prevalence of bacterial and fungal diseases in

Veterinary Practitioner Vol. 16 No. 1

June 2015

PREVALENCE OF BACTERIAL AND FUNGAL DISEASES IN DROMEDARY CAMELS IN THE RAJASTHAN STATE OF INDIA F.C. Tuteja, S.S. Dahiya and S.D. Narnaware National Research Centre on Camel, Jorbeer, Shivbari, P.B.-07, Bikaner-334 001, Rajasthan, India ABSTRACT Survey was carried out to investigate the occurrence of bacterial and fungal diseases in camels in thickly camel populated areas in semi-arid and arid climatic conditions of Rajasthan state. Amongst skin infections, fungal infections were more prevalent than the bacterial infections. Skin infections were observed more in the winter season preceding rainy season and were more prevalent in young calves of less than one year of age followed by aged (>15 yrs), active grower (1-5 yrs) and adults (5-15 yrs). A total of 32 species of various fungi were isolated and identified from camel skin infections. Mastitis was mainly caused by Staphylococc us aureus and was more prevalent in peak yielders between 6-12 year s of age. Incidence of abortion was found to increase with the advancement in the age of the animal. One serum sample from aborted field cases was positive for antibrucella antibodies, whereas all samples from pregnant animals of organised herd were negative for antibrucella antibodies. Only sporadic cases of actinobacillosis were observed and recorded. Actinobac illos is was more prevalent in summer followed by rainy and winter seasons. Actinobacillosis les ions were more prevalent in young calves and the incidence decreased with the advancement of the age of the animal. Diarrhoea was commonly observed in the rainy s eason followed by winter seas on. Fifty per cent of the diarrhoeic f aecal samples examined were positive for Gram positive Corynebacterium spp. Pasturellosis was recorded as a herd out break after heavy rain fall in the month of November. Various bacterial isolates from different infections inclu ded S. aureus, S. epidermidis, Corynebacterium spp., Pseudomonas spp., Streptococcus spp., Bacillus spp., Escherichia coli and Pasturella spp. In considering antibacterial sensitivity, tetracycline is still found to be a very effective drug for treatment of various bacterial infections in camels. Key words: Prevalence, bacterial, fungal, camel, disease

Introduction Dromedary camel is well adapted than any other domestic animal to the arid and semi-arid regions of the Rajasthan state of the India. In such regions camel not only survived during droughts, but continued producing and reproducing (Schwartz, 1992). The physiological attributes of the camel makes this animal most suitable for uses in climatic extremes (Yagil, 1985 and Wernery, 1992). In their natural habitat camels are exposed to severe stress conditions which render them susceptible to many diseases (Abbas et al., 1993 and Agab, 1993). Although camels were considered in the past, as resistant to many disease causing agents (Zaki, 1948 and Dalling et al., 1988). Now it has been realized that they are susceptible like other livestock or even more, to the common disease causing pathogens (Wilson, 1984; Abbas and Tilley, 1990 and Abbas and Agab, 2002). Among pathogenic agents bacterial infections play a major role (El-Gayown, 1986 and Margan, 1987). Only scanty information is available on skin lesions (Qureshi et al., 2002), mastitis (Tuteja et al., 2003 and Bhatt et al., 2004), actinobacillosis (Purohit et al., 1988), brucellosis (Shome et al., 2013), pasturellosis (Kachhawaha et al., 2012) in camels from India. There are reports of occurrence of various fungal skin infections in camels (Wernery and Kaaden, 2002). Therefore, based primarily on appearance of clinical signs and symptoms the present study was carried out to know the prevalence of bacterial and fungal diseases in camels in the Rajasthan state of India. Materials and Methods Survey areas: Survey was carried out to investigate the occurrence of bacterial and fungal infections in camels dwelling

