmonary vascular resistance (PVR) which will reduce
the pulmonary arterial flow(Ebert et al., 1967). Reflex
vasoconstriction secondary to alveolar hypoxia may also
contribute to elevated PVR leading to pulmonary arterial
hypertension (PAH) and increased right ventricular (RV)
afterload, RV dilatation and dysfunction (Goggs et al.,
2009). The reduction in left ventricular filling as a
consequence of RV dilatation will decrease cardiac output
leading to signs of forward failure (hypotension,
cardiogenic shock). If the patient survives an acute crisis
the residual pulmonary hypertension will lead to a long
term backward failure resulting in hepatomegaly, ascites,
and pleural effusion.
Indira Silva
6
The increased vascular permeability due to
vasculopathy will cause tissue oedema and O2 extraction
deficit leading to severe hypoxemia and hypovolemia.
These mechanisms finally lead to severe hypoperfusion,
clinically evident as elevated capillary refilling time
(CRT). The resulting hypoperfusion will demand a change
in the distribution of the cardiac output (Q) in order to
maintain perfusion to vital organs and tissues, namely
heart, lungs, brain, diaphragm, and intercostal muscles
(Adachi et al., 1976). Such changes in the presence of
restricted coronary blood flow and coronary sinus PO2
could be achieved by increasing myocardial O2
consumption and efficiency, and the cardiac work. The
hemodynamic and metabolic changes in such patients
would be bradycardia, hypotension, reduced cardiac
index [CI = LV Q/ BSA (mL/min/m2)], reduced shock
index (i.Shock =HR/SBP), mild changes in pulmonary
capillary pressure (PCP) and central venous pressure
(CVP), reduced mixed venous saturation (SvO2, percent
O2 bound to HB in right atrium), reduced venous O2
pressure (PvO2), reduced O2 transport, while increasing
O2 consumption (VO2), O2 extraction (TEO2), and serum
lactate. The prognosis of such patients would be grave due
to generalized edema and pulmonary edema (greater
mortality), ultimately leading to cardiac failure and
multiple organ dysfunction. Such patients may show
evidence of hepatic dysfunction (hypoalbuminemia,
markedly elevated ALT, elevated bilirubin, icterus and
bilirubinuria), impaired gas exchange & respiratory
dysfunction (dyspnea, tachypnoea up to 80/m),
pulmonary edema, acute respiratory distress syndrome
(ARDS), and renal azotemia which will drastically
increase the risk of mortality (Harison et al., 2012). The
systemic vasoconstriction resulting from vasculopathy
and hypovolemia will further impair renal function
leading to renal failure.
Clinical Biomarkers of tick-borne diseases
Clinical biomarkers signify multisystemic pathologies
(Diniz et al., 2008; Köster et al., 2015; Koutinas et al.,
2012).
Blood Profile- IMHA. Hemolytic anemia,
Thrombocytopenia, Hemoconcentration
Coagulation profile- Thrombocytopenia,
consumption coagulopathy (­APTT, PT, D-dimer)
Hemochemical abnormalities - metabolic acidosis
& respiratory alkalosis, Hyperlactatemia,
Hyperbilirubinemia of >170 ìmol/L, elevated
ALT
Cardiac- Elevated cardiac troponin I (cTnI)
concentrations. The ECG abnormalities were not
associated with disease severity, outcome, or plasma
cTnI concentrations, other than the ventricular
premature complexes associated with high cTnI.
Respiratory -Pulmonary edema.
For acute and chronic diseases of Ehrlichia and
anaplasma, the carriers can be categorized according to
results of multiple tests, as shown in Table below. Crossreactions are common in serological tests such as IFA,
dot-ELISA (“Immunocomb”). Nested PCR (nPCR) by
16S rRNA amplification, detect DNA earlier than
serology, and cross-reactions are uncommon.A p30-based
nested PCR assay has been developed for detection of E.
canis in both dog and ticks(Stich et al., 2002). The
coombs' test cannot diagnose IMHA because it lacks
sensitivity.
Most dogs recover from clinical signs with treatment
but no single drug had been successful in completely
eliminating the organisms, and thus requiring adjunctive
treatment, including O2 therapy, immunosuppressants,
antithrombotics, antimicrobials and fluid. Antibabesiacidal drugs act by many different mechanisms.
Atovaquone (ubiquinone analog) block mitochondrial
electron transfer leading to reduced energy production for
parasites. Berenil (Diminazene aceturate) interferes in
aerobic glycolysis (reduce energy production) and
multiplication (interact with DNA minor groove). The
Berenil resistant genes (CYTb) can transfer during
pathogen multiplication, and the proliferation potential
and the degree of parasitemia is low in Berenil resistant
Babesia strains. Berenil toxicity can occur from
overdosing, repeated dosing, and even with
recommended doses on Babesia negative animals. The
anticholinergic anthelmintic, Imidocarb (carbanilide) can
be used in most tick-borne diseases. Drug combinations
have shown to be either effective or suppress parasitemia
below the limit of detection, thereby reducing the
transmission of protozoa through vectors.
Ehrlichia organisms survive and multiply in the
infected phagocyte by their ability to inhibit the fusion of
the phagosomelysosome (Park and Rikihisa, 1991; Wells
and Rikihisa, 1988). Treatment of Ehrlichiosis with
doxycycline, an antibiotic which inhibits prokaryote
protein synthesis, restores phagosomelysosome fusion
(Wells and Rikihisa, 1988).
Table 1. Interpretation of laboratory findings in Ehrlichiosis
Serology
nPCR
WBC
Interpretation
Positive
Negative
Positive
Positive
Positive
Positive
Negative
Negative
Leukocytosis
Leukocytosis
Pancytopenic
Acute disease (Morulae)
Acute disease
Chronic disease
Persistent titres (Carriers/ Treated)
Tick-borne diseases
ACKNOWLEDGEMENTS
The author acknowledges the clinical and laboratory
contributions of Dr. WCR Fernando, Ms. MRCK Mallawa
of the Veterinary Teaching Hospital.
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S.L.Vet.J. 2016, 63:2(A): 11-16
Original article
ASSESSMENT AND COMPARISON OF URINARY PROTEIN:
CREATININE RATIOS OF HEALTHY DOMESTIC DOGS AND
WORKING DOGS IN KENNELS DIVISION IN SRI LANKA POLICE FOR
EARLY DETECTION OF RENAL DISEASE
1
2
3
K.A.N. Wijayawardhane , K.G.S.S. Karunathilake and V.S.S Kumara
1
Department of Veterinary Clinical Sciences, Faculty of Veterinary Medicine and Animal Science,
University of Peradeniya
2
Beloved Animal Hospital, 264/4, Miriswatta
3
Provincial Director's Office, Department of Animal Production and Health, Mannar
Summary: Healthy dogs usually excrete small amounts of protein in urine. Persistently high proteinuria is
usually a marker of kidney disease. The urinary protein: creatinine ratio (UPC) is used in quantifying
urine protein excretion in order to diagnose kidney diseases, guiding recommendations for monitoring and
treatment of kidney diseases and evaluation of prognosis. However, use of UPC ratio has not been well
established/ used to diagnose renal diseases in dogs in Sri Lanka. Therefore, for the first time in Sri Lanka,
we used this method to detect UPC ratio of healthy dogs (n=51). Once we established the UPC ratios of
healthy dogs, UPC ratios were measured in working dogs (n=45) in Kennels Division of Sri Lanka Police,
since renal diseases are a common cause of mortality in these dogs. In control group, the mean UPC ratio
was 0.06±0.05. This was similar to the UPC ratio of tracking group among working dogs. Two out of 22
dogs used for explosive duties had UPC ratio within the borderline proteinuric stage (0.2-0.5) according to
the staging system implemented by the International Renal Interest Society (IRIS). In the narcotic group 2
out of 12 dogs were borderline proteinuric and one dog was proteinuric. In this study, we have successfully
established the measurement of UPC ratio to detect proteinuria in Sri Lankan dogs and have identified
dogs that are susceptible to renal diseases.
INTRODUCTION
Proteinuria is one of the risk factors of renal diseases. In
addition to this, proteinuria is used as a predictor of end
organ damage (Barnas et al., 1997). Early and accurate
detection of persistent renal proteinuria is of high
importance to reduce mortality in both dogs and cats due
to renal damage (Littman, 2011; Wehner et al., 2008). By
detecting acute renal disease early in its course,
appropriate intervention can be made to arrest or at least to
attenuate renal damage and the development of acute renal
failure. Similarly, by detecting chronic renal disease
before the onset of renal azotemia and chronic renal
failure, appropriate intervention can be made to stabilize
renal function or to slow its progression (Jacob et al.,
2005). Once a complete diagnosis is made, therapeutic
and management plans can be arranged to manage the
disease. The urine protein creatinine (UPC) is the gold
standard tests to quantify and monitor proteinuria in dogs.
UPC ratio detects persistent proteinuria and aid in making
clinical decisions and monitoring response to therapy.
UPC values recommended by American College of
Veterinary Internal Medicine (ACVIM) (Lees et al., 2005)
and International Renal Interest Society (IRIS) for
azotemic and non azotemic dogs provide guidance in
monitoring, diagnosing, and treating dogs and cats with
renal disease (Brown et al., 2013; http://www.iris-
kidney.com). We have observed higher mortality rates in
working dogs in Police and armed forces in Sri Lanka, due
to renal diseases. Therefore, the primary goal of this study
was to establish the reference for UPC ratio for dogs in Sri
Lanka, and to compare UPC ratios of working dogs in Sri
Lanka Police to detect the dogs with possible renal
diseases early.
MATERIALS & METHODS
Design and study population
Fifty one client-owned, apparently healthy dogs
presented to the Veterinary Teaching Hospital (VTH),
University of Peradeniya, identified as reference group,
were used to establish the UPC ratio. These dogs were
included in the study only after signed consent had been
received. Their average age was 2.8 years (range 1-10
years). All the working dogs (n=45) except three puppies
(< 6 months) and six dogs attached to Presidential Security
Division of Sri Lanka Police were used in this study. Their
average age was 4.3 years (range 2-8 years). This group
comprised of dogs assigned for explosive detection,
narcotic detection and tracking duties. Dogs were enrolled
for this study using the following inclusion criteria: (1)
more than six months of age, any breed, gender, size and
(2) were either healthy animals, or working dogs with or
without underlying disease/s that could induce proteinuria.
http://doi.org/10.4038/slvj.v63i2.10
12
Wijayawardhane et al
A single random mid-stream sample of 10 ml urine
(Monroe et al., 1989) was collected aseptically into sterile
tubes from each animal by catheterization and transported
in ice immediately after collection. Urine samples from
animals with active sediment were excluded from the
study. An inactive urine sediment for this study was
defined as <5 WBC/ hpf, <20 RBC/ hpf, and no visible
bacteria (Duffy et al., 2015). Humantrol quality control
sera (Human GmbH-Max-Plank-Ring, Germany) were
used to check the experimental procedures.
Blood samples
Venous blood sample (2.5 ml each) was collected into
EDTA tubes from cephalic vein. Plasma was separated
within 30 minutes from collection. Hematologic and
plasma analytes were determined within 12 hours with
standard methods. Samples were analyzed for Full Blood
Count, Blood Urea Nitrogen (BUN), plasma creatinine,
total plasma protein and albumin according to the
standard diagnostic laboratory procedures established at
the VTH. Blood smears were prepared and checked to
exclude tick-borne parasitic infections.
Urinalysis and UPC measurement
Urine samples were used to estimate specific gravity,
protein and creatinine concentrations, and subjected to
microscopic sediment evaluations, within 12 hours of
collection (Rosi et al., 2012). Samples were assessed
visually for evidence of debris, or gross discoloration
(Vaden et al., 2004). Urine specific gravity was measured
using a refractometer. Multistix Uripath strips were used
according to the manufacturer's guidelines to detect
glucose, bilirubin, ketones, specific gravity, occult blood,
pH, protein, nitrite and leucocytes in urine by assessing
the colour change at recommended times after immersing
the dipstick in the sample of urine. Urine sediment, 0.5 ml,
harvested by centrifuging 5 ml of urine sample at 2000
rpm for 5 minute was evaluated microscopically at low
power (10x) for epithelial cells, casts and at high power
(40x) for leukocytes, erythrocytes, bacteria and crystals.
