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Seroprevalence of PPR among Sheep and Goats of Different Agroclimatic Zones of Odisha

A. Hota S. Biswal N. Sahoo M. Rout D. Chaudhary A. B. Pandey D. Muthuchelvan
Vol 8(4), 296-302
DOI- http://dx.doi.org/10.5455/ijlr.20171028023420

The objective of this study was to demonstrate the seroprevalence of peste des petits ruminants (PPR) among small ruminant population of Odisha. The disease is endemic in the state and in India causing huge economic loss to the farming community. A total of 506 serum samples (from 217 sheep and 289 goats) from unvaccinated flocks were collected between October-2015 and April-2016 across 10 agroclimatic zones of Odisha through random sampling and subjected to competitive ELISA to detect antibodies against PPR virus (PPRV). The result indicates an overall seroprevalence rate of 48.42%, with 44.7% contributed by sheep (45.74% male and 43.90% female) and 51.21% by goats (48.15% males and 53.89% females). The agroclimatic zone wise prevalence rate among sheep varied between 100% and 0%, while among goats varied between 69.44% and 29.63%. The study indicates a high prevalence rate of PPR in the state of Odisha and warrants regular vaccination of small ruminants to develop better immunity to prevent the infection.


Keywords : Seroprevalence PPR c-ELISA Sheep Goat Odisha

Introduction

The World Organization for Animal Health (WOAH-OIE) defines Peste des petits ruminants (PPR) as an acute, highly contagious, notifiable and economically important transboundary viral disease affecting small ruminants. It is also called as ‘Goat Plague’ and clinically resembles with Rinder Pest (RP) in cattle, the later being characterized by pyrexia, conjunctivitis, oculo-nasal discharges, necrotizing and erosive stomatitis, diarrhea and bronchopneumonia followed by either death or recovery (Gomes et al., 2016). According to the FAO estimates, an average economic loss of $2,972.5 million/year likely to occur pertaining to morbidity, mortality, production losses and treatment cost of PPR, during 2012-2017 in SAARC region among which, India alone contributed $2569.00 million/year (Kumar et al., 2014). In India, PPR was first reported with an outbreak among sheep with 25% mortality in Arasur village, Villipuram district of Tamil Nadu in March 1987, where the characteristic clinical signs pertaining to PPR were confirmed (Shaila et al., 1989). Thereafter, reports of PPR outbreak across many states of India were made available and India became endemic to this viral infection with due course of time (Balamurugan et al., 2014a; Singh et al., 2004) following the latest report from north eastern states (Meghalaya, Assam, Manipur, Nagaland, Arunachal Pradesh, Tripura and Mizoram) of India with a prevalence rate of 11.63% (Balamurugan et al., 2014b). Odisha being situated in the eastern coast provoked us to undertake the research work to estimate the prevalence status of the disease in the state. Competitive ELISA (c-ELISA), agar gel precipitation test (AGPT) and virus neutralization tests were employed to detect antibodies against PPR virus (PPRV) (Santhamani et al., 2016). The c-ELISA assay based on the inhibition of binding of monoclonal antibody (MAb) to antigen in the presence of PPRV antibodies in test sample is considered highly sensitive and specific, thus employed in the present study to record seroprevalence of PPR among sheep and goat population of Odisha.

Materials and Methods

Study Design

Odisha is located in the eastern coastal part of India, surrounded by the states of West Bengal, Jharkhand, Chhattisgarh and Andhra Pradesh. By area, it is the 9th largest state of India consisting of 10 agroclimatic zones and 30 districts. Random sampling was carried out from unvaccinated sheep and goat flocks of the state during October 2015 to April 2016. A total of 506 serum samples were collected comprising of 217 sheep (from 25 districts) and 289 goats (from 29 districts) of either sex and irrespective of age (Table 1 and 2). All the samples were collected from non-descriptive breeds of sheep and goats from local accessible areas of the concerned districts of the state. The sample size from each area varied due to constraints in accessibility to localities, co-operation of farmers and number of animals available in the area. The samples collected were stored at -20oC and then carried to IVRI, Mukteswar for PPRV antibody detection.

 

Table 1: Overall species wise and sex wise seroprevalence of PPRV infection

Sheep Goat
Male Female Total Male Female Total
Total serum Taken 94 123 217 135 154 289
Positive (%) 43 (45.74) 54 (43.9) 97 (44.7) 65 (48.15) 83 (53.89) 148 (51.21)
χ2 0.073 0.951
P- Value 0.787 0.3294
χ2 2.104
P- Value 0.1469

Table 2: Seroprevalence of PPR with respect to different agroclimatic zones and districts of Odisha

