Seroprevalence of Contagious Caprine Pleuropneumonia (CCPP) in Rupandehi and Palpa Districts of Nepal

Basanta Kumar Adhikari1*, Deepak Subedi1*, Sumit Jyoti1, Krishna Kaphle2, Chet Narayan Kharel3, Doj Raj Khanal4

1Paklihawa Campus, Institute of Agriculture and Animal Science, Tribhuvan University, Rupandehi, Nepal; 2Department of Theriogenology, Paklihawa Campus, Institute of Agriculture and Animal Science, Tribhuvan University, Rupandehi, Nepal; 3National Avian Disease Investigation Laboratory (NADIL), Bharatpur, Chitwan, Nepal; 4National Animal Health Research Centre, Nepal Agricultural Research Council (NARC), Khumaltar, Lalitpur, Nepal.

Editor | Muhammad Abubakar, National Veterinary Laboratories, Park Road, Islamabad, Pakistan.

Received | December 06, 2021; Accepted | April 11, 2022; Published | May 18, 2022

*Correspondence | Basanta Kumar Adhikari and Deepak Subedi, Paklihawa Campus, Institute of Agriculture and Animal Science, Tribhuvan University, Rupandehi, Nepal; Email: [email protected], [email protected]

Citation | Adhikari, B.K., D. Subedi, S. Jyoti, K. Kaphle, C.N. Kharel and D.R. Khanal. 2022. Seroprevalence of contagious caprine pleuropneumonia (CCPP) in Rupandehi and Palpa Districts of Nepal. Veterinary Sciences: Research and Reviews, 8(1): 23-29.

DOI | https://dx.doi.org/10.17582/journal.vsrr/2022.8.1.23.29

Keywords | c-ELISA, First detection, Goats, Nepal, Risk factors

Copyright: 2022 by the authors. Licensee ResearchersLinks Ltd, England, UK.

This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (https://creativecommons.org/licenses/by/4.0/).

Introduction

Contagious caprine pleuropneumonia (CCPP) is a rapidly spreading, highly infectious, mycoplasmal disease of small ruminant characterized by respiratory distress, high morbidity and mortality (Arif et al., 2007; Iqbal et al., 2019). CCPP in goats and sheep is caused by Mycoplasma capricolum subspecies capripneumoniae commonly known as Mccp and previously known as Mycoplasma biotype F38 (Iqbal et al., 2019). The major source of contamination is direct contact with the infected animals via inhalation of infected aerosols (Thiaucourt et al., 1996). CCPP usually has per-acute, acute and chronic forms. In the per-acute form, the animal dies within 1-3 days with minimal clinical manifestation while signs of high fever (41-43˚C), lethargy, anorexia, coughing and laboured breathing are seen in acute forms of the disease (OIE, 2009). Debility, chronic cough and nasal discharges are the major clinical signs in the chronic form of the disease (OIE, 2009). The pathogenicity of CCPP is usually restricted in the lungs and pleura with characteristic fibrinous pleuropneumonia (Ahaduzzaman, 2020). Consolidation of lungs (100%), pleural adhesion (72.72%) and alveolar exudation (90.9%) are the major gross findings and septal peribronchiolar fibrosis (81.8%), fibrinous pleuritis (63.63%) and peribronchiolar cuffing of mononuclear cells (54.54%) are the typical microscopic findings of the disease (Hussain et al., 2012).

CCPP is an important transboundary disease especially in low and middle-income countries and causes significant trade and economic disturbances in various parts of the world (Ahaduzzaman, 2020; Jores et al., 2020). Nepal has not reported any outbreaks of CCPP to OIE but the clinical descriptions of CCPP has been reported in Nepal (IDRC, 2016). Different studies show the prevalence and incidences of CCPP in India (Iqbal et al., 2019). Seroprevalence of CCCP was 33.67 % in Nagpur. (Ingle et al., 2008), 10.65 % in Jabalpur (Gupta et al., 2016), 9.93% among Himalayan pashmina goats of Ladakh (Parray et al., 2019), 16.05 % in goats and 20.24 % in sheep population of Maharashtra (Suryawanshi et al., 2015) and 8.11% in Uttar Pradesh of India (Jain, 2015). The presence of CCPP in India, Tibet of China (Chu et al., 2011; Yu et al., 2013), and Pakistan (Awan et al., 2010; Wazir et al., 2016) can increase the risk of an outbreak of CCPP in Nepal.

