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SERO-EPIDEMIOLOGICAL ANALYSIS OF CONTAGIOUS CAPRINE PLEUROPNEUMONIA IN GOATS.

Byline: M. A. Chandio, D. H. Kalhoro, S. H. Abro, M. S. Kalhoro, A. Kaka, G. M. Lochi, A. A. Soomro, M. A. Nizamani and A. A. Chandio

Keywords: Contagious caprine pleuropneumonia, Enzyme Linked Immuno-Sorbent Assay, Capri-Latex Agglutination Test, Goat, Sindh, Pakistan.

INTRODUCTION

The small ruminant faces serious constraints in form of inadequate nutrition, low genetic potential, intense harsh climatic conditions, poor management and various diseases. Contagious Caprine Pleuropneumonia (CCPP) is a devastating threat to the small ruminants with major economic importance (Awan et al., 2009). CCPP is a lethal respiratory disease having serious impact in reduction of small ruminants' production, as the fastidious, intracellular bacteria infect domestic as well as wild breeds of small ruminants with high morbidity (100%) and mortality rates may reach up to (60-70%) (Ostrowski et al., 2011). All age groups and both sexes were susceptible but high mortality was recorded in young kids as compared to adults (Arif et al., 2007). The CCPP is caused by Mycoplasma capricolum sub-specie capripneumoniae (Mccp) (OIE 2012). It belongs to the genus Mycoplasma and has a very short incubation period of 3-5 days (Thiaucourt and Bolske 1996).

The typical clinical signs of CCPP are hyperpyrexia (41-43 AdegC), after an onset of high fever, respiratory symptoms become more pronounced with increased nasal discharge and lacrimation (OIE, 2009). Other clinical signs such as lameness, diarrhea, inability to move, unappropriated gait with stiff neck and prostration on ground with lateral recumbency can be observed in advance stages of the disease (OIE, 2008). Analyzing the seroprevalence provides a scientific knowledge about the occurrence of disease in a particular area, to better understand about its risk factors such as age, sex, breed, specie and its seasonally outbreaks, as it also provides a baseline data for the progressive development of future strategies for diagnosis and control of certain highly contagious diseases. Highlighting the studies on seroprevalence of CCPP in different areas with variation in results such as, overall seroprevalence of 18.61% was observed in southern Ethiopia (Mekuria and Asmare 2009).

Higher seroprevalance of 32.5% was recorded in Beetal goat breed in Pakistan (Hussain et al., 2012) and 37.5% was documented in East Turkey (Cetinkaya et al., 2009). On the contrary, Mekuria et al., (2008) reported a lower seroprevalence of 15.5% in a participatory investigation of CCPP in the Benna-Tsemay and Hammer districts of southern Ethiopia.

Gross pathological lesions of CCPP are limited exclusively to lung and pleura. Affected lungs shows necrotic multi focal and sometime diffused lesions having a port wine color (Thiaucourt and Bolske 1996). Swelling of Bronchial and mediastinal lymph nodes with plural adhesions on the wall of chest cavity are common features of the disease. Hemorrhagic lesions and hypertrophy can also be visualized on liver and kidneys (Gelagay et al., 2007; Mondal et al., 2004). Hemorrhages can be easily seen in tracheal; with sloughing of the epithelial layers of the lung tissues may show acute serofibrinous condition to the chronic fibrino-necrotic pleuropneumonia (Hernandez et al., 2006).

There are different techniques used for diagnostic purpose during CCPP outbreaks. The ELISA (Enzyme Linked Immunosorbent Assay) and the latex agglutination test (LAT) play an important role in identifying animals suffering from contagious caprine pleuropneumonia (OIE, 2009). Properties like low cost, easy to use and sensitivity increases its worth in detection of disease in outbreaks. The competitive ELISA and LAT are more sensitive than biochemical tests (Thiaucourt et al.,1994).

In spite of aforementioned prevailing situation with an existence of the numerous problems due to the CCPP, there is lack of well documented scientific information on the occurrence of CCPP among goats in Sindh, especially in district Hyderabad and Tando Allahyar. Therefore, this study was designed with the aim to determine the seroprevalence of CCPP in Hyderabad and Tando Allahyar districts of Sindh province, and to assess the potential risk factors for the occurrence of disease.

MATERIALS AND METHODS

Study Area: For the present study, a project was designed to record Sero-epidemiological status and pathogenesis of contagious caprine pleuropneumonia in District Hyderabad and Tando Allahyar. The study area was divided into Rural and Urban sectors of Hyderabad and Tando Allahyar, involving randomly selection of Government and private farms. To address the stated objectives, semi-structured questionnaires was managed to select individual farms in study areas to gather information of risk factors such as age (1, 2, 3 and 4 years), sex and Tapri, Kamori, Tedi and Pateri breeds for CCPP occurrence in the area.