Urine protein concentration
Total protein concentration (mg/dl) in urine was
measured in the supernatant obtained after centrifugation
by Pyrogallol red- molybdate method (Johnson et al.,
1999) on a semi-automated biochemistry analyzer (Erba
Mannheim, ERBA Diagnostics, North Miami Avenue
Miami, U.S.A). In this method pyrogallol red/molybdate
formed a red complex in the presence of proteins and the
colour was directly proportional to the protein
concentration.
Urine creatinine concentration
Urine creatinine concentrations (mg/dl) were measured
using RANDOX Monza reagent with the semi automated
chemical analyzer using a modified Jaffe method (Bartels,
1972). In this colorimetric method creatinine in alkaline
solutions reacts with picric acid and forms a coloured
complex which is proportional to the creatinine
concentration.
Blood Urea Nitrogen concentration
Blood urea nitrogen concentr ation (mg/dl) was
mea sur ed usi ng ure ase -Be rth elo t met hod usi ng
RANDOX reagent with the semi automated chemical
analyzer. In this colorimetric method ammonia which is
produced by urea in serum in the presence of urease is
measured photometrically.
Albumin concentrations in plasma
Albumin concentrations (mg/dl) in plasma were
measured using RANDOX Monza reagent with the same
automated chemical analyzer. In this colorimetric method
the measurement of serum albumin is based on its
quantitative binding to the indicator bromocresol green.
Classification
IRIS staging classification was used to classify UPC
ratios <0.2 as nonproteinuric, =0.2 and <0.5 as borderline
proteinuric and =0.5 as proteinuric. Clinical response
category based on the ACVIM consensus statement (Lees
et al., 2005) was also assessed with the serum creatinine
concentrations. For nonazotemic category, “no action”
was taken for dogs with UPC <0.5, monitoring for kidney
function was advised for those who had UPC =0.5 and
further investigations to assess the kidney function were
recommended for dogs with UPC =1 and <2 and medical
intervention for UPC with =2. Azotemic dogs with UPC
<0.5 was considered as “nonintervention” category and
UPC =0.5 was considered as “intervention” category for
management of the condition.
Health records of Working dogs in Kennels division of
Sri Lanka Police
In a retrospective study, carried out in parallel to this
study, we collected data of working dogs in the police for
past two years.
Statistical Analysis
Data are expressed as mean±SD, and 95% CIs were
calculated. All analyses were performed with MINITAB®
Release 14.1 software. Results obtained for control group
and working dogs were compared by using the two sample
t test and ANOVA. The Regression test was used to assess
a possible relationship between UPC ratio and other
factors. Values of p<0.05 were considered significant.
RESULTS
Population
Urine samples were obtained from 51 client-owned
dogs. Samples from all of these dogs were included in the
study, since they had inactive urine sediment. Number of
dogs in the kennels division of Sri Lanka Police had
variable degrees of active urine sediment and the samples
which were compatible with our inclusion criteria were
included in the study (Figure 1).
13
Urinary protein: creatinine ratios of dogs
Figure 1. Presence of active urine sediment among different groups of dogs in the study
population
Table 1: Blood urea nitrogen (BUN), plasma concentrations of creatinine, total proteins (TP), albumin
(Alb), urine specific gravity (USG), urine pH and urine protein creatinine ratio (UPC) of samples
collected from healthy dogs
BUN
11.46±6.44
Creatinine (mg/dl)
1.09±0.29
TP (g/dl)
5.85±1.08
Albumin (g/dl)
2.47±0.38
USG
pH
UPC
1.02±0.01
6.44±0.69
0.06±0.05
BUN- Blood urea nitrogen; TP- Total protein; USG- Urine specific gravity; UPC- urine protein: creatine ratio
Urinalysis of control group
Average UPC ratio of 51 healthy animals in the control
group was estimated to be 0.06±0.05 (Table 1).
Results obtained using health records of working dogs in
kennels division of Sri Lanka Police showed that twenty
one deaths have been reported and necropsies performed
at the Division of Pathology of the Department of
Pathobiology, and Division of the Diagnostic Pathology
of the VTH, Faculty of Veterinary Medicine and Animal
Science, University of Peradeniya. Necropsy reports
indicated that the highest number of deaths was due to
renal failure (33.3%). In addition to this, haemoparasitism
(24.7%), heart failure (19%), multi organ dysfunction
(9.5%), toxicosis (9.5%) and lymphosarcoma (4%) were
associated with the deaths of working dogs in police.
Results of UPC ratios in control group and working
dogs
Mean and standard deviation of UPC ratio in healthy
dogs was 0.06±0.05 (n=51). Mean and standard deviation
of UPC ratio in dogs in explosive (n=22), narcotic (n=12)
and tracking (n=9) duties were 0.09±0.09, 0.15±0.26 and
0.05±0.06 respectively. One way ANOVA revealed a
significant difference between mean UPC values in
control group and working dogs (p<0.05, R-Sq=9.91%).
Two sample t- tests were used to compare UPC values
between groups. Results showed that there were
significant differences (p<0.05) between UPC ratios in
dogs in the control group vs explosive and narcotic groups.
There was no significant difference (p>0.05) in UPC
value of dogs in the control group vs tracking group.
One of the key objectives of this study was to screen the
renal diseases of working dogs in the kennels division of
Sri Lanka Police. Therefore, rather than comparing the
mean UPC ratios between groups, we compared UPC
ratios of individual animals in the study population with
the IRIS standards recommended by ACVMA (Figure 2).
In the narcotic group, one animal was proteinuric but
nonazotemic. According to the ACVIM co.nsensus
statement this animal was subjected for monitoring of
kidney function. Two animals in the narcotic and two in
the explosive groups were borderline proteinuric hence no
action was taken (http://www.iris-kidney.com/) (Table 2).
Wijayawardhane et al
14
Figure 2: UPC ratios in control group and different categories of working dogs in Sri Lanka Police. In the narcotic
group, one animal was proteinuric (marked with a red asterisk). Two animals in the narcotic and two in the
explosive groups were borderline proteinuric (marked with blue asterisks)
Table 2: Matching the stage of proteinuria, blood urea nitrogen (BUN), plasma creatinine, Urine specific gravity
(USG), Urine pH, urine protein (U. protein) in working dogs that were ³ borderline proteinuric, according to
the ACVIM consensus statement
Group
BUN
P. creatinine
USG
Urine pH
U. Protein (mg/dl) UPC ratio
1.03 ± 0.01
1.02 ± 0.01
6.66 ± 1.15
7 ± 1.41
533.33 ± 404.2
100
(mg/ dl)
Narcotic (n=3)
Explosive (n=2)
17.68 ± 5.8
16.29 ± 2.59
1.31 ± 0.23
1.42 ± 0.21
DISCUSSION
An increase in urinary protein excretion is a widely
accepted tool in the detection, diagnosis, and management
of renal disease (Price et al., 2005). The UPC ratio is the
gold standard test to quantify proteinuria in dogs (Harley
and Langston, 2012). However, in Sri Lanka there had
been no evidence of using this test to stage renal diseases
in dogs. Therefore, in this study we were able to
successfully assess and establish UPC ratios in a cohort of
healthy dogs in Sri Lanka. All the animals sampled in the
control group were nonproteinuric and had no sign of
active urine sediment. The differences in UPC values
among the animals in the control group would have been
due to the time of the day of urine samples collected as one
report suggested that the first morning specimens are
preferred to detect protein excretion (Xin et al., 2004).
0.46 ± 0.41
0.35 ± 0.03
However, there are several reports supporting the idea that
use of random urine samples for the measurements of
UPC ratios work accurate and more practical (Newman et
al., 2000; Price et al., 2005).
Study of necropsy records (from 2012-2014), of
working dogs in Kennels division of Sri Lanka Police
supports the fact that renal failure is the major cause for
deaths. Bringing up a working dog to their working
capacity from the time of importation is quite an
expensive task. Therefore, detecting renal function at
early age is of high importance for successful monitoring
and therapeutic interventions. In our survey, we have
found that even young working dogs (age 2-5 years) have
died of renal disease. Chronic renal failure has been the
cause of death (~71%) in many of these animals diagnosed
with renal disease. Although, the cases of renal failure are
presented towards the latter part of their illness, proper
15
Urinary protein: creatinine ratios of dogs
screening of kidney function in dogs using UPC ratio
would significantly reduce the mortality. It is not just the
animals with kidney disease that should be screened for
proteinura. According to the author's experience, working
dogs in the Sri Lanka police are very prone to have
con dit ion s or dis eas es suc h as dru g rea cti ons ,
hyperadrenocorticism, acute renal failure, viral disease,
imm une med iat ed dis eas es, tic k-b orn e dis eas es,
exogenous steroid use, and urogenital diseases that may
contribute to proteinuria. This is further supported by
report of Harley and Langston (2012). UPC ratios of
working dogs are affected by number of other factors such
as dietary protein content, exercise and hyperthermia
(Schaefer et al., 2007; Mustafa et al., 2011). Previous
studies reported that cage confined animals had
significantly higher UPC ratios compared to unconfined
animals, and stress in these animals would have
contributed for the difference (McCaw et al., 1985)
When we compared UPC values of dogs in the tracking
group and control groups, there was no significant
difference. This could possibly be due to the fact that they
are used for minor duties such as tracking crimes where
the chances for inhalation/ ingestion of nephrotoxic
agents are low.
There was one animal in the explosive group who was
proteinuric, but nonazotemic. According to the ACVIM
consensus statement this animal was subjected to
monitoring of kidney function. Two animals in the
narcotic and two in the explosive groups were borderline
proteinuric hence no action was taken. Therefore,
advantage of the method we established in the Diagnostic
laboratory of the VTH, would be to routinely screen dogs
for renal diseases. Nine months old puppies in the
explosive group are trained to sniff explosives such as Tri
NitroToluene (TNT), Research Department Explosive
(RDX/ Cyclonite), Dynamite, plastic explosive,
Pentaerythrite Tetranitrate (PETN), NH4NO3, Detcord,
electrical detonator, non-electrical detonator every day for
four hours in the morning and one hour in evening which
continues throughout their life time and this chronic
exposure makes them prone to renal injury.
In the narcotic group, one animal was proteinuric but
nonazotemic and two animals were borderline proteinuric.
This could possibly be due to the fact that they are trained
to sniff narcotics such as heroin, marijuana, cocain, toddy,
ganja and arrack. It has been reported from humans who
abuse narcotics are more prone to be affected with renal
failure resulting from chronic glomerulonephritis and
hypertensive nephrosclerosis (Xiqian et al., 2015).
In summary, this study assessed and established UPC
ratios in a cohort of healthy dogs in Sri Lanka. These
values were in agreement with the UPC ratios reported in
IRIS staging system. The degree of difference in UPC
values in working dogs of the Sri Lanka police warrants
attention of the authorities and clinicians regarding
clinical decisions about diagnostic, monitoring, or
therapeutic plans. Nevertheless, screening of these high
risks groups of working dogs for their kidney function and
also use of UPC measurement in other dogs as a screening
test would assist clinicians in detecting possible kidney
diseases in advance.
ACKNOWLEDGEMENT
This study was carried out with the financial support
pr ov id ed by th e Un iv er si ty Re se ar ch Gr an t
(RG/2013/18V). The authors thank Dr. M. L. A. N. R.
Deepani for help with statistical analysis of data, Ms.
M.R.C.K. Mallawa and Ms. A.H.M.W.S Aberathna for
assisting in conducting experiments and Sri Lanka Police
and Clients of dogs who volunteered to provide samples of
animals for the research project.