In the fiscal year 2019/20, Nepal imported 327,897 live goats worth Rs 1.93 billion. Porous Nepalese borders with India, weak disease surveillance, and poor quarantine facilities increase the risk of CCPP in Nepal. Widespread movement of livestock between India and Nepal might favor a spread of Mccp in small ruminants of Nepal (Parajuli and Acharya, 2019). The objectives of our study were to evaluate the seroprevalence of Mccp in goats of Rupandehi and Palpa districts of Nepal and identify the risk factors associated with Mccp seropositivity.

Materials and Methods

Study design, area, and population

The cross-sectional study was conducted for 3 months between January 2020 and March 2020 to detect the presence of antibodies against Mycoplasma capricolum capripneumoniae. The study was conducted in the selected Rohini rural municipality of Rupandehi and the Tinau rural municipality of Palpa (Figure 1). Rohini rural municipality has a total area of 64.62 sq. km. It is located in the eastern south part of Rupandehi district at 27˚51’21” N latitude, 83°54’97”E longitude, and at an altitude of 92.1 m. This municipality shares its southern boundary with the Uttar Pradesh state of India. Tinau rural municipality of Palpa district has a total area of 202 sq. km. It is located in the southern part of the district on 27˚68’13” N to 27°46’ 53 “N latitude, 84°40’32”E to 83°30’57 “E longitude and at an altitude between 300 m to 1220 m.

 

The study population was goats of both local and exotic breeds in the study area. Local breeds include Terai in the lower plains of Rohini rural municipality, Rupandehi district, and Khari in mid-hills of Tinau rural municipality, Palpa district. The housing system of the goat includes the intensive and semi-intensive type of housing system.

Sample size calculation

The sample size necessary for the calculation of the prevalence of CCPP was be calculated by EpiTools Epidemiological Calculators by Ausvet (Sergeant, 2018). According to the literature review, the prevalence of CCPP in neighboring state i.e. Uttar Pradesh of India was found to be 10.5 % (Jain et al., 2015). As Uttar Pradesh is very close to our study area, its prevalence was taken as a reference and the desired sample size was calculated with expected precision of 5 % at a confidence level of 95%. The calculated minimum sample size required for the study was 145 based on the total goat population (392,791) of the two districts i.e., Rupandehi (185,332) and Palpa (207,459) as per the livestock statistics of Nepal, 2017 (MOLD, 2017). Only 101 samples were collected due to the limited time frame, COVID-19 crisis and financial constraints. Despite collecting 101 serum samples only 89 samples were included in the study. The remaining serum samples were discarded due to haemolysis. Out of 89 serum samples, 30 were from Rohini rural municipality of Rupandehi district and 59 were from the Tinau rural municipality of Palpa district.

Questionnaire administration, sample collection, and laboratory analysis

A semi-structured questionnaire was administered to collect data from the goat farmers by the purposive sampling method. The close and open-ended questionnaire was pretested to ensure its validity and verbal consent was obtained from the respondent by explaining the objectives of the survey.

No specific criteria were set for sample collection. Five millilitres (5ml) of blood samples were collected from the jugular vein of each properly restrained selected goat with the help of a 5 ml syringe and 21-gauge needle using plain vacutainer tubes. The collected blood samples were labelled and kept in a slanted position overnight to allow serum separation. The separated serum samples were transferred into sterile 1.8 ml cryovial tubes and stored in a deep freezer at -20°C in the Laboratory of Animal Health Research Division, Nepal Agricultural Research Council, Khumaltar, Lalitpur, until serological analysis. The serum samples were examined for the detection of specific antibodies against Mccp by using the C-ELISA test kit manufactured by IDEXX (CIRAD-Montpellier, France) containing the monoclonal anti-Mccp antibody Mab 4.52 in pre-coated plates with ready-made reagents. The assay was conducted according to the test protocol supplied by the kit.

