Fatal Actinobacillosis in a Dairy Buffalo

ACTA VET. BRNO 2006, 75: 247–250 Fatal Actinobacillosis in a Dairy Buffalo G. MUHAMMAD, M. SAQIB, M. ATHAR Department of Veterinary Clinical Medicine and Surgery, Faculty of Veterinary Science, University of Agriculture, Faisalabad, Pakistan Received August 23, 2005 Accepted March 16, 2006 Abstract Muhammad G., M. Saqib, M. Athar: Fatal Actinobacillosis in a Dairy Buffalo. Acta Vet. Brno 2006, 75: 247-250. The present report describes a case of fatal actinobacillosis in a dairy buffalo (Bubalus bubalis). Clinical signs included a large multilobular, firm abscess located caudo-ventral to the right ear base, submandiblar and prescapular lymphadenopathy, fever and dyspnoea. Haemogram revealed an accelerated erythrocyte sedimentation rate and erythropenia. Provisional diagnosis of actinobacillosis was confirmed by the demonstration of a Gram-negative pleomorphic organism in pus, microscopic examination of a compression preparation of a sulphur granule as well as growth of the organism on blood agar. Intraperitoneal inoculation of pus in guinea pigs was fatal at around 48 hours post-injection. The LD50 of the Actinobacillus lignieresii isolate in mice was 104 colony forming unit per milliliter of Hank’s balanced salt solution. The animal died 12 hours after the initiation of surgical drainage of the abscess and administration of iodide, antibiotics and steroids. Necropsy findings included interlobular pneumonia, pulmonary abscessation and mediastinal lympadenopathy that were ascribed to metastatic spread of the organisms to the lungs which probably led to the death of the animal. This case report appears to be the first documentation of fatal actinobacillosis in the dairy buffalo. Actinobacillosis, Actinobacillus lignieresii, Bubalus bubalis, dairy buffalo Actinobacillosis is an infectious, chronic, generally non-fatal disease caused by Actinobacillus lignieresii. In cattle, it typically involves the tongue and hence is also known as “the Wooden Tongue”. The involvement of other organs is considered to be atypical (Mohanty and Singh 1970; Rebhun et al. 1988; Aslani et al. 1995; Holzhauer and Roumen 2002). The etiologic agent is found normally in the oropharynx and rumen of cattle and sheep (Smith 1990; Songer and Post 2005). The organism is an opportunistic pathogen and it causes chronic pyogranulomatous lesions of the soft tissues of head and neck regions in cows, buffalo, sheep, goats, and horses (Misra and Angelo 1981; Fubini and Campbell 1983; Baum et al. 1984; Kumar and Parihar 1998; Hirsh et al. 2004; Songer and Post 2005). Clinically, the lesions appear as nodules, multiple abscesses, ulcers or draining fistulae. Spontaneous actinobacillosis in the buffalo (Bubalus bubalis) is reportedly non-fatal (Akhtar and Cheema 1964; Ahmad et al. 1986). The present report describes a case of atypical, fatal actinobacillosis in a dairy buffalo. Case Report An 8-year old, approx. 500 kg Nili-Ravi dairy buffalo in the 4th month of her 5th lactation was presented to the Veterinary Medical Teaching Hospital, University of Agriculture, Faisalabad, Pakistan, for the treatment of a large, lobulated abscess at the base of the right ear and additional swellings cranial to both the right and left shoulders. Anamnesis indicated that the condition began five months previously as a small, walnut-size swelling below the base of the right ear. The swelling gradually enlarged over this period and became lobulated. One of lobules of the abscess located at the ear base broke open about 3 weeks prior to presentation. The animal was treated with a longacting preparation of oxytetracycline for two days before presentation. The animal was also anorexic for the last couple of days. Address for correspondence: Dr. M. Saqib, Department of Veterinary Clinical Medicine & Surgery University of Agriculture, Faisalabad 38040, PAKISTAN Phone: +92(41) 9200161/3110 Fax: +92(41) 9200204 Email: [email protected] http://www.vfu.cz/acta-vet/actavet.htm 248 On clinical examination, the buffalo exhibited fever (103 °F; 39.4 °C), mild dyspnoea and halitosis. The most noticeable sign was the presence of a large (33 × 18 cm), warm and firm, lobulated abscess located caudo-ventral to the right ear. This abscess had the contour of four tennis ball-sized lobules as well as a walnut-sized one. The posterior-most lobule located just adjacent to the ramus of the mandible had a small deep sinus that was discharging a mucoid, greyish-white, slightly putrid, non-gritty pus. Right and left prescapular lymph nodes were grossly enlarged to the size of a muskmelon (approximately 18 × 15 cm). The submandibular lymph nodes were also extensively enlarged. Significant haematological alterations included an accelerated erythrocyte sedimentation rate (120 mm after