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Rickettsial and Haemoparasitic Infections in Dogs of Malwa Region, Madhya Pradesh - An Etiopathological Study.

Introduction

Canines are known to be infected by different haemoparasites viz. Babesia spp., Trypanosoma spp., Leishmania spp., Hepatozoon spp., Ehrlichia spp. and Mycoplasma spp. (Haemobartonella) transmitted through different arthropod vectors like ticks, lice, triatomines, mosquitoes, tabanids and phlebotomine sand flies to produce illness collectively termed as Canine Vector Borne Diseases (CVBD) in tropical and sub-tropical countries including India (Bhattacharjee and Sarmah, 2013) are minimal. In addition, deforestation has changed the natural habitation of vectors and introduced newer vectors from wildlife into rural and urban areas. Clinical findings in vector borne diseases range from incidental hematological changes to severe life threatening illness due to synergistic pathological effects between etiological agents. This complicates diagnosis, treatment and can adversely influences prognosis if the practitioner fails to suspect, document and treat each concomitant infection (Sarma, 2015).

Material and Methods

For epidemiological study, a total of 150 dogs of either sex and various breeds covering hot and humid weathers with clinical signs of constant and relapsing fever and lymph adenopathy were examined over a period of twelve months from i.e. March' 2016 to February' 2017. 3 ml of blood was collected in a vial containing EDTA (anticoagulant) @ 2 mg/ml of blood from cephalic or saphenous vein. Blood smears were prepared on clean grease free glass slides and stained with Wright-Giemsa Stain (modified wright's stain) (Benjamin, 2007) and Knott's method for demonstration of microfilariae (Sloss et al., 1994). The parasites were identified on the basis of characteristic morphology. The data obtained were analysed using Chi-square test.

Results and Discussion

Out of 150 adult dogs of both sex and mixed breeds, 47 (31.33%) dogs were found positive for rickettsial, haemoparasitic and mixed infections. The incidence of rickettsial infection was found to be highest in Malwa region viz.17.33 percent with maximum number of Ehrlichia spp. positive cases, followed by haemoparasitic (9.33%) and mixed infections (4.66%) (Table 1).

This incidence is in agreement with findings of Dhankar et al. (2011) who mentioned in their study about more infection of Ehrlichiosis than other parasitic infections and are in contrast, with the findings of Tsegay et al. (2016) who noted more number of other parasitic infections compared to Ehrlichiosis. According to Ybanez et al. (2016) pathogens transmitted by same tick vector, included Anaplasma platys and Babesia gibsoni. Ticks have known capability of hosting multiple pathogenic organisms, which can result in co-infected dogs.

Concommitant infection of E canis with H. canis and also E. canis with Babesia spp. were earlier documented by Sharma et al. (2015).

The maximum incidence of rickettsial and haemoparasitic infections was noticed in German Shepherd breed i.e. 4.6 percent and 4.6 percent respectively (Table 2).

These findings match with Shrivastava and Shukla (2013) who reported higher incidence of rickettsial and haemoparasitic infections in hairy breed dogs, which may be due to more tick infestation and difficulties during tick control because of their long hair coat. According to Tsegay et al. (2016), German shepherd dog is more prone to develop more severe clinical signs to haemoparasites due to the fact that cell-mediated immunity was found reduced in such infections and is in contrast with the finding of Kottadamane et al. (2017) who reported highest prevalence in Labrador retriever. The male dogs revealed higher risk of infection as compared to female dogs (Table 3).

These findings were similar to the observations of Sahu et al. (2014). The higher incidence can be attributed to hormonal factors or frequent roaming behaviour of males in search of mates and territory (Tsegay et al., 2016). The season wise incidence of rickettsial and haemoparasites are presented in Table 4. The higher incidence of rickettsial and haemoparasites were recorded during June end to October 8.6 percent rickettsial, 5.3 percent haemoparasitic and 2.6 percent mixed infection. Followed by the period between November-February and least incidence was record during March-mid June Infections at the end of summer, rainy and initial months of winter was possibly due to higher density of vector population in these months.

These findings were in accordance with Nalubamba et al. (2011). Most animals suffered during monsoon months, might be due to more ticks in monsoon that had developed during summer months. This is in accordance with Kottadamane et al. (2017) who observed more prevalence in rainy and summer followed by autumn and least in spring season. The probable reason behind the trend may be correlated to seasonal activity of brown dog tick, R. sanguineus which was more abundant in hot and humid period of year and are dissimilar to the results recorded by Sahu et al. (2014) who reported that disease is spread by ticks prevalence of which is also high during summer.