Data management and analysis

The collected data in the field along with serological results obtained from laboratory analysis were entered in a Microsoft Office Excel spreadsheet. The entered data in Microsoft Office Excel spreadsheet were imported into SPSS V 20.0 to perform statistical analysis. The collected data were calculated by using descriptive statistics and inferential statistics. The overall seroprevalence was calculated by dividing the number of seropositive goats by the total number of goats tested. Chi-square test and fisher’s exact test were used to detect the association between seropositivity and questionnaire variables. For the analysis, a confidence level of 95% and a p-value of < 0.05 were considered to determine statistical significance.

Ethical statement

Ethical approval of the study was sought from the internship advisory committee, Veterinary Teaching Hospital, Paklihawa Campus, Institute of Agriculture and Animal Science and Animal Health Research Division, Nepal Agricultural Research Council (NARC) during the proposal seminar. Verbal consent was obtained from the respondent by explaining the objectives of the survey. Blood was collected by a technical expert with minimal pain to the animals.

Results and Discussion

Among the total 89 serum samples tested by cELISA, 3 serum samples were found seropositive for specific antibodies against Mycoplasma capricolum capripneumoniae with an overall apparent seroprevalence of 3.37% (1.15±9.45). Assuming the sensitivity and specificity of the ELISA kit to be 1 and 0.99, the true prevalence of the study was calculated via Epitools- Epidemiological calculator by Ausvet (Matope et al., 2011; Peyraud et al., 2014). The true prevalence of the study was 3.4% (0.93±9.49). Among the two rural municipalities, the seropositivity for CCPP was found slightly high in Tinau than in Rohini.

The association of factors with seropositivity of CCPP was identified by using the chi-square test and fisher’s exact test whenever necessary to find out the p-value (Table 1). The results obtained after analysis showed that host-related factors such as age and sex of the goats were not significantly associated with CCPP seroprevalence (p >0.05). However, the seropositivity was slightly higher in young (4.76 %) and male goats (4.00%) compared to adult (2.94%) and female goats (3.13%), respectively. The environment-related factors; altitude and districts, were also analysed but the seroprevalence was not significantly associated with the altitude and district. Similarly, the seroprevalence was not significantly associated with farm type, breed, and pregnancy of animals (Table 1). The seroprevalence of CCPP was significantly associated with ‘history of respiratory symptoms’ (p = 0.01).

 

Table 1: Association of seropositivity with questionnaire variables in Rupandehi and Palpa districts of Nepal.

Sample categories		Positive (%)	Negative (%)	Total	P value
Districts	Rupandehi	1 (3.33%)	29(96.67%)	30	0.71
	Palpa	2 (3.39%)	57(96.61%)	59
Husbandry practices	Intensive	3 (6.38%)	44(93.62%)	47	0.14
	Semi intensive	0 (0.00%)	42(100%)	42
Age group	Young	1 (4.76%)	20(95.24%)	21	0.55
	Adult	2 (2.94%)	66(97.06%)	68
Sex	Male	1 (4.00%)	24(96.00%)	25	0.63
	Female	2 (3.13%)	62(96.88%)	64
Breed	Khari (local breed)	2 (6.67%)	28(9.33%)	30	0.26
	Others	1 (1.69%)	58(98.31%)	59
Pregnant	Yes	1 (3.85%)	25(96.15%)	26	0.65
	No	2 (3.17%)	61(96.83%)	63
Respiratory signs	Present	3(15.00%)	17(85.00%)	20	0.01*
	Absent	0 (0.00%)	69(100%)	69

 

To the best of our knowledge, this is the first study conducted to determine the seroprevalence of CCPP and associated risk factors in Nepal. The overall apparent prevalence was found to be 3.37% which is lower in comparison to the findings of (Udit and Chand, 2008) i.e., 10.5% in Uttar Pradesh state of India. Although Nepal is considered a CCPP free country, this study portrait the high risk of CCPP infection among sheep and goats in Nepal. This study also hints the presence of CCPP among goats and sheep in other parts of Nepal, which may have gone undiagnosed due to lack of diagnostic facilities, high cost, and difficulty in isolation, culture, and identification of the organism. The seropositivity of CCPP in our study may be due to the porous nature of the international borders of the country. Hence there is a need for cross-border harmonization of disease control programs including regular investigation, vaccination, strict quarantine, and transboundary animal movement regulation (Kipronoh et al., 2016; Parajuli and Acharya, 2019).