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S.L.Vet.J. 2016, 63:2(A): 17-21
Original article
BODY CONDITION SCORE IN LARGE PURE BRED DOGS: A
PRELIMINARY STUDY ON AGREEMENT BETWEEN OWNER'S
PERCEPTION AND SCIENTIFIC EVALUATION
M. Jagatheesan, D. D. N De Silva and H. M. H. S Ariyarathna
Department of Veterinary clinical Science, University of Peradeniya
SUMMARY: Obesity is a severe health issue among companion dogs. It may seriously impair the quality of
a dog's life predisposing to many other disease conditions. Incorrect assessment of the body condition (BC)
of their pets by the dog owners has been identified as an obstacle which would prevent early intervention in
canine obesity. This study was designed to assess the degree of agreement between the dog owner's
perception of their pet's BC and the scientific assessment of body condition score (BCS). Eighty-two large
pure bred dogs were included in the study. Scientific evaluation was performed using 1-5 BCS system
proposed by Royal Canin™ (1-emmaciated, 2- thin, 3-ideal, 4-over weight, 5- obese). Following the
scientific evaluation the owners were asked to assess their pet's BC using a grading system which included
appropriate lay terms to describe the 1-5 scoring positions in the scientific method (1=very thin, 2=thin,
3=ideal, 4=fat, 5=very fat). In addition the knowledge of necessity of feeding adjustments and exercise,
health risk of obesity and awareness of scientific BC evaluation methods were evaluated using a short
questionnaire. Agreement between the owner's perception and scientific method was analyzed using
Fleiss' kappa test. Misperception occurred among 31.71% (26/82) of the owners and there was only a fair
agreement (K=0.374321, P<0.05) between two evaluation methods. Interestingly owner misperception
about the body condition existed not only in the BCS=4/ Overweight and BCS=5/ Obese groups but also in
BCS=2 / Thin group as well. Despite the good knowledge on health risk of obesity and need of exercise
among the owners, under estimation was common in BCS=4/ Overweight (40%, 8/20) and BCS =5/ Obese
(100%, 5/5) groups which might be a sequela to owner's misperception. In the BCS=2/ Thin group there
was a prominent overestimation 3/5 (60%). Only 12.1% of the owners were aware of existence of scientific
body condition evaluation methods. Current findings are suggestive of the importance of modifying the
owner's perception about their pet's body condition and the necessity of the adequate veterinary
intervention through client communication about pet feeding and exercising.
INTRODUCTION
The prevalence of obesity in dogs has increased in
recent years (Olsen, 2011). Estimation about the incidence
of obesity in various part of the world in dogs reveals that
18%-44% of the population is obese (Anderson, 1973;
Edney & Smith, 1986; Kronfeld et al., 1991; Lund et al.,
1999; Mason, 1970; Robertson, 2003; Sonnenschein et al.,
1991). Excessive deposition of adipose tissue sequale to
positive energy balance is the cause of obesity (Colliard et
al., 2006; German, 2006; German et al., 2007; Gossellin et
al., 2007; Laflamme, 2006; McGreevy et al., 2005).
Predisposing factors of obesity include genetics,
increased daily energy intake, reduced physical activity
level, certain medications (e.g. glucocorticoids) and
diseases (e.g. hypothyroidism) (Czirják & Chereji, 2008;
German, 2006). Neutering/ spaying also suggested to be a
predisposing factor due to the effect of reduced energy
requirement (Fettman et al., 1997; Flynn et al., 1996;
Jeusette et al., 2006; Jeusette et al., 2004; Root et al.,
1996). It could deteriorate the quality of life in dogs as
well shorten the life span by predisposing to conditions
such as cardiorespiratory diseases (Ettinger & Feldman,
1995), osteoarthritis (Impellizeri et al., 2000), chronic
inflammation, diabetes mellitus (Ettinger & Feldman,
1995), and neoplasia (German, 2006; Lund et al., 1999).
There are various scientific methods used to assess the
body condition of dogs (German, 2006; Mawby et al.,
2004). They are classified into two different categories as
research techniques and clinical techniques. Research
techniques include chemical analysis (D2O),
densitometry, Dual Energy X-Ray Absorptiometry
(DEXA), ultrasound and electrical impedance. Clinical
techniques include measuring body weight and
morphometric methods (BMI: Body Mass Index, BCS:
Body Condition scoring) (German, 2006; Laflamme,
1997a; Mawby et al., 2004). Clinical methods are
comparatively quick, inexpensive and non- invasive
(German, 2006). BCS is one of the common reliable
clinical method (German et al., 2007). Four different types
of scales are in use [5 scale and 6 scale (German, 2006), 7
scale and 9 scale (Laflamme, 1997b; 1997a]. Body
condition is determined using visual assessment and
palpation which has a correlation with the degree of
subcutaneous fat deposition, abdominal fat and condition
of superficial muscles (German, 2006). Despite the
existence of many methods, the owner's perception about
the body condition of their canine companion plays a
http://doi.org/10.4038/slvj.v63i2.11
18
Jagatheesan et al
significant role to maintain an ideal BC. Some studies
conducted in UK revealed that under estimation of
BC in pet dogs is quite common among the dog
owners (Courcier et al., 2009; Eastland-Jones et al.,
2014). Research to investigate the causes of
misperception is important to maintain a healthy and
ideal dog. In Sri Lanka there are no published studies
on this topic. The objective of this study was to
assess whether there is any significant difference
between the owner's estimation and the scientific BC
evaluation (1-5 scale system, Royal CaninTM). In
addition the knowledge and attitudes on feeding
practices, necessity of exercise, awareness of health
risks of obesity and awareness of scientific body
condition evaluation methods were also assessed.
Table 1: Royal CaninTM1-5 body condition scoring system
BCS Assessment criteria
1
2
3
4
5
MATERIAL AND METHODS
The survey was carried out in the Veterinary
Teaching Hospital, Faculty of Veterinary Medicine
and Animal Science, University of Peradeniya and in
a Private Veterinary Practice in Colombo over a
period of one month. Owner's consent was obtained
prior to the assessment. Inclusion criteria for the
study were, large purebred dogs older than 6 months
(after sexual maturity) displaying breed
characteristics specified for the relevant breed (Fogle,
2002) and (RSPCA 2012). The dog owners were
requested to assess the body condition of their dog
according to the provided guidelines. First the owners
were requested to select an appropriate term out of the
five lay terms provided which they think suitable for
their pets. The used terms (very thin, thin, ideal, fat,
very fat) represented the 1 5 scoring positions in
Royal CaninTM BCS system respectively. Following
the assessment attitudes on necessity on diet
modifications for overweight/ obese dogs (Q2 in
Table 4), necessity of exercise to prevent obesity in
dogs (Q3 in Table 4), overfeeding in purebred dogs
(Q4 in Table 4), awareness of health risk of obesity
(Q5 in Table 4) and knowledge about scientific body
condition evaluation methods other than body weight
(Q6 in Table 4) were assessed using a short
questionnaire. Scientific body condition evaluation
was performed by a veterinarian using the five scale
(1-5) scoring system provided by Royal CaninTM
(Table 1). A score was determined for each dog using
visual cues followed by palpation [1=emaciated
(very thin), 2=thin, 3=ideal, 4=over weight (fat)
5=obese (very fat). In short haired dogs both the
visual assessment and palpation helped to determine
the fat deposition around the ribs, vertebral column,
abdominal tuck and waist. In long haired dogs the
determination of BCS heavily relied on palpation that
was done to ensure the fat deposition around ribs,
vertebral column, abdominal tuck and waist to
correctly determine their true body condition. The
information on gender and body weight were
collected for the purpose of record keeping. The level
Ribs, spine, pelvic bones easily visible
Obvious loss of muscle mass
No palpable fat on rib cage
Ribs, top of vertebrae, pelvic bones visible
Obvious abdominal tuck (waist)
No palpable fat on rib cage
Ribs, spine, pelvic bones not visible but easily palpable
Obvious abdominal tuck
Ribs, spine, pelvic bones palpable with difficulty
Abdominal tuck absent
Fat deposits obvious on spine base of tail
Massive fat deposits on thorax, spine and base of tail
Obvious abdominal distention
Table 2: Statistical Analysis
Kappa value
Interpretation
<0
0.01 0.20
0.21 0.40
0.41 0.60
0.61 0.80
0.81 1.00
Poor agreement
Slight agreement
Fair agreement
Moderate agreement
Substantial agreement
Almost perfect agreement
Fleiss' Kappa test interpretation
of agreement between two assessments was interpreted
according to the calculated kappa value (Fleiss' Kappa test)
using Minitab® 17 software (Table 2). To analyze the overall
agreement, a kappa value was calculated for the whole group
using the two sets of ratings produced by two assessment
methods. Agreement within the groups was assessed using the
kappa values calculated for each BCS group.
RESULTS AND DISCUSSION
A total number of 82 dogs were evaluated. Dog breeds
included were Labrador retriever, German shepherd, Golden
retrievers, Rottweiler, Doberman, Great Dane, Rhodesian
Ridgeback, Boxer, Bullmastiff, Alaskan Malamute and
Hungarian pointer. Most frequent dog breeds were Labrador
retriever (34%, n=28) and German shepherd (23%, n=19).
Overall Kappa score of 0.37 (p <0.05, 95% CI) (Landis &
Koch, 1977) indicated an overall fair agreement between
owner's assessment and scientific BCS evaluation. Owner's
assessments did not aligned with the scientific assessment in
31.7% dogs (26/82).
For further detailed analysis, dogs were divided in to five
groups based on the body condition score obtained from the
scientific method (Group 1/BCS-1, Group 2/BCS-2, Group
3/BCS-3, Group 4/BCS-4 and Group 5/BCS-5). As there were
no dogs in the Group 1/BCS 1 the particular group was not
19
Body condition score
Table 3: Overestimation, correct estimation and underestimation by the pet
owners within different groups compared to the scientific evaluation
Group
Overestimated
Group 2/ BCS 2(n=5)
Group 3/ BCS 3 (n=52)
Group 4/ BCS 4 (n=20)
Group 5/ BCS 5 (n=5)
n=3(60%)
n=6(11.5%)
n=1(5%)
n=0 (0%)
Correct
n=2(40%)
n=43(82.7%)
n=11(55%)
n=0 (0%)
Underestimated
n=3(5.8%)
n=8(40%)
n=5 (100%)
These results are formulated from the answer to the question 1 in the provided
questionnaire. There were no candidates in the Group 1/BCS 1
considered for further analysis. The agreement between
two assessments within individual groups was also
determined using the same statistical method (Fleiss's
kappa test).
Group 2/BCS=2 (n=5) had a kappa score of 0.36 (95%
CI, p<0.05) indicating a fair agreement between the two
raters. Even though there was a fair agreement between
two assessments, a considerable degree (60%, Table. 3) of
overestimation existed. However due to the small sample
size it is difficult to attempt to a conclusion that this is a
significant finding. In this group only 20% of the owners
(1/5, Table 4) believed that feed modifications are useful.
However the owner's awareness about necessity of
exercise and health risk of obesity were satisfactory (Table
4). Sixty percent (60%, Table 4) of the owners in this
group did not practice excessive feeding in purebred dogs.
Only 40% (Table 4) of owners were aware about scientific
methods of evaluating body condition in dogs.
Group 3/BCS 3 the largest group (n=52) had a kappa
score of 0.44 (95% CI, P<0.05) which indicated a
moderate agreement between the two assessments.
However, in this group also only 13.5% (Table 4) of the
owners were aware about scientific evaluation methods
for assessment of body condition in dogs. Even though
only 13.5% (Table. 4) of the owners were aware about
scientific body condition evaluation methods, 90.4%
(Table. 4) of them were aware about the health risk of
obesity and necessity of exercise (Table. 4).
A
considerable proportion (40.4%, Table. 4) of owners
strictly rejected excess feeding in purebred dogs. This
might have played an important role in maintaining an
optimum BCS even in the absence of knowledge about a
scientific BCS evaluation method. Moreover, the owner's
awareness on necessity of exercise and the awareness on
the health risk of obesity might have contributed
substantially to maintaining these dogs in ideal body
condition. However, 34.6% dogs had been overfed in this
group which might be due to the lack of knowledge about a
scientific evaluation method.
Group 4/BCS 4 (n=20) carried a kappa score of 0.38
(95%CI, P<0.05) which indicated a fair agreement
between the two assessments. All dogs in this group were
overweight according to the scientific evaluation .
Interestingly majority of owners in this group (95%, Table
4) were aware of the health risks of obesity. Moreover
80% (Table 4) of the dog owners accepted that exercise is
important. However only 5% (Table. 4) of the dog owners
were aware about an existence of scientific body condition
evaluation methods other than body weight. Therefore,
had they provided with a correct body condition scores
assessed following a scientific method, they might have
attempted to modify feeding practices and exercise to
reduce the risk of being overweight. However, in spite of
the awareness of the risk of obesity and necessity of
exercise, 40% (Table 4) of the dog owners believed that
purebred dogs need excessive feeding and only 35%
(Table 4) of the owners accepted the need to modify the
feed. Therefore, it is necessary to educate the pet owners
that feed requirement in dogs does not depend on the breed
but according to the energy expenditure and physiological
status of the dog.