In this study, seven species of rickettsia and haemoparasites viz. Ehrlichia canis 16 (10.66%) (Fig.1), Ehrlichiae ewingii 4 (2.6%) (Fig. 2), Ehrlichia platys 10 (6.6%) (Fig. 3), Babesia gibsoni 8 (5.3%) (Fig. 4), Babesia canis 5 (3.33%) (Fig. 5), Hepatozoon canis 2 (1.33%) (Fig. 6) and Trypanosoma evansi 2 (1.33%) (Fig. 7) either as single or mixed infection was recorded. Dirofilaria immitis (microfilaria) and Leishmaniosis was not detected during the study period (Table 5).

In present study, Ehrlichia was observed as intracytoplasmic bodies of varying size and shapes in monocytes, lymphocytes, neutrophils and platelets. Granulocytic ehrlichia were detected as morulae or inclusion bodies in granulocytes mainly in neutrophils. In haemoparasitic infections, Babesia spp. was visible within the red blood cells and these were of lightly basophilic pyriform shape with indistinct internal structures. Blood smears revealed ellipsoid, elongated, pale staining cytoplasmic bodies with one centrally located blue nucleus. These inclusion bodies were identified as gamatocytes based on their morphological characteristics as in Hepatozoon canis and blood smears revealed numerous free extra cytoplasmic protozoa. Two cases were found positive for Trypanosoma evansi. Trypanosoma evansi is a monomorphic trypanosome but polymorphism may also be seen. Results were comparable with the findings of Dhankar et al. (2011) and Ybanez et al. (2016).

Conclusion

Incidence of varying degrees of rickettsial and haemoparasitic infections were 31.33% in blood samples of dogs. Cases of rickettsia (17.33%), haemoparasitic (9.33%) and mixed infections (4.66%) were determined and differentiated. Cases of rickettsial infection were found to be highest. Highest incidence was recorded in rainy season. The most susceptible breed was German Shepherd. Male dogs of all breeds were affected more than female dogs.

Acknowledgements

Authors are thankful to the Dean, Veterinary College Mhow, Head and Staff of Department of Pathology for providing necessary facilities to conduct the research work.

References

Benjamin, M.M. (2007). Outline of Veterinary Clinical Pathology, 3rd Edn., Kalyani Publishers, New Delhi, p. 30.

Bhattacharjee, K. and Sarmah, P.C. (2013). Prevalence of haemoparasites in pet, working and stray dogs of Assam and North-East India - A Hospital based study. Vet. World 6: 874-78.

Dhankar, S., Sharma, R.D. and Jindal, N. (2011). Some epidemiological observation on canine ehrlichiosis in Haryana and Delhi state. Haryana Vet. 50: 9-14.

Kottadamane, M.R., Dhaliwal, P.S. and Singla, L.D. (2017). Diagnosis and treatment of canine monocytic ehrlichiosis in a boxer breed of dog - A Case Report. Internat'l J. Sci. Enviro.Technol. 5: 3099-3105.

Nalubamba, K.S., Hankanga, C, Mudenda, N.B. and Masuku, M. (2011).The epidemiology of canine Babesia infections in Zambia. Prev. Vet. Med. 99: 240-44.

Sahu, A., Mohanty, B., Panda, M.R. and Sardar, K.K. (2014). Incidence of haemoprotozoan parasites in dogs in and around Bhubaneswar, Odisha. Indian Vet. J. 91: 93-95.

Sarma, K., Mondal, D.B., Saravanam, M. and Mahendra, K. (2015). Evaluation of haemato-biochemical and oxidative indices in naturally infected concomitant tick borne intracellular diseases in dogs. Asian Pacific J. Tropical Disease 5: 60-66.

Sharma, D.K., Gupta, V.K., Bansal, S., Joshi, V., Mandal, R.S.K., Singh, M. and Bhanuprakash, A.G. (2015). Therapeutic efficacy of Doxycycline with whole blood transfusion in management of thrombocytopenic ehrlichiosis in canines. Internat'l J. Adv. Res. 3: 353-57.

Shrivastava, S. and Shukla, P.C. (2013). Prevalence of canine babesiosis in dogs at and around Jabalpur (M.P.). Bioinfolet 10: 905-06.