In contrast to this study, higher seroprevalences were reported in a different state of India like 33.67 % in Nagpur (Ingle et al., 2008), 10.65 % in Jabalpur, Madhya Pradesh (Gupta et al., 2016), 9.93% in Ladakh (Parray et al., 2019), 16.05 %, and 20.24 % in sheep and goats respectively in Maharashtra (Suryawanshi et al., 2015). Very low seroprevalence in our study could be due to the fact that Nepal is CCPP free country. Also, the difference in prevalence might be due to spatial and geographical variations and differences in total population, sample size, sampling techniques, and testing procedure and criteria. Our study suggests that age and sex have no role in CCPP epidemiology. In contrast to our findings, a study in Tanzania showed the seroprevalence of CCPP to be significantly (P < 0.05) higher in adults and male goats (P<0.05) (Nyanja et al., 2013). In another study in the Afar region of Ethiopia, age and sex were significantly associated with the prevalence of CCPP (Regassa et al., 2010). A study in Egypt also found that age >4 years of a goat is an important risk factor of CCPP (Selim et al., 2021). Another study in Himalayan pashmina goats in Ladakh, India reported a significant association (P≤0.05) of seropositivity with age (Parray et al., 2019). Significant association of CCPP with adult goats may be due to fact that older animals are at risk of exposure for a long period. All seropositive goats were raised in an intensive housing system but the housing system was not significant for seropositivity.

Severe respiratory distress, sero-mucoid nasal discharge, dyspnoea, coughing, and pyrexia are characteristic symptoms of CCPP (Iqbal et al., 2019). The history of respiratory symptoms in goats was significantly associated with the seropositivity of CCPP in our study. This suggests that goats might have been infected with CCPP at some stage of their life. The government of Nepal, regional and local laboratories, animal health workers, and related stakeholders should actively participate in the establishment of diagnostic (molecular and culture) facilities throughout the country. An increase in the number of prevalence studies with a large sample size, awareness of the disease with its economic importance among goat farmers, and effective governmental strategies to identify and manage the disease are urgently needed to prevent the potential economic loss.

Conclusions and Recommendations

Although Nepal is considered a CCPP free country, this study indicated the presence of CCPP in the goats of Rupandehi and Palpa districts of Nepal. The history of respiratory symptoms among the goats was significantly associated with seropositivity of CCPP which provides an additional clue about the incidence of disease in Nepal. Our study provides preliminary data on CCPP among goats in Nepal which can be the baseline for understanding the epidemiology of CCPP in Nepal. Further epidemiological studies with appropriate sample size covering different parts of the country can provide crucial insights into the prevalence of the disease in Nepal.

Acknowledgments

Authors acknowledge Animal Health Research Division (AHRD), Nepal Agricultural Research Council (NARC), for providing necessary laboratory facility to perform ELISA test. We would like to thank all the farmer who agreed to participate in the survey and allow to collect the sample from their animals.

Novelty Statement

This is the first-ever report of serological detection of CCPP antibodies in the goat population of Nepal.

Author’s Contribution

Conceptualization: Basanta Kumar Adhikari, Chet Narayan Kharel, Krishna Kaphle and Doj Raj Khanal.

Methodology: Basanta Kumar Adhikari, Deepak Subedi and Sumit Jyoti.

Project administration: Basanta Kumar Adhikari, Deepak Subedi and Sumit Jyoti.

Writing original draft: Basanta Kumar Adhikari, Deepak Subedi and Sumit Jyoti.

Writing review editing: Deepak Subedi, Krishna Kaphle, Chet Narayan Kharel and Doj Raj Khanal.

Conflict of interest

The authors have declared no conflict of interest.

References

Ahaduzzaman, M., 2020. Contagious caprine pleuropneumonia (CCPP): A systematic review and meta-analysis of the prevalence in sheep and goats. Transbound. Emerg. Dis., pp. 13794. https://doi.org/10.1111/tbed.13794

Arif, A., Schulz, J., Thiaucourt, F., Taha, A., and Hammer, S., 2007. Contagious caprine pleuropneumonia outbreak in captive wild ungulates at Al-Wabra Wildlife Preservation, State of Qatar. J. Zoo Wildl. Med., 38(1): 93–96. https://doi.org/10.1638/05-097.1