in the thickly camel populated areas of semi-arid and arid climatic conditions covering districts Bikaner, Nagaur, Hanumangarh, Barmer, Jodhpur, Sirohi, Pali, Jaisalmer, Jhunjhnu, Churu and Udaipur of Rajasthan state. From July, 2007 to April, 2013, regular visits in camel rearing villages, on farm visits (also includes desert rangelands) and organized free treatment camps with the help of local veterinary officers and heads of the camel rearing Raika community were made. A total of 2248 cases were attended and information concerning the occurrence of disease along with age, sex, breed and managemental practices adopted by the farmers were recorded. Collection and processing of samples: In the affected camels with clear skin lesions (Fig. 3), ointments or other local applications present were first removed with an alcohol wipe. Then using a blunt scalpel lesions were firmly scrapped, particularly at the advancing border. If multiple lesions were present then the appropriate lesions were selected depending upon the history and clinical manifestation of the infection. From lesions with abscess formation, swab samples were collected aseptically. Similarly, from cases of diarrhoea/enteritis direct rectal swabs were collected. These samples were collected in the sterile vials and transferred to the laboratory under cold conditions. Samples were subjected to mycological and bacteriological examination by using Sabourauds dextrose chloramphenicol agar, five per cent sheep blood agar and MacConkey’s lactose agar following standard procedures. From cases of pasturellosis, blood and/or odematous fluid were collected with the help of sterile vacutainer and disposable syringe. From cases of clinical mastitis (Fig. 2),

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aseptically collected quarter milk samples were examined bacteriologically by using five per cent sheep blood agar and MacConkey’s lactose agar following standard procedures (Brown et al., 1981). From cases with history of abortion, blood samples were collected with the help of sterile vacutainer, serum was harvested and representative samples were sent for examination of antibrucella antibody titres at Project Directorate on Animal Disease Monitoring and Surveillance (PDADMAS), Bangalore. From cases showing lesions typical of actinobacillosis (Fig. 1), pus swabs collected from mature open wounds were subjected to direct smear examination. In vitro antibiotic sensitivity testing of the bacterial isolates: A total of 78 bacterial isolates comprising S. aureus (20), S. epidermidis (28), Corynebacterium spp. (18), Bacillus spp. (10), Pseudomonas spp. (2) were subjected to antibiotic sensitivity to six antimicrobials by the disc diffusion method (Bauer et al., 1966). The concentration of different antimicrobial discs was tetracycline (30 mg), co-trimoxazole (25 mg), cefuroxime (30 mg), cloxacillin (1 mg), cephaloxime (30 mg) and lincomycin (2 mg). The susceptibility was interpreted as sensitive, intermediate and resistant according to the zone size interpretation chart supplied by the manufacturer (HiMedia Laboratories, Bombay). Results and Discussion Characterization of 220 isolates of camel skin fungi revealed Candida albicans (14), Alternaria alternata (13), Epidermophyton floccosum (8), Scopulariopsis brevicaulis (10), Microsporum audouinii (2), M. canis (12), M. nanum (18), M. ferrugineum (5), Trichophyton verrucosum (15), T. mentagrophytes (3), T. schoenleinii (1), T. equinum (5), T. concentricum (2), T. tonsurans (2), T. violaceum (2), T. soudanense (6), T. rubrum (3), Sporothrix schenckii (1), Basidiobolus ranarum (3), Coccidioides immitis/posadasii (4), Penicillium marneffei (3), Penicillium spp.(4), Curvularia lunata (2), Exserohilum rostratum (1), Absidia corymbifera (5), Rhizopus oryzae (5), Aspergillus flavus (10), A. fumigatus (6), A. niger (12), A. terreus (13), A. versicolor (4), Aspergillus spp. (24) and Paracoccidioides spp. (2). Characterization of eight fungal isolates from cases of diarrhoea revealed R. oryzae (3); A. corymbifera (2) and Aspergillus spp. (3). Four blood sera samples from field cases with history of abortion were screened for brucellosis by Indirect ELISA at PDADMAS, Bangalore. One sample was found positive for antibrucella antibody titres. W hereas representative seven samples from pregnant animals of centre’s herd were found negative for antibrucella antibody titres. Skin infections were observed more in the winter season preceding rainy season and were more prevalent in very young calves (15 years), active grower (1-5yrs) and adults (Table.2 and 3). Actinobacillosis was more prevalent in young calves and the incidence decreased with the advancement of age of the animal (Table. 3). Mastitis was more prevalent in the peak yielders and incidence of abortion increased with the age of the animal (Table. 4). In a herd of 47 camels, 14 died after a heavy rain fall (about 150 mm) in the month of November, 2010 in the Pali