Group 5/BCS=5, n=5) had a kappa score of -0.037
(95%CI p >0.05) indicating no agreement between the two
methods of assessment. According to the scientific
evaluation all the dogs in this group were obese and
dietary intervention and exercising is essential as they are
already predisposed to various health issues (pressure
wounds n = 3, exercise intolerance n = 4, infertility = 1).
Interestingly under estimation by the pet owners was
100% (Table. 4) even though all the owners were aware of
the health risk of obesity as well as the need for exercise.
Sixty percent 60% (Table 4) of the owners were willing to
modify feed. Excessive feeding of purebred dogs was
practiced by 40% (Table 4) of the owners while 40%
(Table 4) of the owners disagreed with that. None of the
dog owners in this group were aware of scientific body
condition evaluation methods other than body weight
(Table 4). Therefore, misperception of the body condition
of their pets might be an important contributory factor for
obesity in this group. However only 5 dogs were in this
study and it is necessary to have a larger group to identify
the consistency and significance of these findings.
In the groups 4 and 5 despite the awareness of health
risks and necessity of exercise, dogs were overweight/
obese. One possible explanation for this is existence of
underlying disease conditions such as hypothyroidism,
cardiac diseases and arthritis, which may predispose the
dogs to obesity or interfere with exercising. However, the
investigation of these possible underlying factors is
beyond the scope of this study. Neutering also known to
20%
(n=1)
80%
(n=4)
0%
(n=0)
0%
(n=0)
0%
(n=0)
100%
(n=5)
20%
(n=1)
40%
(n=2)
5%
(n=1)
90%
(n=18)
5%
(n=1)
0%
(n=0)
5%
(n=1)
95%
(n=1)
30%
(n=6)
30%
(n=6)
17.3%
(n=9)
69.2%
(n=36)
13.5%
(n=7)
1.9%
(n=1)
7.7%
(n=4)
90.4%
(n=47)
25%
(n=13)
40.4%
(n=21)
20%
(n=1)
40%
(n=2)
40%
(n=2)
0%
(n=0)
40%
(n=2)
60%
(n=3)
0%
(n=0)
60%
(n=3)
DK
N
Y
DK
N
Y
DK
N
cause changes in the basal metabolism due to hormonal
alterations predisposing dogs to obesity. However, as all the
dogs included in Group 4 and 5 are reported to be intact this
argument is least valid for these groups.
In a previous study conducted in UK (Courcier, et al. 2009)
44.12% (95%CI: 40.39-47.85), n=300) of owners
misperceived the body condition of their dogs. Out of these
owners 77% (n=231) of the owners under estimated the body
condition of their dogs. Fifty-three percent of overweight dog
owners had under estimated their dogs (Courcier, et al. 2009).
Our study demonstrated an overall 21.9% underestimation
(18/82, Table 2) which is comparatively a low difference.
However the underestimation percentage within the Obese/
Overweight groups was 52% (13/25, Table 2). Interestingly
in our study out of the misperceived owners (n=26), 38.5%
(10/26, Table 2) over estimated their dogs which was not
observed in the UK study.
CONCLUSIONS
Results of this study reveals that even though some of the
pet owners are aware about the health risks of obesity they do
not attempt for necessary diet modification and/or exercise
due to improper perception of their pet's body condition.
Therefore, Current findings indicate the importance of
veterinary intervention to assess the body condition using a
scientific method (based on standard criteria) in pure bred pet
dogs to minimize obesity related health risks.
Y
40%
(n=2)
34.6%
(n=18)
40%
(n=8)
40%
(n=2)
DK
0%
(n=0)
1.9%
(n=1)
5%
(n=1)
0%
(n=0)
N
0%
(n=0)
7.7%
(n=4)
15%
(n=3)
0%
(n=0)
Y
100%
(n=5)
90.4%
(n=47)
80%
(n=16)
100%
(n=5)
DK
20%
(n=1)
15.4%
(n=8)
5%
(n=1)
0%
(n=0)
N
60%
(n=3)
61.5%
(n=32)
60%
(n=12)
40%
(n=2)
Y
20%
(n=1)
23.1%
(n=12)
35%
(n=7)
60%
(n=3)
BCS-2
(n=5)
BCS-3
(n=52)
BCS-4
(n=20)
BCS-5
(n=5)
ACKNOWLEDGEMENT
Group
Q3
Q4
Q5
Q6
Jagatheesan et al
Q2
Table 4: Attitudes and knowledge on feeding practices, necessity of exercise, obesity and scientific methods of evaluating body condition in dogs
20
Authors would like to acknowledge the staff of Pet Vet
Clinic, 421/5, Malalasekara Mawatha, Colombo 07 for their
assistance with data collection.
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Clinical communication
FELINE PANLEUKOPENIA VIRUS INFECTION IN A
CAPTIVE-BRED BENGAL TIGER (Panther tigris tigris) AND
A LEOPARD (Panthera pradus)
D. R. A. Dissanayake1, I. D. Silva1, Sashikala Gamage1, Dinuka Sonnadara2, M. R. B. N.
3
3
1
4
Bandara , S. S. Alokabandara , V.P. Priyanka Jayapani and Waruni Jayaweera
1
Veterinary Teaching Hospital, Department of Veterinary Clinical Sciences, Faculty of
Veterinary Medicine and Animal Science, University of Peradeniya, Peradeniya
2
National Zoological Garden, Dehiwala, 3National Zoological Garden, Pinnawala
4
Department of Veterinary Pathobiology, Faculty of Veterinary Medicine Animal Science
INTRODUCTION
Feline panleukopenia (FPL) (also known as feline
distemper) is a contagious and fatal disease of both
domesticated and wild felids. The disease is caused by
Feline panleukopenia virus (FPLV) of the family parvo
viridae. This virus is closely related to the canine parvo
virus (CPV), mink enteritis virus (MEV) and raccoon
parvo virus and only show <2 % difference in their
genome (Steinel et al., 2001; Tailor et al., 1999). In vitro
culture and in vivo inoculation of virus of the family
parvoviridae have confirmed overlapping host ranges, and
their classification is mostly based on the host from which
the virus has been isolated (Shackelton et al., 2005;
Truyen and Parrish, 1992). The FPLV among wild felids
are less commonly reported due to difficulties in
observing clinical signs, although, many evidences
confirm that FPLV can affect almost all species of wild
felids (Lane et al., 2016; Olmsted et al., 1992).
The Feline panleukopenia is transmitted through
oronasal route from contaminated food, fomites and air.
Viral multiplication in actively dividing cells (such as,
bone marrow, lymphoid tissue, intestinal epithelium,
cerebellum and retina of neonate kittens) cause lysis of
cells resulting in panleukopenia, diarrhoea, ataxia,
incoordination and vision impairment in young animals
(Steinel et al., 2001; Truyen et al., 2009; Parrish, 1995),
and abortion, still birth and fetal resorption in infected
pregnant queens (Jordan and Sever, 1994).
Recently, a leopard cub (Panthera pardus) and a
Bengal tiger cub (Panthera tigris tigris ) housed at two
distantly located Zoological Gardens in Sri Lanka had
developed haemorrhagic diarrhoea, and the aetiology was
confirmed as FPLV by polymerase chain reaction (PCR)
and sequencing. This communication discusses the
clinical presentations, and pathological changes observed
in these two felids due to FPLV.
Case 1: A ten-month old unvaccinated female leopard cub
housed at Pinnawala open zoo had developed lethargy,
acute inappetance, haemorrhagic diarrhoea and vomiting
in mid September 2016. Blood on the first day of the
clinical onset revealed a total leukocyte count (WBC) of
22.47×103/µl, without significant changes in the
differential counts other than the presence of 4% of band
neutrophils. The hematocrit was 35.3% and she was
moderately dehydrated at the time of blood sampling for
laboratory analysis. Differential diagnoses included,
enteritis caused by CPV, FPLV, Salmonella enterica or E.
coli (enterohaemorrhagic E. coli). The faecal sample was
negative for Salmonella enterica. The PCR amplifying the
VP2 gene of parvo virus, (F 5'TTACTAAGAACAGGTGATGAA-3' and R- 5'ATTTGGATAAACTGGTGGT-3') confirmed parvo viral
DNA in faeces (Figure 1).
The leopard cub was housed in isolation and treated
with ceftriaxone (50 mg/ Kg BW, IV, 12 h), metranidazole
(20 mg/ Kg BW, IV,12h ), metoclopramide (0.5 mg/ Kg
BW, IV, 24h), catosal (1 ml containing, 100mg of 1-[nButylamino]-1- methylethyl phosphonous acid
[Butaphosphan] and 0.05mg of vitamin B12). In addition,
intravenous fluids (0.9% NaCl and lactated ringers) were
administered to correct dehydration and electrolyte loss,
and 25% dextrose and amino acids were administered as
energy and protein supplements. No oral food was
introduced until vomiting ceased. She responded well to
treatment and recovered after 4 days.
Case 2: A three month old unvaccinated female Bengal
tiger cub housed at National Zoological Gardens,
Dehiwala developed haemorrhagic diarrhoea and
vomiting. The faecal sample revealed eggs of
Ancylostoma species, and hence she was treated with an
oral antihelminthic (Drontal plus at 10 mg/ Kg BW). Fluid
therapy could not be given due to difficulties in separating
the cub from the dam, and the tiger cub died the following
day.
Necropsy revealed a dehydrated carcass with severe
hyperaemia on the serosae of the duodenum and jejunum,
and petechial haemorrhages on the rest of the intestine.
The dilated intestines contained haemorrhagic watery
feces which tested positive for parvo viral DNA (Figure 1),
http://doi.org/10.4038/slvj.v63i2.12
24
Dissanayake et al
Figure 1: Agarose gel electrophoresis of
amplified PCR products of VP2 gene. MM:
100bp molecular ladder, Le: Leopard-faecal
sample, BT: Bengal Tiger-faecal sample, +ve:
positive control and ve: negative control
Figure 2 a. H & E stained section of the
duodenum (10X10) of Bengal tiger infected with
FPLV. Loss of epithelium of intestinal crypt
(long arrows) and destruction of intestinal villi
(short arrows)
but not for Salmonella. Mesenteric lymph nodes and
spleen were enlarged. Histopathology revealed
shortening and collapsing of intestinal villi with extensive
loss of epithelium in the intestinal crypts. In addition,
(Figure 2a and 2b) marked bacterial colonization was seen
in the mucosae and sub mucosae of the intestines (Figure
2b).
Figure 2 b. H & E stained section of the
duodenum of Bengal tiger infected with FPLV
(10 X10 0). Des tru cti on of epi the liu m of
intestinal crypt (arrow heads) and bacterial
colonization in the mucosae (long arrows)
DISCUSSION
In addition to domestic and wild felids, FPLV can also
affects racoon, mink and fox (Truyen et al., 2009). It is
important to note that the FPLV is not the only parvo virus
affecting cats, and new variants of canine parvo virus
(CPV2a and b) are also capable of causing haemorrhagic
Feline panleukopenia virus infection
enteritis in both domestic and wild cats (Truyen et al.,
1996). Passaging of FPLV and CPV2 through multiple
hosts has caused mutations in the gene encoding viral
capsid. Resulting changes in the capsid has broaden the
host range of the virus (Truyen et al., 2009).
In order to identify the species of the parvo virus, PCR
products were sequenced. The PCR assay used in the
present study can screen for both CPV and FPLV. The
resulting PCR amplicons were sequenced for species
identification. Sequences were compared using BLASTn
against the GenBank (http://www.ncbi.nlm.nih.gov/)
non-redundant nucleotide collection database to identity
the closest match. The results confirmed that the two
sequences were identical and highly similar (96%
coverage and 98% identity) to the Feline panleukopenia
virus strain 42/06-G8, isolated from a cat with FPL in Italy
(gene bank accession EU498699.1). Though many
clinical cases with signs similar to FPL were observed
among domestic cats in Sri Lanka, none of those cases
were confirmed as FPLV by molecular or any other
diagnostic methods. Therefore we are unable to confirm
that the FPLV strain in wild felines is similar to the FPLV
circulating among domestic cats in Sri Lanka.