Awan, M.A., Abbas, F., Yasinzai, M., Nicholas, R.A.J., Babar, S., Ayling, R.D., Attique, M.A., Ahmed, Z., Wadood, A., and Khan, F.A., 2010. First report on the molecular prevalence of Mycoplasma capricolum subspecies capripneumoniae (Mccp) in goats the cause of contagious caprine pleuropneumonia (CCPP) in Balochistan province of Pakistan. Mol. Biol. Rep., 37(7); 3401–3406. https://doi.org/10.1007/s11033-009-9929-0

Chu, Y., Gao, P., Zhao, P., He, Y., Liao, N., Jackman, S., Zhao, Y., Birol, I., Duan, X., and Lu, Z., 2011. Genome sequence of mycoplasma capricolum subsp. capripneumoniae strain M1601. J. Bacteriol., Am. Soc. Microbiol. J., 193(21): 6098–6099. https://doi.org/10.1128/JB.05980-11

Gupta, D.K., Shukla, P.C., Tiwari, A., Baghel, R.P.S., Sharma, V., Shivhare, J., and Gupta, N., 2016. Seroprevalence Study on Goat Contagious Caprine Pleuropneumonia in Jabalpur, Madhya Pradesh. J. Anim. Res., 6(4): 743. https://doi.org/10.5958/2277-940X.2016.00092.9

Hussain, R., Auon, M., Khan, A., Khan, M.Z., Mahmood, F., and Ur-Rehman, S., 2012. Contagious caprine pleuropneumonia in Beetal goats. Trop. Anim. Health Prod., 44(3): 477–481. https://doi.org/10.1007/s11250-011-9922-1

IDRC, 2016. Contagious caprine pleuropneumonia. Monograph 03. https://idl-bnc-idrc.dspacedirect.org/bitstream/handle/10625/58278/IDL-58278.pdf

Ingle, V.C., Sivakumar, P., Kalorey, D.R., Pote, D.E., Dhamanna-Patil, P.S., Chavhan, S.K., Nagdive, A.A., and Hatkar, D.N., 2008. Seroprevalence of contagious caprine pleuropneumonia in goats in Nagpur district of Vidarbha region. Vet. World, 1(9): 270–271.

Iqbal, Y.M., Raffiq, P.O., Tauseef, B.S., Muheet, Ahmed, B.R., Gopalakrishnan, A., Karthik, K., Dhama, K., and Vir Singh, S., 2019. Contagious caprine pleuropneumonia a comprehensive review. Taylor and Francis Ltd. Vet. Quart., 39(1): 1–25. https://doi.org/10.1080/01652176.2019.1580826

Jain, U., Verma, A.K., and B.C.P., 2015. Occurrence of mycoplasma infection in barbari goats of Uttar Pradesh, India. Haryana Vet., 54(1): 53–55.

Jores, J., Baldwin, C., Blanchard, A., Browning, G.F., Colston, A., Gerdts, V., Goovaerts, D., Heller, M., Juleff, N., Labroussaa, F., Liljander, A., Muuka, G., Nene, V., Nir-Paz, R., Sacchini, F., Summerfield, A., Thiaucourt, F., Unger, H., Vashee, S., and Salt, J., 2020. Contagious bovine and caprine pleuropneumonia: A research community’s recommendations for the development of better vaccines. NPJ Vaccines Nat. Res., 5(1): 1–9. https://doi.org/10.1038/s41541-020-00214-2

Kipronoh, K.A., Ombui, J.N., Binepal, Y.S., Wesonga, H.O., Gitonga, E.K., Thuranira, E., and Kiara, H.K., 2016. Risk factors associated with contagious caprine pleuro-pneumonia in goats in pastoral areas in the Rift Valley region of Kenya. Preventive Vet. Med., 132: 107–112. https://doi.org/10.1016/j.prevetmed.2016.08.011

Matope, G., Muma, J.B., Toft, N., Gori, E., Lund, A., Nielsen, K., and Skjerve, E., 2011. Evaluation of sensitivity and specificity of RBT, c-ELISA and fluorescence polarisation assay for diagnosis of brucellosis in cattle using latent class analysis. Vet. Immunol. Immunopathol., 141(1–2): 58–63. https://doi.org/10.1016/j.vetimm.2011.02.005

MOLD, 2017. Livestock Statistics of Nepal (Issue July). Ministry of Livestock Development, Government of Nepal. www.mold.gov.np