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district of Rajasthan. Of these, the 12 animals were sick with the signs and symptoms of initial constipation for 2-3 days followed by watery diarrhoea for next 2-3 days, then development of oedematous swelling on the abdomen, thighs and brisket region accompanied by colic leading to death within 2-3 days of onset of oedema. Other clinical signs observed were lachrymation, depression, partial anorexia and fever (103105 0F). Succession of oedematous swelling with syringe revealed a straw-coloured serous fluid. Cultural examination of blood and odematous fluid samples from sick animals, led to isolation of Pasturella spp. (Table 1). In considering overall antibiotic sensitivity 98.15 per cent of the bacterial isolates were found sensitive to tetracycline followed by Co-trimoxazole (78.20%), Cefuroxime (58.97%). Cephaloxime (53.85%), lincomycin and Cloxacillin (47.44% each). Tetracycline is still the drug of choice for treatment of various bacterial infections in camels (Table. 5). Amongst skin infections, fungal infections were more prevalent than the bacterial infections. Camel calves had higher infection rates than adult camels, which could be related to the development of a stronger immunity in older animals due to the multiplicity of exposure to the fungus rather than an intrinsic role of age (Descamps et al., 2003 and Moriello et al., 2003). Higher incidence of dermatophytosis was observed in the winter preceding rainy season. Gitao et al. (1998) reported higher infection rates of camel dermatophilosis from Kenya, Sudan and Saudi Arabia in the wet season as compared to the dry season. Identification and characterization of camel skin fungi along with type of the lesions has been described in our earlier reports (Tuteja et al., 2010a; Tuteja et al., 2010b; Tuteja et al., 2012; Tuteja et al., 2013a; Tuteja et al., 2013b and Tuteja et al., 2014). Mastitis was mainly caused by S. aureus a major mastitis pathogen and was more prevalent in peak yielders between 612 years of age (Table 1, 4). Predominance of staphylococci in camel mastitis has also been reported earlier (Tuteja et al., 2003; Bhatt et al., 2004 and Abdurahman, 2006). Camels are not known to be primary hosts for the Brucella organisms, but they are susceptible to pathogenic Brucella melitensis and B. abortus and the infection rate depends upon the infection rate in primary hosts animals in contact with them (Agab et al., 1994). Difficulties can arise in diagnosis of camel brucellosis, especially as this disease provokes only few clinical signs in contrast to its clinical course in cattle (Gwida et al., 2012). Epidemiological evidence shows that in India, brucellosis is present in different species of mammalian farm animals including cattle, goats, buffalo, yaks, camel, horses and pigs (Renukaradhya et al., 2002; Isloor et al., 1998; Mehra et al., 2000; Chakraborty et al., 2000; Kumar et al., 1997; Mrunalini et al., 2000 and Singh et al., 2000). The occupational exposure due to brucellosis is high in countries where herding of animals is still traditional and unscientific (Straten et al., 1997). Human infection from camels is known to occur mostly through the consumption of raw milk (Kiel and Khan, 1989). Malta fever due to B. melitensis was diagnosed in 30% of the camel handlers and milkers on large camel farm in Riyadh, Saudia Arabia. The abortion rate in the farm reaching 12% and B. melitensis biovars 1, 2 and 3 were isolated from aborted camel fetuses (Radwan et al., 1995). Given the 29


June 2015

Table 1: Incidence of various ailments along with causative agents S.N Ailments and no. of Type and . cases no. of samples 1

Skin infection 409

2

Mastitis 46

3

Diarrhoea/Enteritis 33

4

Abortion 23

5

Actinobacillosis 16 Pasturellosis 12

6

Bacterial Isolates

Skin scrapings/ Pus swabs 282 Milk samples 35 Faecal swabs 20 Serum samples 18 Pus swabs 3 Blood/ odematous fluid 12

127

Bacteria identified No. of S. S. Coryne- Pseudo- Strepto Bacillus E. Pasturella fungal isolates aureus epidermi bacterium monas coccus spp. coli spp. dis spp. spp. spp. 39 29 47 1 11 220

35

25

1

6

-

3

-

-

-

15

-

1

10

-

-

3

1

-

8

+1/4 *

-

-

-

-

-

-

-

-

-

**

-

-

-

-

-

-

-

-

-

12

-

-

-

-

-

-

-

12

-

*Positive for antibrucella antibody titres. ** + ve by direct smear examination for actinobacillus.

Table 2: Prevalence of infectious diseases along with seasonal variations

S. N.

1 2 3 4 5 6

Ailment

Total no. of animals examined Skin infection History of abortion Mastitis Actinobacillosis Enteritis/ Diarrhoea Pasturellosis cases

Total Overall no percentage of cases 2248 100

Summer Mar-June No of % cases age 364

Rainy July -Oct No of % age cases 884

Winter Nov- Feb No of % age cases 1000

409 23 46 17 34

18.19 1.02 2.05 0.76 1.51

44 3 10 4 0

12.09 0.82 2.75 1.10 0

131 10 7 9 21

14.82 1.13 0.79 1.02 2.37

234 10 29 4 13

23.40 1.00 2.90 0.40 1.30

12

0.53

0

0

0

0

12

1.20

Table 3: Prevalence of skin infections and actinobacillosis with age of the animals.