Previous reports have shown that the FPL may develop
as a clinical or sub clinical infection. Severity of the
infection may depend on various factors including
maternal derived antibodies, age, the immune status of the
individual and underlying diseases. The concurrent worm
infestation could have contributed to the severity of the
infection in the Bengal tiger. Both Ancylostoma and FPLV
damage the intestinal epithelium facilitating the intestinal
microbes to translocate to blood and various organs
causing septicaemia and endotoxaemia. In a previous
study, a germ free cat had developed only mild transient
infection following FPLV infections (Carlson et al., 1977),
indicating that the systemic entry of intestinal bacteria
determine the severity and the prognosis of FPLV
infection. Furthermore, the young age of the animal (<3
months) would have contributed to the severe outcome.
Previous reports have shown FPLV infected kittens to
have a very high (90%) mortality (Truyen et al., 2009)
Histopathology of the tiger cub showed epithelial cell
destruction mainly in the intestinal crypts. Cells in crypts
of Lieberkuhn have high replication rate and the FPLV
initially start to grow in these cells. With the progression
of the infection, villi atrophy occur due to absence of new
cells to replace the old epithelium (Goddard and Leisewitz,
2010). The most common symptoms of FPL; ie., diarrhoea,
vomiting and dehydration occur due to damaged
absorptive surface of the intestines.
Since there is no specific treatment available for FPL
infection, affected animals should be managed and treated
according to the symptoms. A broad spectrum
antimicrobial, antiemetic and appropriate fluid and energy
corrections should be included in the treatment and
management plan to avoid complications due to
septicaemia, endotoxaemia or severe dehydration. It is
also vital to isolate the suspected patients as FPL is highly
25
contagious. When FPL infection is suspected in animals
in captivity, extra precautions should be taken to prevent
the spread of the infection. In order to prevent FPL in
Zoological Gardens, all members of the cat family should
be properly vaccinated. Three vaccines, at 8-9 weeks, 1112 weeks and 16-20 weeks of age are recommended for
kittens when there is a high infection pressure. A booster
vaccine should be given, when animal is one year old
followed by boosters at intervals of three year or more.
ACKNOWLEDGEMENT
Authors would like to thank Ayoma Hettiarachchi, K.
Thanajayan, Chandima Mallawa, Ushantha Hewagamage
for their support.
REFERENCES
Carlson, J., Scott, F., Duncan, J. (1977). Feline
panleukopenia I. Pathogenesis in germfree and specific
pathogen-free cats. Veterinary Pathology Online, 14,
79-88. https://doi.org/10.1177/030098587701400110
Goddard, A., Leisewitz, A.L. (2010). Canine parvovirus.
Veterinary Clinics of North America: Small Animal
Practice, 40, 1041-1053.
https://doi.org/10.1016/j.cvsm.2010.07.007
Jordan, E.K., Sever, J.L. (1994). Fetal damage caused by
parvoviral infections. Reproductive Toxicology, 8, 161189. https://doi.org/10.1016/0890-6238(94)90023-X
Lane, E.P., Brettschneider, H., Caldwell, P., Oosthuizen, A.,
Dalton, D.L., du Plessis, L., Steyl, J., Kotze, A. (2016).
Feline panleukopaenia virus in captive non-domestic
felids in South Africa. Onderstepoort Journal of
Veterinary Research, 83, 8.
https://doi.org/10.4102/ojvr.v83i1.1099
Olmsted, R.A., Langley, R., Roelke, M.E., Goeken, R.M.,
Adger-Johnson, D., Goff, J.P., Albert, J.P., Packer, C.,
Laurenson, M.K., Caro, T.M. (1992). Worldwide
prevalence of lentivirus infection in wild feline species:
epidemiologic and phylogenetic aspects. Journal of
Virology, 66, 6008-6018
Parrish, C.R. (1995). 3 Pathogenesis of feline
panleukopenia virus and canine parvovirus. Baillière's
clinical haematology. 8, 57-71.
https://doi.org/10.1016/S0950-3536(05)80232-X
Shackelton, L.A., Parrish, C.R., Truyen, U., Holmes, E.C.
(2005). High rate of viral evolution associated with the
emergence of carnivore parvovirus. Proceedings of the
National Academy of Sciences of the United States of
America, 102, 379- 384.
https://doi.org/10.1073/pnas.0406765102
Steinel, A., Parrish, C.R., Bloom, M.E., Truyen, U. (2001).
Parvovirus infections in wild carnivores. Journal of
Wildlife Diseases, 37, 594-607.
https://doi.org/10.7589/0090-3558-37.3.594
Tailor, C.S., Willett, B.J., Kabat, D. (1999). A putative cell
surface receptor for anemia-inducing feline leukemia
virus subgroup C is a member of a transporter
superfamily. Journal of virology, 73, 6500-6505
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Truyen, U., Addie, D., Belák, S., Boucraut-Baralon, C.,
Egberink, H., Frymus, T., Gruffydd-Jones, T.,
Hartmann, K., Hosie, M.J., Lloret, A. (2009). Feline
panleukopenia. ABCD guidelines on prevention and
management. Journal of Feline Medicine & Surgery, 11,
538-546. https://doi.org/10.1016/j.jfms.2009.05.002
Truyen, U., Evermann, J.F., Vieller, E., Parrish, C.R.
(1996). Evolution of canine parvovirus involved loss
and gain of feline host range. Virology, 215, 186-189.
https://doi.org/10.1006/viro.1996.0021
Truyen, U., Parrish, C. (1992). Canine and feline host
ranges of canine parvovirus and feline panleukopenia
virus: distinct host cell tropisms of each virus in vitro
and in vivo. Journal of virology, 66, 5399-5408
S.L.Vet.J. 2016, 63:2(A): 27-30
Clinical communication
PREVALENCE, TREATMENT AND MANAGEMENT OF HEALTH
CONDITIONS IN PET RABBITS PRESENTED TO THE VETERINARY
TEACHING HOSPITAL (VTH), UNIVERSITY OF PERADENIYA
K.K. Sumanasekera, M.G.C.M. Jayasinghe, K.G. Indika,
R.E.H. Perera and D.D.N. de Silva
Veterinary Teaching Hospital, Department of Veterinary Clinical Sciences, Faculty of Veterinary Medicine and
Animal Science, University of Peradeniya
SUMMARY: Forty five pet rabbits of different breeds with various clinical conditions were presented to
the VTH from January 2014 to March 2016 reflecting an emerging trend in companion animal ownership
and health concerns. Of these cases, Psoroptic mange was the commonest disease condition while
traumatic injuries (26.7%) including dog bite wounds, fractures, eye conditions were regularly
encountered. Chemical trauma and infectious diseases (17.8%) namely, ulcerative pododermatitis,
respiratory tract and urinary tract infections; miscellaneous cases such as floppy rabbit syndrome, heat
stress, nutritional deficiency and prolapsed vagina were also presented. The objective of this
communication is to make the veterinary practitioners aware of the common health conditions
encountered in pet rabbits and how those were treated and managed.
BACKGROUND
Domestic rabbits are descended from the European
rabbit, which belongs to the order Lagomorpha.
Numerous breeds of domestic rabbits have been
developed with various characteristics. Common rabbit
breeds in Sri Lanka included New Zealand white, Flemish
giant, Californian white and Chinchilla. Pet rabbits are
mostly cross bred of these original breeds. Forty five pet
rabbits of different breeds (pure and cross bred) with
various clinical complications presented to the VTH from
January 2014 to March 2016 are discussed here.
Psoroptic mange is a common mite infestation of rabbits
caused by Psoroptes cuniculi. This condition was
encountered in 19 out of 45 rabbits and clinical
presentation of most rabbits was otitis externa with crusty
exudates that form within the ear canal and extend up to
the pinna and also the skin form on the nose (Figure 1),
paws and the perineal region with extremely pruritic
lesions. Diagnosis was confirmed by microscopic
examination of the exudates; mites were also visible to the
naked eye. These were treated with Ivermectin (0.4mg/kg)
subcutaneously and followed-up after ten to twelve days.
All the cases responded well to this treatment.
Among the cases presented 20% were traumatic injuries
mostly due to dog bites and management related injuries.
First case was an extensive infected laceration due to a dog
bite at the base of the left ear up to the neck (Figure 2). The
wound was cleaned with normal saline, dressed with
Povidone Iodine solution and enrofloxacin (10mg/kg) sid
SC was administered. There was evidence of healing of
the wound with daily treatment but it succumbed to the
condition after three days. The other rabbit aged five
months was presented with a dog bite of the left hind limb,
where the tibia fibula, meta tarsal and digital bones with
the associated soft tissue were lost resulting in an avulsion.
Patient was stabilized with intravenous fluid (0.9% saline
+ 10% dextrose) and analgesics (meloxicam 0.2mg/kg)
PO. Amputation of the left hind limb was performed under
general anaesthesia (GA); induction with ketamine HCl
(35mg/kg) and xylazine HCl (5mg/kg) IM. Enrofloxacin
(10mg/ kg) was admini stered to preven t bacter ial
infections. Even though the patient showed difficulty in
hopping there was uneventful and complete recovery.
Four rabbits were presente d with manageme nt/
handling issues causing fractures and one with epistaxis. A
cross bred rabbit with closed complete right tibia-fibula
fracture was treated using intra-medullary pin insertion.
Pin was removed fourteen days later and the patient
showed a good response. A five month old rabbit with
complete closed mid tibia-fibula fracture was treated by
external coaptation using plaster of paris (POP), under GA.
Owners were advised to reduce movements and the POP
was removed ten days later. Another three- year old rabbit
was reported with epistaxis after falling down from a
height, and was managed using cold fomentation,
application of one drop of Oxymetazoline® 3 times a day
for 3 days into each nostril which resulted in complete
recovery. A 6- month old rabbit presented paraplegic after
fallen down while carrying by the owner. Pain sensation
was present in the caudal region of the body, but no
fractures were detected in the radiograph. Pain
management was done using meloxicam orally (0.2mg/kg
sid) for five days and the owner was advised to do
physiotherapy (hot fomentation) but had a poor prognosis.
Two rabbits with hypersensitivity reaction due to skin
exposure to Lysol® were presented and treated with
dexamethasone (2mg/ kg) and chlorpheniramine maleate
http://doi.org/10.4038/slvj.v63i2.13
28
Sumanasekera et al
Figure 1: Psoroptic mange - skin
form
Figure 2: Infected laceration
Figure 3: Abscess in the eye
Figure 4: Ulcerative pododermatitis
Figure 5: Sludgy urine
Figure 6: Forward head tilt
Figure 7: Abscess below the mandible
(0.4mg/kg) IM after profusely cleansing off the chemical with
water. Traumatic injuries due to unknown causes were also
reported, such as paraplegia and scrotal laceration.
Ocular conditions of rabbits were also encountered. A
rabbit with an eye abscess (Figure 3) was treated with
enucleation under GA through lateral canthotomy followed
by removal of the globe and tarsorrhaphy. Corneal opacity
and conjunctivitis in an adult rabbit were treated with
ciprofloxacin (1 drop bid) and betamethasone (1 drop tid) eye
drops for fourteen days which resulted in complete recovery.
A rabbit with corneal ulcer was treated with Keterolac®(1 drop
tid) and ciprofloxacin (1 drop bid) eye drops for fourteen days
but the response was unsatisfactory and thus enucleation had
to be performed.
Two cases with ulcerative pododermatitis were reported
(Figure 4). They were anorexic and the lesions were
manifested as ulcerated infected areas of skin/ abscess on the
caudal aspect of the tarsus and metatarsus. The full blood
counts were unremarkable. Abscess at the hock was drained
29
Health conditions in pet rabbits
off and a bandage was applied af ter applying
Soframycin® ointment and in addition enrofloxacin
and meloxicam were also administered SC. Owner was
advised to change the bandage regularly and also to
apply Soframycin® ointment and change the bedding
material (hay) regularly.
One rabbit was presented with pododermatitis with
concomitant psoroptic mange and was treated with
ivermectin and enrofloxacin SC which resulted in good
recovery.
Three rabbits were presented with respiratory tract
infection. The clinical signs included loss of appetite,
clear nasal discharges, shallow rapid breathing and
wheezing. Treatment with Cotrimoxazole® (30mg/kg
bid) and dexamethasone (1mg/kg) for five days gave a
positive response, and the treatment protocol was
similar to that of protocol proposed by Longley, 2010.
Nasal swab of one of the rabbits was positive for
Pseudomonas and Staphylococcus. The antibiotic
susceptibility test indicated that the bacteria were
susceptible to enrofloxacin, cotrimoxazole and
ciprofloxacin.