Collection of Samples: For the determination of prevalence rate, a total of 200 blood samples were collected during October 2016-March 2017 from goats showing clinical signs of pneumonia, with the suspected CCPP infection.

Collection of blood Samples: About 3 ml of blood was aseptically collected from goats in a clot activator vacutainer. Samples were transported in a cold chain to the Central Veterinary Diagnostic Laboratory Tandojam for the further investigation. Once the serum was separated by centrifugation, it was either immediately used or stored in deep freezer at -20AdegC till screened.

Processing of serum samples: Serum samples were thawed at the room temperature before analyzed by Latex Agglutination Test kit (Capri-LAT) and C-ELISA kit (Competitive Enzyme Linked Immunosorbent Assay) using three replicates for confirmation of disease as prescribed by (OIE, 2009).

Latex Agglutination Test: Capri-LAT prepared reagent was brought from the Animal and Plant Health Agency-Weybridge, United Kingdom, donated by Central Veterinary Diagnostic Laboratory Tandojam under the project of 'Strengthening of Laboratories in identification and eradication of CCPP from Sindh". The test procedure was followed as prescribed by manufacturer and adopted by (Shahzad et al., 2012).

Competitive Enzyme Linked Immunosorbent Assay: The samples were also analysed for the detection of Mycoplasma capricolum subsp. capripneumoniae (Mccp) using monoclonal antibody based competitive C-ELISA Test Kit (IDEXX CCPP, 0656231-01). The 1st batch of C-ELISA Kit was developed at CCPP reference laboratory CIRAD-Montpellier France, purchased under the research project "'Strengthening of Laboratories in identification and eradication of CCPP from Sindh, at Central Veterinary Diagnostic Laboratory Tandojam. The serum samples were processed according to CCPP C-ELISA kit.

Sampling for isolation: For the isolation of Mycoplasma capricolum sub-specie capripneumonie, sampling from pleural fluids and lung tissues are required as prescribed by OIE (2008), for this purpose postmortem was performed on dead animals during disease outbreaks. Tissue samples were taken in sterile container containing 10% formalin with proper labelling. Pleural swabs were aseptically taken and inserted in test tube containing modified hay flicked medium, after proper sampling each tube was labelled and transported under proper refrigeration (4AdegC) to CVDL Tandojam.

Growth Medium for Mycoplasma: Modified hay flick culture medium is well known to be used for cultivation of Mycolplasma. The standards were followed as mentioned by (OIE, 2008). Both agar and broth mediums were prepared as suggested by manufacturers. Samples were inoculated and streaked according to standard operating procedure (OIE, 2009).

Tissue Sampling: Tissue samples collected from the lungs and trachea were fixed in 10% formalin. The standard protocol of HandE technique was followed by tissue sectioning, and staining for detailed histopathological examination as suggested by Wesonga et al., (2004).

Table 1. The overall seroprevalence of CCPP among goats in districts Hyderabad and Tando Allahyar analysed by different techniques.

###Districts

Tests###Hyderabad###Tando Allahyar###Overall prevalence###P-Value

###Prevalence###Prevalence###No.(%)

###No.(%)###No.(%)

Capri-LAT###24(24.00)###20(20.00)###44(22.00)###0.495

C-ELISA###20(20.00)###17(17.00)###37(18.50)###0.585

P-Value###0.495###0.585###0.313###-

Table 2. Prevalence of CCPP among goats in Districts Hyderabad and Tando Allahyar in relation to the various risk factors analyzed by Capri-LAT.

###Districts

Risk Factors###Hyderabad###Tando Allahyar

###Total###Prevalence###P-Value###Total###Prevalence###P-Value

###Samples###No.(%)###Samples###No.(%)

Sex###Male###11###02(18.18)###0.632###09###01(11.11)###0.485

###Female###89###22(24.71)###91###19(20.87)

### 3 Yrs###36###15(41.66)###29###12(41.37)

###Tapri###17###08(47.05)###13###02(15.38)

Breed###Kamori###24###02(08.33)###0.030###25###03(12.00)###0.041

###Tedi###32###09(28.12)###33###04(12.12)

###Pateri###27###05(18.51)###29###11(37.93)

Table 3. Prevalence of CCPP among goats in Districts Hyderabad and Tando Allahyar in relation to the various risk factors analyzed by C-ELISA:

###DISTRICTS

RISK###Hyderabad###Tando Allahyar

FACTORS###Total###Prevalence###P-Value###Total###Prevalence###P-Value

###Samples###No.(%)###Samples###No.(%)