Sloss, M.W., Kemp. R.L. and Zajac, A.M. (1994). Veterinary Clinical Parasitology, 6th Edn., International Book Distributing Co., Lucknow, p. 107-08.

Tsegay, A.K. Abebe, B. Amano, F. and Gemeda, A. (2016). Study on Prevalence of major tick and tick borne hemoparasites of dogs visiting Jimma University Veterinary Open Air Clinic. Middle-East J. Scientific Res. 24: 2342-51.

Ybanez, A.P., Ybanez, R.H.D., Villavelez, R.R., Malingin, H.P.F., Barrameda, D.N.M., Naquila, S.V. and Olimpos, S.M.B. (2016). Retrospective analyses of dogs found serologically positive for Ehrlichia canis in Cebu, Philippines from 2003 to 2014. Vet. World 9: 43-47.

Pramila Kirade, S. Shukla (1), N. Shrivastava, S.D. Audarya and G.P. Jatav

Department of Veterinary Pathology

College of Veterinary Science and Animal Husbandry

Nanaji Deshmukh Veterinary Science University (NDVSU)

Mhow - 453446 (Madhya Pradesh)

(1.) Professor and Corresponding author.

E-mail: drsupriyav@gmail.com
Table 1: Incidence of rickettsial and haemoparasitic infections in dogs

                 Number of   Incidence
Infections       positive    (%)
                 cases

Rickettsial      26          17.33
Haemoparasitic   14           9.33
Mixed infection  07           4.66
Total            47          31.32

Table 2: Breed-wise incidence of rickettsial and haemoparasitic
infections (n=47)

Breed            Rickettsial  Haemo-      Mixed
                              parasities  infection

Rottweiler        1.33% (2)   -           -
Great Dane        1.33%(2)    -           -
Labrador          4% (6)      4% (6)      0.66% (1)
German Shepherd   4.6% (7)    4.6% (7)    -
New Foundland     0.66%(1)    -           -
Shih-Tzu          0.66% (1)   -           -
Pomeranian        1.33% (2)   -           -
Desi              3.3% (5)    0.66% (1)   4% (6)
Total            17.33% (26)  9.33% (14)  4.66% (7)

[X.sup.2] = 24.54 (*); ((*) P<0.05) Significant at 5% level of
significance, n= number in parenthesis

Table 3: Gender-wise incidence of rickettsial and haemoparasitic
infections

Gender           Male        Female

Rickettsial      12.6% (19)  4.6% (7)
Haemoparasitic    6.6% (10)  2.66% (4)
Mixed infection   2.6% (4)   2% (3)

[X.sup.2] = [1.062.sup.NS]; NS=Non significant, Chi square is found to
be non significant at 5% level of significance, n= number in
parenthesis

Table 4: Season-wise incidence of rickettsial and haemoparasitic
infections

Group            March-    June-      November-
                 mid       October    February
                 June

Rickettsial      2% (3)    8.6% (13)  6.6% (10)
Haemoparasitic   0.6% (1)  5.3% (8)   3.3% (5)
Mixed infection  -         2.6% (4)   2% (3)

[X.sup.2] = [2.590.sup.NS]; NS= Non significant, Chi square is found to
be non significant at 5% level of significance, n= number in parenthesis

Table 5: Species-wise incidence of rickettsial and haemoparasitic
infections

Parasite species      Single         Mixed          Total
                      infection (%)  infection (%)  %

Ehrlichia canis       9.3 (14)       1.33 (2)       10.66 (16)
Ehrlichiae ewingii    2.6 (4)        -               2.6 (4)
Ehrlichia platys      5.5 (8)        1.33 (2)        6.6 (10)
Babesia gibsoni       4.6 (7)        0.66 (1)        5.5 (8)
Babesia canis         3.33 (5)       -               3.33 (5)
Trypanosoma evansi    0.66 (1)       0.66 (1)        1.33 (2)
Hepatozoon canis      0.66 (1)       0.66 (1)        1.33 (2)
Dirofilaria immitis   Nil            Nil            Nil
(microfilaria)
Leishmaniosis         Nil            Nil            Nil
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Article Details
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Title Annotation:Research Article
Author:Kirade, Pramila; Shukla, S.; Shrivastava, N.; Audarya, S.D.; Jatav, G.P.
Publication:Intas Polivet
Article Type:Report
Geographic Code:9INDI
Date:Jan 1, 2018
Words:1852
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