A rabbit with a complaint of dysuria for four days,
dribbling of urine, abdominal pain and reduced appetite
was presented. Urinanalysis revealed high calcium
carbonate crystaluria, pyuria, haematuria and pH of 9
while lateral abdominal radiograph revealed urine
filled distended bladder with radio opaque sediment.
The condition was diagnosed as sludgy urine and
cystitis (Figure 5). Urine was removed from the bladder
manually under sedation (diazepam 1 mg/ kg) IV and
treated with enrofloxacin (10mg/kg SC sid),
Furosemide® (1 mg/ kg IV sid), meloxicam (0.2 mg/ kg
SC sid) and vitamin C (25 mg PO sid) (Carpenter, 2005)
for three days. The owner was advised to supply ad
libitum drinking water and change the diet to fruits and
vegetables with high water content (Kestenman, 2016).
The patient responded well to the treatment.
Two similar cases were presented with forward head
tilt, inability to lift the head or move the fore limbs, mild
dehydration and reduced appetite. Based on the clinical
signs, the condition was tentatively diagnosed as floppy
rabbit syndrome (Figure 6) and SC fluid therapy,
enrofloxacin (10mg/kg) and dexamethasone (1mg/kg)
IM were administered. Ciprofloxacin (10 mg/ kg) and
dexamethasone were prescribed orally for two more
days. Both rabbits recovered completely.
Two cases of abscesses were present with white thick
creamy content; both were present below the mandible
(Figure 7). The culture was positive for Pasteurella.
Abscesses were surgically drained under GA and
enrofloxacin SC was given as antibiotic therapy. It has
been reported that chronic suppuration is common in
rabbits and slow growing, well-encapsulated, relatively
painless abscesses develop readily. Pasteurella
multocida has been frequently isolated from these
abscesses (Frances Harcourt-Brown, 2002).
There were other cases reported such as vaginal
prolapse (which succumbed to the condition few
minutes after admission) reproductive failure due to
malnutrition which was diagnosed based on the condition of
the body. The condition of two rabbits presented with
reduced appetite could not be diagnosed while another adult
rabbit was diagnosed having heat stress which presented with
respiratory distress, elevated rectal temperature (1060f) and
sudden collapse. It recovered after giving intravenous fluid
and keeping in a quiet air conditioned room.
DISCUSSION AND CLINICAL SIGNIFICANCE
All rabbits infested with psoroptes cuniculi were similar to
typical psoroptic mange cases reported in literature (Frances
Harcourt-Brown, 2002). Information on the life cycle of the
parasite is important to decide the treatment plan; eggs hatch
after four days, but the mite can survive in the environment
up to 21 days (medirabbit.com, 2016; Urquhart, 1996).
Therefore, follow-ups are necessary. Owners should be
advised not to remove the crust as it may cause severe pain
and may also leads to wounds. This infestation can rapidly
spread among rabbits (Frances Harcourt-Brown, 2002).
Therefore all the rabbits of the group should be closely
examined when one is infested.
Table 1: Summary of the health conditions in rabbits
reported in this study
Condition
Psoroptic mange
Ulcerative pododermatitis
Traumatic injuries
Ocular conditions
Abscesses
Respiratory tract infections
Floppy rabbit syndrome
Sludgy urine
Heat stress
Nutritional deficiency
Prolapsed vagina
Undiagnosed
Number of cases Percentage
19/45
2/45
9/45
3/45
2/45
3/45
2/45
1/45
1/45
1/45
1/45
2/45
42.22
4.44
20.00
6.67
4.44
6.67
4.44
2.22
2.22
2.22
2.22
4.44
Ulcerative pododermatitis is a very painful and serious
condition which is difficult to cure once sets in. It has been
recognized as a secondary disease to physical,
conformational or husbandry problem (Frances HarcourtBrown, 2002). The most common bacteria isolated from
these lesions are Staphylococcus aureus and Pasteurella
multocida (Frances Harcourt-Brown, 2002).
Floppy rabbit syndrome is a condition causing generalised
muscular weakness. Aetiology of this disease is not known
although several possibilities exist, such as hypokalaemia,
nutritional muscular dystrophy, myasthenia gravis and spinal
cord disease. (Esther van Praag, medirabbit.com, 2016). A
feature of this condition is that the recovery can take place
with supportive care.
30
Sumanasekera et al
There is a high probability of rabbits acquiring psoriatic
mange and subjected to traumatic injuries similar to the
reports by Okerman, (1994). Therefore, when rearing
rabbits, owners should take measures to reduce traumatic
injuries such as constructing cages with a suitable floor
which facilitate drainage but prevent accidental trapping
of limbs; and also to provide them with proper nutrition.
Mixed infection was reported only in a one case where the
rabbit was having both psoriatic mange and ulcerative
pododermatitis. As rabbits are animals that are prone to
stress easily and thus gentle handling with minimum
stress is very important in disease diagnosis and treating
by the veterinarians and owners.
ACKNOWLEDGEMENT
Authors extend their appreciation to the academic and
non-academic staff of the VTH for their enormous
cooperation extended during this study. The dedication of
the final year veterinary students of the batches 2009/2010
and 2010/2011, Faculty of Veterinary Medicine and
Animal Science, University of Peradeniya is especially
acknowledged in managing the cases.
REFERENCES
Carpenter, J. W. (2005). Exotic animal formulary. 1st ed.
Elsevier Saunders, 411-438.
Esther van Praag. Ear mite: Psoroptes cuniculi.
Medirabbit. 2003. http://meddirabbit.html, [accessed 7
April 2016]
Esther van Praag. Floppy rabbit syndrome- general
muscle weakness. Medirabbit. 2003.
http://meddirabbit.html, [accessed 7 Apr. 2016]
Frances Harcourt-Brown. (2002). Textbook of Rabbit
Medicine. 1st ed. Butterworth-Heinemann, 52-163, 206248, 292-351, 361-385.
Kestenman, S.A. Bladder disease and bladder stones in
the rabbit. House rabbit society. http://rabbit.org
[accessed 16 Apr.2016]
Longley, L. (2010). Small animal exotic pet medicine. 1st
ed. Saunders Elsevier, 45-92.
Okerman, L. (1994). Diseases of domestic rabbits, 1st ed.
Blackwell scientific publications.
Urquhart, G.M., Armour J., Duncan, J.L., Dunn, A.M.,
Jennings, F.W. (1996) Veterinary Parasitology, 2nd
ed.Blackwell publications, 197-200.
S.L.Vet.J. 2016, 63:2(B): 31-34
FOOD SAFETY MONITORING AND
SURVEILLANCE SYSTEM FOR SRI LANKA
THE GLOBAL PERSPECTIVE ON FOOD
SAFETY AND FOOD BORNE ILLNESS
Food safety creates major public health issues
worldwide. Food borne illnesses are common, costly, and
a so far preventable public health problem. The food borne
illnesses are defined as diseases, usually either infectious
or toxic in nature, caused by agents that enter the body
through the ingestion of food or water (WHO: Fact sheet
N°237, 2007). These contaminations in food and water
can create an enormous social and economic burden on a
society and their health system in both developed and
developing countries such as Sri Lanka. When foodborne
illness is considered both biological and chemical agents
are involved. The estimated reported global burden of
foodborne illness is caused by 31 bacteria, viruses,
parasites, toxins and chemicals (WHO, 2016). The
commonly identified biological agents are
Campylobacter spp., E.coli O157:H7, Listeria
monocytogenes, certain serotypes of Salmonella enterica,
Shigella spp, Clostridium botulinum, Staphylococcus
aureus, Norovirus, Toxoplasma gondii and foodborne
trematodiases (WHO: Fact sheet, 2016).
It has been reported that globally there are nearly 1.7
billion cases of diarrheal disease every year and this kills
around 760 000 children under the age of five years
(WHO: Fact sheet N°330, 2013). A great proportion of
these cases can be attributed to contamination of food and
drinking water. Developing countries such as Sri Lanka
also bears the brunt of the problem due to the presence of a
wide range of foodborne diseases.
PREVAILING STUDIES AND KNOWLEDGE GAP
ON FOOD BORNE PATHOGENS AND ITS
IMPACT
Passive surveillance studies have being carried out in
Sri Lanka regarding the identification of food borne
pathogens. In Sri Lanka most frequent were diarrheal
diseases caused by Campylobacter spp., and Norovirus. In
2012, a total of 4003 clinically confirmed human cases of
foodborne illnesses were reported. Among those, the
majority were young children below 9 years of age
(Annual Epidemiological Bulletin, 2012). In a study
conducted in 2012 it has been found that the prevalence of
Bacillus cereus was 56 % in Chinese style fried rice that
was available in Colombo city. The organism was
responsible for causing the symptoms of diarrhea and
vomiting. The identified risk factors in that study were
storage of boiled rice at room temperature for more than
forty eight hours and the cooking frequency (Perera and
Ranasinghe, 2012). Toxigenic Aspergillus species,
Aflatoxin B1, Aflatoxin G1 have been also identified in
parboiled rice and raw milled rice due to the increased
storage time and the storage method (Bandara et al., 1991a,
Bandara et al., 1991b). Ocratoxin has been identified in
the consumable food samples in the North Central
Province of the country (Wanigasuriya et al., 2008).
Fish collected from the Negombo area and distributed in
suburbs of Colombo had been positive for the presence of
Escherichia coli, Salmonella spp and Listeria
monocytogenes along the different steps of the supply
chain such as at the boat, the ice manufacturing plant and
at post-harvest handling. Fish is important in the context
of contamination as it is easily perishable (Ariyawansa et
al., 2016). Salmonella species have been identified in both
captured and cultured shrimp at a prevalence of 12.8 % in
a study conducted in 2008. A Salmonella infection could
results in nausea, vomiting and abdominal cramps
(Kamalika et al., 2008). In dried fish from the markets in
Kandy area the presence of different fungal isolates
including Aspergillus niger and Aspergillus flavus was
shown (Atapattu and Samarajeewa, 1990). Traces of
heavy metals such as Ca, Cd, Cu, Fe, Hg, K, Mg, Mn, Na, P,
Sr and Zn have also been found in Tilapia reared in the
water reser voirs . The stand ards were below the
international recommended levels during the year 2009
(Allinson et al., 2009).
Rajasooriya et al., (2002) have conducted a study on
ground water quality at one region of Valigamam and
identified a large proportion of wells having high level of
nitrate which exceed the WHO standard due to intensive
agriculture practices through high inputs of artificial and
natural fertilizers. However, this water is used for different
farming practices and also in the farmers' houses. The
possible contamination of fecal coliforms in drinking
water resources in Jaffna area has been documented
(Muralithas et al., 2011).
Foodborne illnesses are of major concern in the YOPI
(Young, Old , Pregnant and Immunosuppressed) group of
people because of the reduced immunity levels (Havelaar
et al., 2015, Barbuddhe et al., 2012, Kirk et al., 2015). In
2014 the annual health bulletin of Sri Lanka had indicated
1072 cases of typhoid fever were reported and 27.6 % was
found in the age group of 5-14 years old. The number of
dysentery patients was 4832 cases (33.7% were children
1-4 years) and 2056 viral hepatitis cases (27.6% were 5-14
years).
Umanga C. Gunasekera1, 2, Mahalingam Muralithas1, 2 and Maximilian P.O. Baumann2
Department of Animal Production and Health, Sri Lanka; 2Postgraduate Studies in International Animal Health,
Department of Veterinary Medicine, Freie Universitat Berlin, Germany
http://doi.org/10.4038/slvj.v63i2.14
1
32
Gunasekera et al
Some pathogens that are reported from food borne
illness cases emerge as multi drug resistance type and this
cause further challenge to control the disease (Garedew et
al., 2015; Routh et al., 2015; Liu et al., 2013).
Continuous screening and monitoring of antimicrobial
susceptibility in bacteria from any animal or plant
originated food is extremely important to ensure the
betterment of global public health and economy as well.