Sl. No. District Agro-climatic zones of Odisha Total positive/Total sample tested (%)
Sheep Goat Sheep Goat
1 Sundergarh North Western Plateau 1/7(14.29) 3/10(30) 5/16(31.25) 9/19(47.37)
2 Deogarh 4/9(44.44) 6/9(66.67)
3 Keonjhar North Central Plateau 10/21(47.62) 10/15(66.67) 25/36(69.44)
4 Mayurbhanj 10/15(66.67) 15/15(100)
5 Bhadrak North Eastern Coastal Plain 9/10(90) 8/11(72.73) 24/39(61.54) 8/27(29.63)
6 Baleswar 0/14(0) 0/16(0)
7 Jajpur 15/15(100)
8 Puri East & South Eastern Coastal Plain 1/11(9.09) 0/10(0) 11/54(20.37) 20/50(40)
9 Cuttack 0/9(0) 6/10(60)
10 Khordha 4/7(57.14) 4/7(57.14)
11 Nayagarh 3/8(37.5) 5/9(55.56)
12 Jagatsinghpur 3/10(30) 3/7(42.86)
13 Kendrapada 0/9(0) 2/7(28.57)
14 Rayagarh North Eastern Ghat 1/8(12.5) 6/11(54.54) 4/11(36.36) 25/38(65.79)
15 Kandhamal 10/10(100)
16 Ganjam 3/3(100) 1/7(14.29)
17 Gajapati 8/10(80)
18 Koraput Eastern Ghat High Land 9/9(100) 5/11(45.45) 19/19(100) 14/21(66.67)
19 Nabarangapur 10/10(100) 9/10(90)
20 Malkangiri South Eastern Ghat 0/9(0) 4/8(50) 0/9(0) 4/8(50)
21 Kalahandi Western Undulating Zone 9/9(100) 6/11(54.54) 12/17(70.59) 9/23(39.13)
22 Nuapara 3/8(37.5) 3/12(25)
23 Bargarh Western Central Table Land 4/11(36.36) 4/21(19.05) 27/48(56.25)
24 Sambalpur 1/5(20) 3/5(60)
25 Bolangir 10/11(90.91)
26 Sonepur 2/5(40) 3/5(60)
27 Boudh 1/3(33.33) 6/8(75)
28 Jharsuguda 0/8(0) 1/8(12.5)
29 Anugul Mid Central Table Land 5/8(62.5) 3/9(33.33) 8/16(50) 7/19(36.84)
30 Dhenkanal 3/8(37.5) 4/10(40)
χ2 62.974 21.097
P- Value <0.0001 0.0122

Screening of Sera Samples by c-ELISA

The c-ELISA indigenously developed by the National Morbillivirus Referral Laboratory, Division of Virology, IVRI, Mukteswar, which is based on the principle of inhibition of binding of monoclonal antibody to PPRV antigen in the presence of PPRV antibody present in field serum. This results in reduced color development when anti-mouse antibody conjugated to HRPO (conjugate) is used for tracing the binding of monoclonal antibody.

The ELISA plates were coated with PPRV antigen (50 ml/ well) at 1:100 dilutions in PBS and then plates were incubated at 37°C for one hour under constant orbital shaking. Unbound antigens were washed off the using diluted PBS (1:4) three times. Then 40 ml of blocking buffer (PBS having 0.3 % negative serum and 0.1% Tween 20) was added in all the wells of the plates. Additional blocking buffer were added in Conjugate control (Cc) wells and MAb control (Mc) wells at the rate of 60ml and 20ml, respectively. Then 20 ml of the serum samples were added to each well of the plate in duplicate sets. Then 20ml of the control serum (negative, weak positive and strong positive) were added in respective wells. This was followed by addition of 40ml of monoclonal antibody (1:100 dilutions in blocking buffer) in all the wells of the plate except Cc wells. After incubation of the plates was done for one hour under constant orbital shaking, following washing the wells using diluted PBS (1:4) three times. Rabbit anti-mouse-HRPO conjugate (diluted 1:1000 in blocking buffer) was added to each well of the plate @ of 50ml/well. Further incubation of the plates was done for one hour at 37°C under constant shaking. Then the wells were washed using diluted PBS (1:4) three times. Substrate-chromogen mixture (OPD containing H2O2) were added in each well (50ml/well) along with a blanking module and color reaction is developed for 10 min followed by stopping with equal volume (50ml/well) of 1M H2SO4. Finally Optical Density (OD) of the wells was measured at 492nm after blanking the wells of the plate using EDI software (developed by the Food and agriculture Organization /International Atomic Energy Agency Laboratory, Siebersdorf, Austria). Result was denoted in terms of percentage inhibition (PI) values.

PI values for each well can be calculated manually using the following formula:

PI = 100 – {(OD of test sample ¸ OD of Mc) x 100}

Statistical Analysis

The results were considered statistically significant at p<0.05. Chi-square test using RealStats-2007 (accessible at www.real-statistics.com/) was used to determine the difference in susceptibility of animals with respect to sex and agro-climatic zones.

