Abstract:
A cross-sectional epidemiological survey was conducted to check the seroprevalence of 3
reportable equine diseases viz., equine infectious anemia (EIA), glanders, and piroplasmosis
in 5 draught equine populated urban areas (Lahore, Gujranwala, Faisalabad, Multan and
Bahawalpur) of Punjab. Assuming the expected prevalence to be 50 percent (unknown status)
with confidence limits of 95% and a desired absolute precision of 5%, a total of 430 blood
and sera (comprising 332 donkeys, 65 horses and 33 mules) along with relevant information
were randomly collected. The samples were subjected to microscopic examination (thin and
thick Giemsa stained smears), hematological analysis and commercial enzyme linked
immunosorbant assay (ELISA). Sera were tested for EIA through ELISA (VMRD, Inc.,
USA), for glanders through rose Bengal plate agglutination test (RBT) and a commercial
competitive ELISA (cELISA, VMRD, Inc., USA) was used for piroplasmosis screening.
Positive and negative samples were identified on the basis of degree of agglutination (++ to
++++ considered positive) in case of RBT and as per instructions of the manufacturer
regarding ELISAs. Data thus generated was analyzed by using epidemiological software tools
(EpiinfoTM, WINPEPI, Survey Toolbox and IBM SPSS). Results: Giemsa stained smears
were negative for blood parasite. None of the samples was positive for EIA, indicating
towards possible disease free status or failure of the disease to mount immune response in
sampled equine population. Seroprevalence of glanders was found to be 7.9% (n=34) with
highest prevalence in Lahore (11.6%) followed by Bahawalpur (10.3%), Multan (7.3%),
Faisalabad (6.7%) and Gujranwala (5.3%). Significantly different (P<0.05) seroprevalence
was recorded in horses (16.9%) than mules (12.1%) and donkeys (5.7%). Seroprevalence
recorded in young (< 5 years), adult (< 10 years) and old (> 10 years) equines was 7.9%,
9.2% and 5.2% respectively. Prevalence was significantly different (P<0.05) in males 5.7%
and female 12.59% equine. Multivariable analysis indicated that female equines (OR = 2.3,
95% CI= 1.16–4.77) and and those sharing water sources or access to communal water
troughs (OR =2.7, 95% CI=0.93-7.86) were more likely to be sero-positive for glanders.
Seroprevalence of piroplasmosis was found to be 52.5% (T. equi=41.2% and B.
caballi=21.6%; P<0.05) in the selected areas. Forty four (10.2%) sera were found positive for
both piroplasms (mixed infection). Significantly higher (P<0.05) seroprevalence was
recorded in Lahore (95.6%, n=66) than Faisalabad (64%, n=96) than Multan (52.9%, n=36),
Gujranwala (50.7%, n=38) and Bahawalpur (50%, n=34). Seroprevalence of piroplasmosis
was 69.2% in horses (T. equi=56.9%, n=37; B. caballi=24.6%, n=16) followed by 63.6% in
mules (T. equi 48.5%, n=16; B. caballi 30.3%, n=10) and 48.2% in donkeys (37.3%, n=124).
Adjusted analysis for possible confounding factors revealed that equines living alone or with
equine cohorts (OR=1.30, 95%CI=0.84–2.01), with ticks infested equine cohorts (OR=1.19,
95%CI=0.65-2.18) whose owners did not practice tick control (OR=1.43, 95%CI=0.93-2.21)
were more likely to be sero-positive for piroplasmosis. Hematological alterations observed in
RBT (glanders) positive equines regarding white blood cell counts, red blood cell counts,
hemoglobin concentrations and hematocrit in all there RBT positive equine species.
Macrocytic hyperchromic type of anemia was observed in all RBT positive horses, mules and
donkeys. Hematological analysis of piroplasmosis positive equines indicated decreased white
blood cell counts, red blood cell counts PCV and hemoglobin concentration values in
seropositive horses, donkeys and mules. Erythrocytic indices pointed towards microcytic
hyperchromic type of anemia in piroplasmosis sero-positive equines. In conclusion, the high
RBT based prevalence of equine glanders indicates towards possible endemic nature of this
disease in Pakistan. Potential role of communal water sources and faulty management in the
spread of equine glanders requires further investigations through carefully designed
experimental studies. Seroprevalence of piroplasmosis was alarmingly high in the selected
population which could be attributed to the faulty housing, lack of tick control practiced by
owners and lacuna regarding knowledge among animal health professionals about the correct
diagnosis and treatment of T. equi and B. caballi.