The Food Microbiology Laboratory at the Department of
Veterinary Public Health & Pharmacology, Faculty of
Veterinary Medicine & Animal Sciences, University of
Peradeniya, Sri Lanka has been participating in studies on
antimicrobial resistance and residues of animal originated
food, through the World Health Organization (WHO)
program of External Quality assurance System (EQAS)
since 2005. However, these program activities are able to
produce a glimpse of what is the real situation of
ant imi cro bia l use , ant imi cro bia l res ist anc e, and
antimicrobial and other chemical residues in water, plants
and animal originated food in Sri Lanka. The WHO has
als o est abl ish ed the Foo dbo rne Dis eas e Bur den
Epidemiology Reference Group (FERG) in 2010 in which
Sri Lanka collaborated (Havelaar et al., 2015).
The annual health bulletin of Sri Lanka in 2014 reported
the leading 11th cause of hospitalization was intestinal
infectious diseases (128,733 cases) and 9th cause among
the leading cause of hospital deaths. In year 2014 the
ministry allocation for health was 162 billion rupees. This
is mainly spent on recurrent and capital expenditure. The
amount spent on recurrent expenditure was 77 % (Sri
Lanka National Health account 2005-2009). The main
part is spent on curative health which is for the inpatient
and outpatient care. The percentage spent on inpatient/
hospitalized patients was 95% from the recurrent
expenditure (Public hospital governance in Sri Lanka,
2015). From this the percentage spent on gastrointestinal
patient (total number of GIT infectious disease people/
total number of hospitalized patients) was around 2%.
This accounts for 2.37 billion rupees.
In Sri Lanka, Central Food Control administration
comes under the purview of the Ministry of Health. The
Food Act no 26 of 1980 enables the legal authority
regarding the manufacturing, storage, sale and
distribution of food in the country. This is antecedent and
how well it addresses the existing need of the country is of
concern. When recent past is considered there is an
increasing trend towards food borne illnesses and the
public interest stirrup occasionally with frequent news
bulletins. The passive surveillance research already
conducted in Sri Lanka has not yet been adequate to fulfill
the requirement of preventing the economic loss that had
been occurring in the country. Therefore more scope
remains for the research activities in the area of food safety.
There are numerous successful stories where food borne
illness has been successfully controlled by strengthening
the surveillance system in both European and Asian
countries such as Denmark, Thailand and Malaysia
(INFOSAN 2014/ 2015;, Pulse Net 2016).
RISK ANALYSIS AND THE NEED OF FOOD
SAFETY MONITORING AND SURVEILLANCE
SYSTEM IN SRI LANKA
Th e wo rl d- wi de pr ac ti ce d ap pr oa ch fo r th e
identification of food borne illness is the systematic risk
assessment at every step involved to rule out or rule in the
suspected cause. The risk is defined as the probability of
occurrence of an undesired event and its consequences.
The food borne pathogen can be chemical, biological and
physical in nature (Codex Alimentarius, 2010).
Sri Lanka is in need of an active surveillance system to
reduce the disease burden. The country can learn from
these examples and adopt according to its requirements.
The world practiced approach for monitoring food safety
through systematic surveillance at each step along the
food chain is briefly explained.
Once a suspected case of food poisoning is reported
there may be different culprits such as the environment or
the food handlers but most of the time it is directed
towards the food itself. In the ideal situation the samples
will be examined from all the suspected routes of entry.
This will be conducted via microbiological testing
whether to rule in or rule out each cause. When the food
hygiene is considered, it involves sequential steps from
farm to table. The procedure can be categorized as preprocessing stages and post- processing stages.
If meat production is considered the preprocessing stage
involves the sequential steps starting from the farm
environment, animal feed, animals on farm, transport to
the market or lairage, from there to abattoir and processing.
The post processing stage involves storage and retail,
domestic trade and retail catering to the consumers. All
along the chain there are numerous routes from where the
introduction and the transmission of pathogens could take
place. If the chemical agents are considered it can be heavy
metals, pesticides, veterinary drugs and growth promoters,
preservatives and additives. The biological agents are
bacteria, virus and parasites that are from water, soil and
animal cross contamination.
The OIE risk definition is mainly concerned with the
international product import and export. The risk is
defined as “The likelihood of the occurrence and the
magnitude of the consequences of an adverse event to
animal or human health in the importing country during a
specified time period”. Here we are concerned with the
local consumption and this should be identified as the
likelihood of occurrence and the magnitude of
consequences of adverse event to animal/ human health
during the specified time period. According to OIE, Risk
assessment is “A systematic way of gathering, evaluating,
and recording information leading to recommendations
for a position or action in response to an identified hazard
(hazard=food borne illness)”.
According to the Codex alimentarius, risk assessment is
“the identification of biological, chemical and physical
agents capable of causing adverse health effects and which
may be present in a particular food or group of foods”.
33
Food safety monitoring
The risk assessment consists of 4 steps: (i) release
ass ess men t, (ii ) exp osu re ass ess men t, and (ii i)
consequence assessment and (iv) risk estimation.
With regard to food of animal origin at the release
assessment step the veterinarian plays the key role.
Release assessment is the releasing of pathogenic
organism to the environment from the source. This step
identifies the point where the pathogenic agent had
entered the food chain both at pre-processing and the postprocessing. Pre processing stages are in the farm where
apart from pathogenic organisms, the drug residues,
fungal toxins and heavy metals can enter the food chain.
The releasing factors can be categorized as biological,
country and commodity factors. The post processing
stage involves first meat inspection.
Exposure assessment expresses the biological pathways
necessary for exposure of humans to the hazards
(biological or chemical) released from a given risk source
i.e. the frequency of contamination of a food by an agent
and its level in the food over time before consumption. It
should be considered although the pathogenic organism
can be present at one step, at another step it can be
completely destroyed by cooking at a higher temperature
or freezing at a lower temperature.
Consequence assessment describes the adverse health
consequences that may arise as a result of being exposed to
such biological or chemical agent. As indicated earlier,
causing nearly an estimated amount of Rs 2.37 billion loss
and 128,733 cases per year, the foodborne illnesses trigger
considerable losses to the economy and the health of the
people of the country. Apart from the direct financial loss
there is the associated socio-economic loss. The socioeconomic impact of the diseases relates to the loss of
manpower to the government and/or the private sector,
and to the family of the person as a result from being sick.
This is calculated by the disability adjusted life year
(DALY) which is based on the loss of number of years due
to the disease burden (ill health and death).
Based on the results of the risk assessment the risk
estimation is done qualitatively or quantitatively.
Qualitative risk assessment is carried out by developing
the risk matrix where the ultimate risk is categorized as
high, medium and low. Quantitative risk assessment is
carried out by deterministic or stochastic models.
Once the risk is identified it should be managed to an
acceptable level. This is instigated by awareness, policy
making and creating standards. At international level for
importation of food products it is essential to achieve Food
Safety Objectives (FSO) and follow Good Hygiene
Practice (GHP) /Good Manufacturing Practice (GMP).
Considering the local market, the created standard should
be able to provide appropriate level of consumer
protection. Special attention should be paid at the local
market since Sri Lanka is becoming a popular tourist
destination.
In USA, CDC has announced that by reducing the food
related illness by 10%, the Americans have reduced 5
million people getting sick every year. From the consumer
aspect they should be encouraged to buy the products that
are certified and that can be traced back.
In conclusion the need remains for implementing a
proper food safety surveillance system in the country with
a multi-disciplinary approach at every step involved. With
the present economic growth rate of Sri Lanka of 6.2%, the
purchasing ability of general public is improving and the
need for quality food products will be in demand. This will
create a suitable environment for such organized systems.
The government should be made aware of the role of
veterinarian in this context. Further, the research capacity
should be strengthened as to address how the system can
be improved to meet the gap between the public health
challenges and preventing the economic losses to the
country.
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S.L.Vet.J. 2016, 63:2(B): 35-37
HUMAN-MONKEY (Macaca sinica) CONFLICT IN SRI LANKA
SUMMARY: Human- monkey conflict is often reported
due to deforestation and increasing human population
with their needs. This study summarises the findings of
most of the work done in this regard from the year 2000 to
2016. Largely individual monkeys and/or individual
lonely mothers with young ones and secondly the troupes
of monkeys cause conflicts within Kandy municipal area.
Red faced monkeys prefer human food, home garden
produce and insects found in and around houses. In
addition to being a menace, these monkeys, carry some
potentially zoonotic enteric pathogens. The methods
launched to control this conflict were tested such as
capturing, sterilizing and translocating or rehabilitating
and use of repelling devices to change monkeys' trails. The
castration, and ovario hysterectomy on a portion of the
troop in reducing their numbers needs debate. A
completely monkey proof garbage disposal system is also
a must.
INTRODUCTION
With increasing deforestation, human - monkey conflict
in Sri Lanka is on the increase. The increasing human
population growth and exponential increase in their needs
also have contributed to the same (Nahallage, et al., 2008).
In the year 2000, after several written requests reaching
the Veterinary Teaching Hospital (VTH), University of
Peradeniya (UOP), several programs were launched in
this regard. The red faced monkeys (toque macaques/
Macaca sinica) is the culprit animal, making trouble to
daily life of several groups of individuals, shops, schools
and places of worship. This documentation summarises
all efforts of VTH from 2000 to 2016 with regard to human
monkey conflict.
EXTENT OF THE CONFLICT
The Veterinary Teaching Hospital (VTH) launched a
project through Postgraduate Institute of Science (PGIS)
in the year 2000 to study the extent of this problem within
Kandy Municipal limits. It showed that there were two
types of monkeys involved. Firstly, individual monkeys or
individual mothers with kids. Secondly, the troupes.
These monkeys had been possibly tamed and later
released or had been used in circuses to perform various
acrobatic actions by people to make a living (Wijesinghe,
et al., 2005).
These monkeys keep repeatedly coming in search of
tastier human food which can be found easily and, they
stay around and increase their rate of breeding. It is
obvious that the monkey threat has got worse during the
past 15 years and that they dislike arecanut trees and
certain types of banana trees while they eat or destroy all
other home garden produce (Jayalath, 2011). Monkeys eat
not only plant material, but also insects that are found in
and around human dwellings such as those under the roofs.
This is probably why monkeys live in the periphery of the
jungles close to humans. Though, it appears that largely
red faced monkeys (Macaca sinica) are reported to be the
problem, the behavior of other species of monkeys could
also be disturbing (Binduhewa et al., 2005).
Suddenly within Kandy city limits in 2012, several
monkeys were found dead and some of them were found
ill. Such ill and dead monkeys were brought and treated at
the VTH with the help of the Police in Kandy. The
postmortems performed on the dead indicated that they
had possibly consumed a poison. Though this purposeful
poisoning was reported in daily newspapers, nobody
bothered to examine as to why it happened. The monkeys
at Udawattakele, a wildlife protected area in Kandy, in
2014 were not well and feeling ill which was possibly a
viral condition and a few of them even died. This appears
to be an annual occurrence which settles on its own.
COPROLOGIC SURVEY AMONG RED FACED
MONKEYS-ENTERIC PATHOGENS WITH
ZOONOTIC POTENTIAL
In 2015, a coprologic survey (Mendis, 2016) on enteric
parasites, selected bacteria and viruses with zoonotic
potential in Macaca sinica using 105 samples from Kandy
area was performed. In that study, 4 types of protozoan
cysts: Entamoeba coli, Entamoeba histolytica/ dispar,
Giardia spp. and Balantidium coli, 4 types of helminth
eggs: strongyle type/ Strongyloides spp., Trichuris spp.,
Enterobius spp., Bertiella spp. and one bacterial species:
Shigella sp. were found. Some parasites were found more
often from monkeys from areas with close contact with
humans.
CONTROL MEASURES FOR THE CONFLICT
In 2005, all trouble making individual monkeys within
municipal limits in Kandy were caught using different
methods; trapped and/or sedated (Wijesinghe et al., 2005).
These monkeys, after capturing, were surgically castrated
or ovario- hysterectomised, vaccinated against rabies and
tetanus and introduced into a special cage complex. This
cage complex had 8 individual cages into which the
monkeys were introduced, and a common corridor. The
monkeys caught as above, lived in this cage complex for 6
months. One monkey at a time, was released into the
BCG Mendis1 and Ashoka Dangolla 2
Division of Bio resources, Research Center for Zoonosis Control, Hokkaido University, Japan; 2 Department of
Veterinary Clinical Science, Faculty of Veterinary Medicine and Animal Sciences, University of Peradeniya, Sri Lanka
http://doi.org/10.4038/slvj.v63i2.15
1
36
Mendis & Dangolla
common corridor in the cage complex, made contacts with
others and established hierarchy. At the end of the period,
they were all in one troop together, established the
hier arch y and henc e were rele ased to the wild
(Rupasinghe, 2006). This troop however, was not
subsequently monitored.