first hour) and erythropaenia (4.1 million µl-1). Using a sterile disposable syringe, about 10 ml of greyish-white mucoid pus was aspirated from the dorsal lobule of the abscess located directly below the right ear. Microscopic examination of Gram-stained smears of the pus revealed a Gram-negative organism in isolated coccoid, filamentous and club forms. Culture of this pus on blood agar plates yielded small (1 to 2 mm), raised, greyishwhite, non-haemolytic colonies within 24 hours. Gram-stained smears of these colonies revealed small Gramnegative rods. Intraperitoneal (IP) inoculation of 1 ml of pus diluted with an equal volume of saline into two guinea pigs (in accordance with the requirements of laboratory animal use in biomedical research as defined by The Animal Ethics Committee, University of Agriculture, Faisalabad) led to distension of the peritoneal cavity with a turbid serous fluid as well as the guinea pigs’ deaths at around 48 hours post-injection. After the aspiration of pus with a syringe, the dorsal mature pocket of the abscess below the right ear was surgically lanced and about 50 ml of pus was collected in a Petri dish. Careful exploration of this pus revealed only a single whitish-yellow pinhead-sized sulphur granule. After washing in sterile water, this granule was crushed between two microscope slides and Gram stained. Microscopic examination of the stained smears showed Gramnegative bacilli, cocci and short filaments. The combination of clinical signs, cultural and microscopic examination of the pus and a sulphur granule provided the basis for diagnosis of actinobacillosis in the subject of the present report. The upper-most matured pocket of the lobulated abscess below the right ear was surgically lanced, drained and flushed with a 1% solution of povidone-iodine Pyodine, Brooks Labs., Pakistan). Twelve grams of potassium iodide was dissolved in water and the buffalo was drenched with it. Twenty millilitres of trimethoprimsulphadiazine (Inj. Tribrissen 48%, Glaxo-Wellcome-Beechem, Pakistan) and 10 ml of prednisolonedexamethasone (Inj. Pre-Dexa, Shinil, Korea) were administered intramuscularly. The buffalo died 12 hours after the institution of therapy. Post mortem examination revealed blood-tinged fluid in the thoracic cavity and bilaterally congested lungs with almond-sized abscesses in their anterior lobes. Mediastinal lymph nodes were moderately enlarged. The material aspirated from the lungs abscesses and mediastinal lymph nodes yielded non-haemolytic, small, greyish white, raised colonies after 48 hours of culture on blood agar. The organism was catalase- and oxidase-positive. It fermented fructose, glucose, galactose, maltose, mannose, sucrose, and xylose without gas production. Methyl red, Voges-Proskauer and indole tests were negative. Four different dilutions (104 106, 10 8, 1010 colony forming unit (cfu) per millilitre of Hank’s balanced salt solution) of Actinobacillus lignieresii isolate were injected IP into sixteen mice; four mice per dilution. Four additional mice injected IP with 1 ml of Hank’s balanced salt solution alone, served as controls. The mice weighed 20-25 g. The injections were made into the left lower abdominal quadrant with a 25-gauge needle. The most dilute inoculum (104 cfu/ml) culminated in the death of two of the four mice at 24 and 27 hours post inoculation. The remaining three concentrations (106, 108 and 1010 cfu/ml) led to the death of all the inoculated mice between 18-21, 9-11.5 and 9-11 hours post inoculation, respectively, whereas all mice in the control group survived. The organism was re-isolated from all mice inoculated with the bacterial suspension. Discussion As far as could be ascertained from the literature, the present report appears to be the first record of fatal spontaneous actinobacillosis in the dairy buffalo. In reported outbreaks of actinobacillosis in the buffalo (Akhtar and Cheema 1964; Ahmad et al. 1986), the case fatality rate was zero. Cheema (1965), however, documented that experimental inoculation of Actinobacillus lignieresii isolated from the buffalo was fatal in buffalo calves within seven months but not in bovine calves. Respiratory difficulty observed in the subject of the present report may be ascribed to metastatic extension of the disease to the lungs (Cheema 1965; Gill and Balwant-Singh 1977; Carter and Chengappa 1991). Similarly, extensive swelling of submandibular lymph nodes as well as right and left pre-scapular lymph nodes may be attributed to metastatic spread and localization in these glands (Shastry 1983; Aslani et al. 1995; Radostits et al. 2000). Fatality of guinea pigs after intraperitoneal inoculation concurs with the observation of Cheema (1965). The biochemical characteristics of the organism isolated from pus and characterized as A. lignieresii were consistent with those described by Lentsch and Wagner (1980). 