S. N. Age and stage of the animal 1

2 3 4 5

< 1 year (Immunologically weak) 1-5 year ( Active grower) 5-10 year (Adult) 10-15 year (Adult) >15 year (Aged)

Total no of animals examined 187

No. of the animals having skin infections 76

%

No of the % animals having actinobacillosis 3 1.60

40.64

608

119

19.57

7

1.15

916 420 117

131 62 21

14.30 14.76 17.95

6 1 0

0.65 0.23 0.00 30


Fig.1: Mature open abscess of actinobacillosis

Fig.2: Right quarter showing clinical mastitis

potentially huge economic and medical impact, a control policy could be cost-effective (Roth et al., 2003). India already has a policy for the control of brucellosis in dairy cattle (Renukaradhya et al., 2002). Prakash et al. (2012) concluded and recommended that as climatic conditions of Western Rajasthan mimics with Middle East where Brucellosis is prevalent, in clinical practice brucellosis should be kept in differential diagnosis in management of pyrexia of unknown origin and all preventive measure should be used for prevention of this zoonotic disease. A safe and effective vaccine in human is not yet available. Prevention is dependent upon increasing public awareness through health education programmes and safe livestock practices. Active co-operation between health and veterinary services should be promoted. Actinobacillus lignieresii is responsible for the wooden tongue disease characterised by the presence of granulomas with pus containing small, hard yellow to white granules. A. lignieresii affects mostly cattle and sheep but the disease is also known to occur in horses, pigs, dogs and chickens. In sheep, multiple purulent granules occur in the skin of the face, lips, nose, jaw and neck with regional lymph nodes usually being involved. The disease can be strongly suspected on

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Fig. 3: Lesions of skin infection

clinical grounds. Actinobacillus may be present in several different forms, depending on the specific agent and the host (Kahn and Line, 2010). In camels lesions were observed on the head and neck portion only as has been reported in case of sheep. Lesions were characterized microscopically by scattered pyogranulomas containing eosinophilic, club-like colonies surrounding gram-negative bacterial rods. The lesions were compatible with diagnosis of actinobacillosis. Purohit et al. (1988) suspected cases of cutaneous actinobacillosis in camels on the basis of lesions. Daneji et al. (1996) reported A. lignieresi infection in camels from Nigeria. Diarrhoea was commonly observed in the rainy season (2.37 %) followed by winter season (1.30%). No case of diarrhoea was observed during summer season. Out of the 20 diarrhoeic faecal samples examined 10 were infected with the Gram positive, club shaped Corynebacterium spp. Diarrhoea mixed with blood was the common finding in the cases infected with Corynebacterium spp. (Table 1). Earlier there does not appear to be such report of diarrhoea in camels due to Corynebacterium spp. But bloody diarrhoea in camels has been reported in the cases of pasturellosis (Wernery and Kaaden, 2002). Sudden climate change in the dry lands affects the desert livestock species adversely. Varible but some corresponding symptoms in pasturellosis has been reported by various workers (Wernery and Kaaden., 2002). Considering the overall antibacterial sensitivity (table 5); 98.15 per cent of the isolates were found sensitive to tetracycline followed by co-trimoxazole (78.20%), cefuroxime (58.97%). cephaloxime (53.85%), lincomycin and cloxacillin (47.44% each). Tetracycline is still the drug of choice for treatment of various bacterial infections in camels. High sensitivity observed with tetracycline in this study is in agreement with our previous study (Tuteja et al., 2003) and has been reported by Hawari and Hassawi (2008) also. Acknowledgements Authors are thankful to the Director, National Research Centre on Camel, Bikaner, Rajasthan, India for providing necessary facilities to carry out the present study.

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Table 4: Incidence of cases of clinical mastitis and abortions as per age and stage of the animals.

S. N.

1 2 3

Age of the animal up to 6 years 6-12 yeras > 12 years

No of Stage of females in the the age animal group 269 Fresh calvers 551 Peak yielders 246 Aged lactators

No of cases of clinical mastitis 9 27 10

%

Stage of the No of cases animal of abortions

3.35 Fresh pregnancies 4.9 Mid pregnancies 4.06 Aged pregnancies

% +ve

3

1.11

13

2.35

7

2.85

Table 5: Antibiotic sensitivity of bacterial isolates.