Sex###Male###11###01(09.09)###0.338###09###01(11.11)###0.622

###Female###89###19(21.34)###91###16(17.58)

Age### 3 Yrs###36###13(36.11)###31###10(32.25)

###Tapri###17###08(47.05)###13###01(07.69)

Breed###Kamori###24###01(04.16)###0.004###25###03(12.00)###0.029

###Tedi###32###08(25.00)###33###03(09.09)

###Pateri###27###03(11.11)###29###10(34.48)

RESULTS

During current sero-epidemiological study 200 serum samples were collected from different areas of both districts and examined by C-ELISA and Capri-LAT tests. To accomplish parameters of risk factor analysis, data regarding age, sex and breed were also collected. On postmortem examination of field isolated dead animals, gross lesions were recorded from lungs and trachea. Samples were also processed for bacterial isolation and histopathological examination.

Prevalence of CCPPamong goats in districts Hyderabad and Tando Allahyar using Capri-LAT and C-ELISA: Table-1 shows the overall comparative results between two tests used in the current study. The data revealed that 44 samples (22%) were found positive by Capri-LAT test and 37 (18.50%) samples were found positive on C-ELISA. However, individually 24 (24%) samples were detected positive on Capri-LAT, and 20 (20%) through C-ELISA in Hyderabad district. Whereas, 20 (20%) on Capri-LAT, and 17 (17%) through C-ELISA was recorded in Tando Allahyar district. Results of both techniques were non-significant (P>0.05). However, the differences between the positive percentages of both districts on both techniques were also found non-significant (P>0.05).

Prevalence of CCPP in relation to Sex by Capri-LAT: Among 100 samples from each district, 11 and 89 samples were collected from male and female at district Hyderabad, while for district Tando Allahyar, 09 and 91 samples were collected from male and females respectively. The results are summarized in Table-2. A total of 2 (18.18%) male and 22 (24.71%) females in Hyderabad district and 01 (11.11%) male and 19 (20.87%) females were found positive on Capri-LAT. However, statistically there was no gender wise difference (P > 0.05) at both districts.

Prevalence of CCPP in relation to Age by Capri-LAT: During present study, Age was considered as an important risk factor that may have association with the prevalence of disease among goats in districts Hyderabad and Tando Allahyar. To achieve that goal, a total of 100 random samples for both districts were divided among 4 different age groups i.e., 3 years. Samples were analyzed by Capri-LAT and are summarized in Table-2. The result shows that the prevalence in the age group of > 3 years was found higher 16 (44.44%) and 12 (41.37%) in districts Hyderabad and Tando Allahyar. Whereas, the lowest positive samples 03 (13.04%) were recorded in age group of 1-2 years at district Hyderabad, and none of animal found positive in <1-year age group in district Tando Allahyar respectively. Statistical analysis of different age groups shows significant difference (P <0.05) for both districts.

Prevalence of CCPP in relation to breed by Capri-LAT: During present study breed is another risk factor that was considered and challenged by Capri-LAT. Results are summarized in Table 2. In Hyderabad district 08 (47.05%) Tapri, 09 (28.12%) Tedi, 05 (18.51) Pateri, and 02 (08.33%) Kamori breeds were recorded positive through Capri-LAT. While, in Tando Allahyar the highest 11 (37.93%) samples were recorded for Pateri breed with lowest of 03 (12.00%) for Kamori breed respectively. The data shows statistically significant difference (P0.05) between male and females of both districts.

Prevalence of CCPP in relation to Age by C-ELISA: 100 samples from both districts were divided among 4 groups of age i.e., 3 years and were challenged by C-ELISA (Table-3). The data revealed that among all groups the highest prevalence of 13 (36.11%) and 10 (32.25%) was recorded in age group of >3 years at district Hyderabad and Tando Allahyar. While, the lowest 01 (09.09%) and 00 (00%) prevalence was detected in young animals of age group 3 years group in goats, while lower in 3 years have shown higher rate of CCPP infection as compared to young and adult animals. The prominent pathological changes (emphysema and sloughing) were observed in trachea and lungs of affected animals.

Competing interests: The authors declare that they have no competing interests.

Acknowledgements: The authors are highly thankful to Dr Parkash Dewani (Ex-Director) Central Veterinary Diagnostic Laboratory, TandoJam, Directorate of Veterinary Research and Diagnosis, Government of Sindh, Pakistan for his kind co-operation and help in collection of data and biological material.

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Author:M. A. Chandio, D. H. Kalhoro, S. H. Abro, M. S. Kalhoro, A. Kaka, G. M. Lochi, A. A. Soomro, M. A. N
Publication:Journal of Animal and Plant Sciences
Geographic Code:6ETHI
Date:Oct 31, 2019
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