Results and Discussion

Out of 506 sera processed through c-ELISA for detection PPRV antibody, 245 (48.42%) samples were found positive. Out of 217 sheep sera processed, the prevalence of PPRV antibody was found to be 44.7% with 45.74% in male and 43.90% in female. Similarly in goats, out of 289 serum samples tested, the overall seroprevalence rate was 51.21% with 48.15% in males and 53.89% in females (Table 1). The seropositivity for PPRV might be due to its endemicity as reported by Hegde and Deo (2017) to be a major disease of goats in Odisha, Bihar and Uttar Pradesh. As per the AICRP annual report (2016), a total of 11 PPR outbreaks were recorded affecting 337 sheep and goats, causing 129 deaths in different districts of Odisha. Previously Nayak et al. (1997) reported outbreak of PPR in a goat breeding farm Ghatagaon, Keonjhar during November 1995, which is considered to be the first outbreak of the state, with an overall mortality rate of 53.3%. Thereafter, prevalence of 0.75% among sheep and 15.7% among goats was recorded between the year 1998 and 2003 (Singh et al., 2004).The movement of infected animals across the borders might often act as a source of infection to the healthy animals. Overall seroprevalence of PPRV antibody among population of small ruminants in West Bengal, India was found to be 31.23% (Pal et al., 2014). Prevalence of 45.66% in sheep and 38.54% in goats were reported in different villages of 52 districts in five states of India during 2011 (Balamurugan et al., 2014c). The higher incidence in goats (70.37%) than in sheep (65.32%) was found by Saritha et al. (2014) in different agro-climatic zones of Andhra Pradesh. An overall prevalence of 55.55%, with higher incidence in goats (66.66%) than in sheep (35.71%) from Jammu region of Jammu and Kashmir (J&K) was reported by Mahajan et al. (2013).

During the study, both agroclimatic zone and district-wise prevalence were recorded. Out of the total sheep samples processed, the highest prevalence of PPRV antibodies was recorded in Eastern Ghat High Land (100%) zone and the lowest in South Eastern Ghat (0%), whereas out of the total goat sera processed, the highest prevalence was recorded in North Central Plateau (69.44%) zone followed by the lowest in North Eastern Coastal Plain (29.63%) (Table 2). The variation among the different agroclimatic zones with respect to sheep and goat were found to be at significant level (p<0.05). Out of the sheep sera collected from different districts, the highest prevalence was recorded in the districts Jaipur, Ganjam, Koraput, Nabarangapur and Kalahandi (100%) followed by the lowest in the districts Baleswar, Cuttack, Kendrapada, Malkangiri and Jharsuguda (0%). Similarly, out of goat sera processed from different districts, Mayurbhanj and Kandhamal (100%) were recorded with the highest prevalence followed by the lowest in Baleswar and Puri (0%) (Table 2). The results and significance level may vary upon processing of more number of serum samples. The cause of variation in prevalence with respect to different zones and districts may be corroborated with the varying climatic conditions (Chauhan et al., 2012) like temperature, humidity, rainfall and soil type.

Mostly endemicity of PPR in small ruminants pertains to transmission of infection, either through close interaction of healthy animals with the affected ones or due to nomadic grazing and transboundary movement of affected animals. Spread of infection also occurs via infected materials, fomites, contaminated water and feed troughs and bedding etc. (Banyard et al., 2010). The disease causes a high morbidity (100%) and mortality (20-90%) among small ruminants (Singh et al., 2014). The matter of concern is about the dependence of millions of small, marginal, landless farmers upon these animals for their livelihood and daily earnings, for which these animals are considered as poor man’s cow in India. The predisposing factors towards the prevailing infection can be linked with poor nutritional management, environmental stress, concurrent parasitic and/or bacterial infection, ultimately enhancing the severity of the disease (Alemayehu, et al., 2015). Frequent economic losses in India have been claimed by several authors due to this disease (Singh et al., 2014, Singh and Prasad, 2008; Venkataramanan et al., 2005). In order to check further losses, it is necessary to take preventive and control measures in endemic areas. Also trade limitation is another constrain to the country’s economy, calls for a global eradication program which has already been started by OIE and FAO (Singh and Bandyopadhyay, 2015). Implementation of extensive vaccination programme using a thermostable vaccine and mass awareness camps could be the ideal approach to minimize the infection and losses.

Conclusion

The present study indicates a high prevalence rate (48.42%) of PPR in the state of Odisha, which might the first time such an extensive study done covering all the thirty districts of the state and warrants regular effective vaccination of small ruminants to prevent the infection

Acknowledgement

All the farmers across the state of Odisha who cooperated during sample collection are gratefully acknowledged. All the CDVOs, BVOs, VAS, AVAS and paravets of the corresponding areas are heartily thanked for extending their cooperation during the study. Vice Chancellor of O.U.A.T. and Dean, C.V.Sc. and Animal Husbandry are thankfully acknowledged for giving approval of this study, as a part of thesis submitted to the University.

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