Nyanja, P.M., Kusiluka, L.J.M., and Mellau, S.B., 2013. Prevalence of contagious caprine pleuropneumonia in goats in Musoma District of Mara Region, Tanzania. Tanzania Vet. J., 28(1). https://www.ajol.info/index.php/tvj/article/view/98551

OIE, 2009. Contagious caprine pleuropneumonia. https://www.oie.int/app/uploads/2021/03/contagious-caprine-pleuro.pdf

Parajuli, S., and Acharya, K.P., 2019. Detection of antibodies against bluetongue virus among goats of different eco-climatic zones of Nepal. Comp. Clin. Pathol., 28(4): 1177–1180. https://doi.org/10.1007/s00580-019-02992-1

Parray, O.R., Yatoo, M.I., Muheet, Ahmed, B.R., Ullah, M.H., Bashir, S.T., and Nabi, M.S., 2019. Seroepidemiology and risk factor analysis of contagious caprine pleuropneumonia in Himalayan Pashmina Goats. Small Rumin. Res., 171: 23–36. https://doi.org/10.1016/j.smallrumres.2018.12.004

Peyraud, A., Poumarat, F., Tardy, F., Manso-Silván, L., Hamroev, K., Tilloev, T., Amirbekov, M., Tounkara, K., Bodjo, C., Wesonga, H., Nkando, I. G., Jenberie, S., Yami, M., Cardinale, E., Meenowa, D., Jaumally, M. R., Yaqub, T., Shabbir, M.Z., Mukhtar, N., and Thiaucourt, F., 2014. An international collaborative study to determine the prevalence of contagious caprine pleuropneumonia by monoclonal antibody-based cELISA. BMC Vet. Res., pp. 10. https://doi.org/10.1186/1746-6148-10-48

Regassa, F., Netsere, M., and Tsertse, T., 2010. Sero-prevalence of contagious caprine pleuropneu-monia in goat at selected woredas of Afar Region. Ethiopian Vet. J., 14(1).

Selim, A., Megahed, A., Kandeel, S., Alanazi, A.D., and Almohammed, H.I., 2021. Determination of seroprevalence of contagious caprine pleuropneumonia and associated risk factors in goats and sheep using classification and regression tree. Animals, 11(4): 1165. https://doi.org/10.3390/ani11041165

Sergeant, E., 2018. Epitools epidemiological calculators. Ausvet. Ausvet. http://epitools.ausvet.com.au.

Suryawanshi, S.N., Tembhurne, P.A., Gohain, S., Kesharkar, J.A., Tumlam, U.M., and Ingle, V.C., 2015. Seroprevalence of contagious caprine pleuropneumonia in small ruminants in Maharashtra. Indian J. Vet. Sci. Biotechnol., 10(4): 73–74.

Thiaucourt, F., Bölske, G., Leneguersh, B., Smith, D., and Wesonga, H., 1996. Diagnosis and control of contagious caprine pleuropneumonia. OIE Revue Sci. Tech., 15(4): 1415–1429. https://doi.org/10.20506/rst.15.4.989

Udit, J., and Chand, B.P., 2008. Epidemiology of Mycoplasma pneumonia in goats of India. International Organization for Mycoplasmology. 17th International Congress. July 6-11, 2008, Tianjin Medical University, China, pp. 67

Wazir, I., Hussain, I., Khan, M.A., Ali, M.I., Rahman, H.U., Ashraf, F., Khan, S., Khan, B., Ullah, S., Ullah, Q., and Khan, A., 2016. Seroepidemiological analysis of contagious caprine pleuropneumonia through cELISA in selected districts of Khyber Pakhtunkhwa, Pakistan. Am. Sci. Res. J. Eng. Technol. Sci., 26(3): 274–281. http://asrjetsjournal.org/

Yu, Z., Wang, T., Sun, H., Xia, Z., Zhang, K., Chu, D., Xu, Y., Xin, Y., Xu, W., Cheng, K., Zheng, X., Huang, G., Zhao, Y., Yang, S., Gao, Y., and Xia, X., 2013. Contagious caprine pleuropneumonia in endangered Tibetan Antelope, China, 2012. Emerg. Infect. Dis. CentersDis. Contr. Prevent., 19(12): 2052–2053. https://doi.org/10.3201/eid1912.130067