Organisms and number of isolates S. S. Coryne- Bacillus PseudoAntibiotic Sensitivity aureus epidermidis bacterium spp. monas (20) (28) spp. (18) (10) spp. (2) S 19 28 17 10 1 Tetracycline I 1 1 R 1 S 12 26 14 8 1 CoI 2 2 trimoxazole R 8 2 2 1 S 6 14 9 7 1 Cloxacillin I R 14 14 9 3 1 S 5 12 11 8 1 Lincomycin I R 15 16 7 2 1 S 7 17 14 7 1 Cefuroxime I 2 6 R 11 5 4 3 1 S 10 16 11 4 1 Cephaloxime I 8 12 5 1 R 2 7 1 References Abbas, B. and Agab, H. (2002) Prev. Vet. Med. 55: 47-56. Abbas, B. and Tilley, P. (1990) Nomadic Peoples. 29: 77-86. Abbas, B. et al. (1993) Sudan J. Vet. Sci. Anim. Hus. 32: 31-41. Abdurahman, O.A.Sh. (2006) Livest. Res. Rural Dev. 18: 1-9. Agab, H. (1993) Epidemiology of Camel Diseases in Eastern Sudan with Emphasis on Brucellosis. M.V.Sc. Thesis. University of Khartoum. pp. 172. Agab, H. et al. (1994) Rev. Elev. Med. Vet. Pays. Trop. 47: 361-363. Bauer, A.W. et al. (1966) Amer. J. Clin.Path. 45: 493-496. Bhatt, L. et al. (2004) Vet. Pract. 5: 61-65. Brown, R.W. et al. (1981) Microbiological procedures for use in the diagnosis of bovine mastitis. 2nd ed. National Mastitis Council Inc., 30F Street, N.W. Washington D.C. Chakraborty, M. et al. (2000) Indian Vet. J. 77: 924-925. Dalling, T. et al. (1988) Diseases of camels. In: The International Encyclopedia of Veterinary Medicine. Edinburgh, U.K., W. Green and Son. 585. Daneji, A.I. et al. (1996) Trop. Anim. Health Prod. 28: 315-316. Descamps, F. et al. (2003) FEMS Immunol. Med. Microbiol. 38: 2934. El-Gayoum, S.E.A. (1986) Study on the mechanism of resistance to camel diseases. Thesis, Gottingen 22.

Total (78) 75 2 1 61 4 13 37 0 41 37 0 41 46 8 24 42 26 10

Per cent 96.15 2.57 1.28 78.20 5.13 16.67 47.44 0 52.56 47.44 0 52.56 58.97 10.26 30.77 53.85 33.33 12.82

Gitao, C.G. et al. (1998) Proc. 3rd Annual meeting for animal production under arid conditions. United Arab Emirates University.2: 93-107. Gwida, M. et al. (2012) Res.Vet. Sci. 92:351-355. Hawari, A.D. and Hassawi, D.S. (2008) J. Biol. Sci. 8: 958-961. Isloor, S. et al. (1998) Rev. Sci. Tech. 17: 781-785. Kachhawaha, S. et al. (2012) Vet. Pract. 13: 218-222. Kahn, M.C. and Line, S. (2010) The Merck Veterinary Manual. 10th ed. Published by Merck & Co., Inc. Whitehouse Station, N.J., U.S.A. Kiel, F.W. and Khan, M.Y. (1989) Soc. Sci. Med. 29: 999-1001. Kumar, P. et al. (1997) Indian J. Anim. Sci. 67: 180-182. Margan, Ute. (1987) Vergleichende Untersuchungen zur Bedeutung der alternativen Komplementaktivierung bei Rindern und Kamelen. Thesis, Gottingen 33. Mehra, K.N. et al. (2000) Indian Vet. J. 77: 571-573. Moriello, K.A. et al. (2003) J. Feline Med. Surg. 5: 161-166. Mrunalini, N. et al. (2000) Indian Vet. J. 77: 932-935. Prakash, P. et al. (2012) Natl. J. Comm. Med. 3: 62. Purohit, N.R. et al. (1988) Aust. Vet. J. 65:31-32. Qureshi, S. et al. (2002) J. Camel Pract. Res. 9: 129 -134. Radwan, A.I. et al. (1995) Rev. Sci. Tech. 14:719-732. Renukaradhya, G.J. et al. (2002) Vet. Microbiol. 90: 183-195.

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