The medical castration of male monkeys, by injecting
formalin into testicles and using human intra uterine loops
on female monkeys have also been attempted with some
success though the work was inconclusive (Samal et al.,
2015). Is sues on ethics and pain relief were some
problems in that work. However, the impact of performing
such techniques only on a proportion of males or/and
females in a troop, on reducing their numbers is
questionable.
A village in Hali-ela area launched a program to control
human-monkey conflict after holding, several meetings
with the villagers. All these attempts were not to kill,
destroy or to develop hatred towards monkeys but to
educate humans to live with the monkeys in harmony. It
was pointed out in such meetings that the most feasible
among the few temporary solutions was to, capture,
surgically sterilize, vaccinate and to release either to the
same location or to translocate. The carpenters from the
village built community monkey traps. One person was
employed to operate the trap (Wijesinghe et al., 2009).
The people in the area were also made to understand that
the welfare and the genetic make-up of these monkeys
must be looked into. This program with the assistance of
the Department of Wildlife Conservation subsequently
became popular in other areas of the country.
Mahakanda village in Kandy conducted such a capture
programme and a substantial proportions of animals from
several troupes were caught, surgically operated and were
translocated (Jayalath, 2011). However, the monkeys
from the surrounding areas infiltrated in about 6-7 months.
These new monkeys though initially not destructive,
became gradually so (Jayalath and Dangolla, 2011).
In 2014, the University of Peradeniya, imported 5
monkey repelling electrically operated electronic devices
from India. These were fitted on pre-determined strategic
points within the university premises after a detailed study
of troops and their trails in the area. The monkeys however
did not change their trails or would not be disturbed by this
equipment. In addition, these devices were producing
noises that were disturbingly audible for people.
Applying luminous paint on monkeys to make other
monkeys scared, making the monkeys eat hot chilies and
clipping their hair were studied as remedial measures all of
which the responses were temporary and transient. During
2012, it was brought to our knowledge that in Ukuwela,
Matale, a certain agricultural land was never attacked by
monkeys while all other lands around were inhabited and
destroyed by them. The reason indicated was that the
construction of the fence of this particular land had been
done on an auspicious time according to the fortune of the
owner of the land. There is also a belief that hanging a
fresh goat skin on the monkey trail would repel monkeys.
It is clear that a complete monkey proof garbage
disposal method must be introduced to reduce this
problem. A special monkey proof waste bin has been
introduced in Polonnaruwa area as a solution to the
conflict. Most inhabitants do not take an extra effort to use
the garbage bins properly and throw garbage from a
distance into the bin. To use a monkey proof waste bin,
people must take an effort to dispose their own waste.
EDUCATIONAL PROGRAMMES AMONG
SCHOOL CHILDREN
Several schools sent their children to study on humanmonkey conflict resolution to VTH. They were used to
make observations in these experiments and were taught
that a real life problem needs pooling knowledge from
various disciplines; i.e. a multi-disciplinary approach. In
addition, this project was also used to examine the use of
“hypothesis testing” lesson in the GCE A/L school
syllabus in Science curriculum at that time (Rupasinghe
and Dangolla, 2005).
An initial island-wide survey with several educational
programs was conducted by VTH. The children in the
GCE advanced levels Science stream were also used in
these programs (Rupasinghe, 2006). These educational
prog rams and subs eque nt inve stig atio ns incl uded
teachings on potential zoonotic infections that could be
transmitted to humans from monkeys. The school children
learn these principals quicker than expected (Wijesinghe
et al., 2003).
CONCLUSION
A survey on the international literature on the monkey
human conflict, show that there is no permanent solution
for this problem. Proper and responsible garbage disposal
seems to be a must. In agricultural areas in Sri Lanka this
recommendation may not work. None of the different
approaches adopted in India, Thailand or Malaysia have
shown total effectivity. Resolving this problem in cities
and agricultural areas, while appreciating the living rights,
welfare and genetic issues of both humans and monkeys is
urgently needed. It is timely that the Kandy municipality
takes lead attempting to solve this problem, examine the
potential benefits and losses of the proposed solutions and
make decision on mass scale introduction and adoption of
a remedy to ensure decent living standard for people
within municipal limits.
REFERENCES
Binduhewa, C.H.J., Silva, I., Dangolla, A. (2005).
Comparative study on the behavior patterns of the
captive and free ranging monkeys of Sri Lanka. Sri
Lanka Veterinary Journal. 52 (1&2B), 34-35.
Jayalath, P.P. (2011). Evaluation of the impact of capture
and translocation of trouble making toque moneky
(Macaca sinica) troops in Mahakanda village, as a tool
of mitigating the Human-macaque conflict. MSc thesis,
Postgraduate Institute of Science, University of
Peradeniya.
Human monkey conflict
Jayala th, P.P., Dang olla, A. (2 011). Cap ture an d
translocation of trouble making torque monke ys
(Macaca sinica) in Mahakanda: Lessons learnt.
Proceedings of the Annual Scientific sessions of the Sri
Lanka Veterinary Association, Institute of Continuing
Education in Animal Production and Health,
Gannoruwa, 35.
Mendis, B.C.G. (2016). Coprologic survey in monkey
troops (Maccaca sinica) in three selected sites in Kandy.
MSc Thesis, Postgraduate Institute of Science,
University of Peradeniya, Sri Lanka
Nahallage, C. A.D., Huffman, M .A., Kuruppu, N.,
Weerasingha, T. (2008). Diurnal primates in Sri Lanka
and peoples' perception of them. Primate Conservation,
23, 81 - 88. https://doi.org/10.1896/052.023.0109
Rupashinge, M. (2006). Introducing Scientific method for
GCE (Advanced level) student projects using monkey
menace as an example. MSc thesis, Postgraduate
Institute of Science, University of Peradeniya
Rupasinghe, M., Dangolla, A. (2005). Introducing the
Scientific method through GCE (A/L) student projects.
Proce edings of Perad eniya Unive rsity Resear ch
Sessions, Vol 10, Appendix II, vii
Samal, K.S., Kumara, Y.H.P.S.N., Nishanantha, K.,
Jayasinghe, M.G.C.M., Ariyarathne, H.S., Amarasinghe,
37
A.A.A.W.K., Dangolla, A. and Alexander, P.A.B.D.
(2014). A preliminary study to evaluate selected
contraceptive methods in controlling reproduction in
toque monkey (Macaca sinica). Proceedings of Annual
Scientific Sessions of Sri Lanka Veterinary Association.
Oak Ray Hotel, Kandy,71.
Wijesinghe, S.J., Dangolla, A., Rupasinghe, M.,
Binduhewa, C.J.H., Silva, I.D. and Dikkumbura, A.W.
(2005). A possible answer to the emerging monkey
menace i n Kandy. Pro ceeding s of Pera deniya
University Research Sessions, Vol 10, Appendix I, vii
Wijesinghe, S.J., Dangolla, A., Rupasinghe, M., Perera,
G.I.S. (2003). A study on short term memory of school
children on zoonotic diseases. Proceedings of
Peradeniya University Research Sessions, Vol 10,
Appendix III, ix
Wijesinghe, S.J., Jayasinghe, M.G.C.M., Kularathne, M.N.,
Dangolla, A., Nissanka, A.H.S., Vijitha Nanda Kumara,
G.W. (2009). Pilot project on solving “Monkey
(Maacaca sinica) Menace in Hali-Ela Area. Abstracts of
the National Symposium on Promoting knowledge
Transfer to Strengthen Disaster Risk Reduction and
Climate Change Adaptation, 7-8th July 2009, BMICH
Colombo ISBN 978-955-9321-11-8, 57
S.L.Vet.J. 2016, 63:2(B): 38
Letter to the Editor
Cattle rabies in Nawagattegama Veterinary range
Dear Sir,
Rabies is an endemic disease in Sri Lanka. Most often,
the disease is reported in domestic or free roaming dogs. It
is rapidly progressive meningioencephalitis caused by the
Rabies virus or other lyssa viruses. There are a
considerable number of human deaths due to Rabies in Sri
Lanka and most of these human rabies cases are linked to
dog bites. Therefore, there is an enormous attention to
prevent and control rabies in the dog population of Sri
Lanka. However, it is important to note that the rabies in
livestock can also be a source for human rabies. Due to
lack of awareness of rabies in cattle, goat and sheep, exact
number of cases remains undiagnosed, hence, under
estimated. Here we reported five cases rabies in cattle in
Nawagattegama veterinary range. Three of these cases
were reported in the same farm located in Konkadawala
Grama Niladari division.
One year old Jersey cattle was reported to show
aggressive behaviour, drooling of saliva, bellowing and
was completely off food. Clinical examination revealed
that the animal has hyperthermia and nystagmus. The
affected cattle was in a herd of six animals and was reared
under semi intensive management system. As the
condition was suspected of rabies, it was advised to keep
the animal in isolation under observation. Two days later
the affected animal died.
Another animal, in the same herd of the previous case,
started of showing similar clinical signs and died two days
later.
On 04th of April 2016, a two month old female calf in the
Kahandawa GS division also died after developing
clinical signs described above. Head of the dead animal
was packed in ice and send to Medical Research Institute,
Colombo to check for rabies virus. Laboratory results
confirmed that the animal was positive for rabies.
A young cattle, aged one year and two months also
developed anorexia and aggressive behaviour on 16th
April 2016 and died on the following day. Head of this
animal was also send to MRI to check for rabies and was
confirmed rabies positive.
On 14th of May 2016, two year old cattle developed
similar sign and died on the same day.
According to the farmers who owned these animals,
they were not bitten by dogs. However, they were tied in
nearby paddy fields during night time. It is unclear
whether these cattle infected by rabies virus through a
rabid dog, wild animals or through bats. Due to public
health risk as well as to the economic losses to the rural
farmers, it would be important to vaccinate livestock
animals against rabies.
H.A.S.I. Harischandra
Government Veterinary Surgeon, Department of Animal
Production and Health, Veterinary Office,
Nawagatthegama, Sri Lanka
http://doi.org/10.4038/slvj.v63i2.17
S.L.Vet.J. 2016, 63:2(B): 38
Letter to the Editor
Can GnRH vaccine be used on captive elephants to
postpone musth?
Dear Sir,
Four client owned, captive adult male elephants were
given injections of a GnRH (gonadotropin releasing
hormone vaccine; 800µg per dose). The hormone
administrations were planned so that the last injection was
administered approximately one month before the
anticipated date of musth. Two semi wild male elephants
from the Pinnawela elephant orphanage also received the
vaccine. Venous blood was collected immediately before
the vaccine injection for assay of testosterone.
Of the client owned elephants, one received all 3 doses
and demonstrated a 3-month delay in the onset of musth
together with a marked reduction in aggressiveness. Two
elephants that received 2 injections showed signs of musth
around the expected time, however, there was a reduction
in the aggression. Conversely, one animal that received
one injection came into musth, two months prior to the
expected date and had a prolonged period of musth.
In the semi wild elephants, one had apparently been in
continuous musth for almost 2 years when the first
injection was administered and this animal came out of
musth immediately after the injection and had minimal
aggression. However, musth reappeared approximately 5
months after the second dose. The other semi wild
elephant had a history of irregular musth twice a year but
responded well to the vaccine by demonstrating delayed
commencement of musth by approximately 8 months
after the second injection. Testosterone levels were
reduced in all four client owned elephants and a semi wild
animal while the other semi wild elephant showed no
changes in the testosterone levels.
These preliminary findings indicate a postponement of
musth, albeit for short periods following GnRH vaccine in
captive and semi wild elephants. It also appears that
musth-related aggression in the male elephants is complex
and possibly multifactorial and is not solely reliant on
testosterone. Further studies with a large number of
animals to include captive, semi wild and perhaps wild
elephants will help to confirm the findings reported
herein. Client compliance was a major impediment in the
conduct of this study.
TMSK Piyadasa, A Dangolla, C Rajapakshe,
E Rajapakshe, N Horadagoda, PGA Pushpakumara and
P Windsor
Departments of Veterinary Clinical Science, Faculty of
Veterinary Medicine and Animal Science, University of
Peradeniya, Peradeniya, Sri Lanka
http://doi.org/10.4038/slvj.v63i2.16