249 Actinobacillosis in cattle typically involves the tongue, and the involvement of other organs is considered to be atypical. The disease process in the buffalo of the present report did not affect the tongue. Apart from a solitary report of lingual actinobacillosis in a buffalo (Misra and Angelo 1981), all other reports on actinobacillosis in the buffalo describing sporadic cases (e.g. Panchbhai and Deshpande 1985) or outbreaks of this disease (Akhtar and Cheema 1964; Ahmad et al. 1986) documented only cutaneous involvement without tongue involvement. It may be that contrary to bovine actinobacillosis, bubaline actinobacillosis typically involves the skin and not the tongue. Additional clinical research is clearly warranted to verify or refute this tentative conclusion. Fatální aktinobacilóza u buvola domácího Práce popisuje pfiípad fatální aktinobacilózy u buvola domácího (Bubalus bubalis). Klinick˘m nálezem byl velk˘ multilobulární absces, kaudoventrálnû od báze pravého ucha, lymfadenopatie (submandibulární a preskapulární mízní uzliny), horeãka a dyspnoe. Hematologické vy‰etfiení ukázalo zv˘‰enou a zrychlenou sedimentaci a erytropenii. PfiedbûÏná diagnóza aktinobacilózy byla potvrzena nálezem gram-negativních pleomorfních organismÛ v hnisu, mikroskopického vy‰etfiení kompresního preparátu a rÛstu mikroorganismÛ na krevním agaru. Intraperitoneální aplikace hnisu byla pro morãata fatální kolem 48 hod po aplikaci. Poloviãní letální dávka, LD50 izolátu Actinobacillus lignieresii byla u my‰í 104 CFU.ml-1 (colony forming unit) Hankovy soli. Buvol uhynul 12 hodin po zahájení chirurgické drenáÏe abscesu a aplikaci iodidu, antibiotik a steroidÛ. Pfii pitvû byla zji‰tûna interlobulární pneumonie, abscedace v plicích a mediastinální lymfadenopatie, která byla pfiipisována metastatickému ‰ífiení mikroorganismu do plic, coÏ pravdûpodobnû vedlo k uhynutí zvífiete. Tento pfiípad je pravdûpodobnû první popsaná fatální aktinobacilóza u buvola domácího. Acknowledgements Authors thank Dr. Gila A. Sutton, Koret School of Veterinary Medicine, Faculty of Agricultural, Food and Environmental Quality Sciences, The Hebrew University of Jerusalem, Israel for reviewing this manuscript. References AHMAD A, MAJEED MA, JABBAR MA 1986: Blood picture of naturally occurring actinobacillosis in male buffalo calves. Pakistan Veterinary J 6: 80-83 AKHTAR AS, CHEEMA RA 1964: Actinobacillosis in buffaloes. Proceedings of 16th Pakistan Science Conference on Medicine and Veterinary Science. G-24 ASLANI MR, KHODAKARAM A, REZAKHANI A 1995: An atypical case of actinobacillosis in a cow. J Vet Med (Series A) 42 : 485-488 BAUM KH, SHIN SJ, REBHUN WC, PATTEN VH 1984: Isolation of Actinobacillus lignieresii from enlarged tongue of a horse. J Am Vet Med Assoc 185:792-793 CARTER GR, CHENGAPPA MM 1991: Essentials of Veterinary Bacteriology and Mycology. Lea & Febiger, Philadelphia, pp. 179-182 CHEEMA RA 1965: Systematic bacteriology and transmission studies of strains of Actinobacillus lignieresii isolated from buffaloes of Lyallpur. M.Sc. Thesis, West Pakistan Agricultural University, Lyallpur, Pakistan FUBINI SL, CAMPBELL SG 1983: External lumps on sheep and goats. Veterinary Clinics of North America: Large Animal Practice 5: 457-476 GILL, BS, BALWANT-SINGH 1977: Pulmonary actinobacillosis and actinomycosis in Indian buffaloes. Orissa Vet J 11: 104-109 HIRSH DS, MACLACHLAN NJ, WALKER RL 2004: Veterinary Microbiology, 2nd ed., Blackwell Publishing Professional, Ames, Iowa, pp. 91-94 HOLZHAUER M, ROUMEN TP 2002: Atypical actinobacillosis in a dairy replacement herd. Vet Rec 151: 276 KUMAR GS, PARIHAR NS 1998: Respiratory ailments in Indian buffaloes. Buffalo J 14: 109-114 LENTSCH RH, WAGNER JE 1980: Isolation of Actinobacillus lignieresii and Actinobacillus equuli from laboratory rodents. J Clin Microbiol 12: 350-353 MISRA SS, ANGELO SJ 1981: Treatment of lingual actinobacillosis in a buffalo. Indian Vet J 58: 413- 414 250 MOHANTY GC, SINGH CM 1970: Actinobacillotic nephritis in a male buffalo: a case report. Indian Vet J 47: 938-941 PANCHBHAI VS, DESHPANDE BB 1985: A clinical case of cutaneous actinobacilosis in a buffalo. Indian Vet J 62: 525-526 RADOSTITS OM, GAY CC, BLOOD DC, HINCHCLIFF KW 2000: Veterinary Medicine, W.B. SAUNDERS, Philadelphia, pp. 736- 737 REBHUN WC, KING JM, HILLMAN RB 1988: Atypical actinobacillosis granulomas in cattle. Cornell Vet 78: 125- 130 SHASTRY GA 1983: Veterinary Pathology. CBS Publishers and Distributors, New Delhi, India, pp. 49-51 SMITH BP 1990: Large Animal Internal Medicine. The C.V. Mosby Co. Philadelphia, pp. 719-721 SONGER JG, POST KW 2005: Veterinary Microbiology - Bacterial and Fungal Agents of Animal Disease, ELSEVIER SAUNDERS, St. Louis, Missouri, pp. 174-180