Patient 1

A 39-year-old male pest controller from Gladstone on a routine visit to his general practitioner in April 2001 inquired about the recent appearance of a red macule, 8 mm in diameter, on the medial aspect of his right ankle. No specific treatment was given. When he was seen again 18 days later, a painful, necrotic ulcer, about 12 mm in diameter, had developed at the original site of the red spot. An exudates was produced, although polymorphs were observed rarely, and no microorganisms were identified on Gram stain. A gram-negative organism later identified as Photorhabdus sp. was isolated in pure growth from the exudate. The patient began a 10-day course of oral cephalexin. When he was observed again 11 days later, he exhibited a persistent discharge with surrounding cellulitis. He was therefore prescribed a 10-day course of oral amoxycillin-clavulanate. Three weeks later, the ulcer appeared to be healing; after another 6 weeks, signs of infection had again developed. A gram-negative organism was isolated from the exudate but was not formally identified.

The patient was prescribed an additional 7-day course of oral cephalexin. When he was observed 3 months later, the infection had resolved. In his recent work as a pest controller, he had been spraying chemical insecticides under houses and in foreign cargo ships. He had never used insect pathogenic nematodes as a biopesticide.

Patient 2

A 78-year-old man from the Queensland Gold Coast sought treatment in January 1999 with a 3-day history of a painful, swollen right foot. The patient had a history of polymyalgia rheumatica for which he was taking prednisone, 8 mg daily. In January 1999, after working barefoot in the garden, the man noted intense pain in his right forefoot and a very small amount of bloody discharge from the web space between his fourth and fifth toes.

The next day he was seen by his general practitioner who treated him with oral dicloxacillin. Two days later he was admitted to the hospital with increasingly severe pain with extensive redness and swelling extending to his right knee. He was noted to be afebrile with a mild neutrophil leukocytosis. He was started on a regimen of intravenous dicloxacillin and gentamicin.

Surgical debridement of the right foot was required on three occasions during the first 8 days of his admission. Pus was collected for culture on two of these occasions, and tissue was obtained during the third. An organism identified as Photorhabdus sp. was isolated in pure culture from each of these operative specimens. The same organism was also isolated, together with Staphylococcus aureus, from a superficial swab collected in the emergency department on presentation. No bacterial growth was obtained from blood cultures collected on admission.

The patient was treated with intravenous gentamicin for 2 weeks and ceftazidime for 1 week. He was discharged on a 6-week course of oral ciprofloxacin. The foot remained healed on follow-up 3 months later.

Photorhabdus spp. can be isolated and identified to genus level by using techniques available in most clinical bacteriology laboratories. A total of five isolates from the two patients described in the current report were examined in our laboratories with standard techniques (one from patient 1 and four from patient 2). The phenotypic characteristics that the isolates displayed were typical of the genus.

Figure 1

Figure 1. Photorhabdus isolate from patient 2, growing on tryptic soy agar containing 5% sheep blood, after 48 hours’ incubation at 35°C. Arrows indicate “swarming.” The colonies could be seen to glow faintly...

Figure 2

Figure 2. Photorhabdus isolate from patient 2 after 5 days growth at room temperature on sheep blood agar. Arrows indicate the characteristic thin line of “annular” hemolysis surrounding the colonies.

Colonies were formed after 24–48 hours on tryptic soy agar containing either 5% sheep or horse blood (bioMérieux, Baulkham Hills, Australia) at both 35°C and at room temperature, with a tendency to “swarm” (Figure 1). The isolates also grew on MacConkey agar. On sheep and horse blood agar, a thin line of annular hemolysis was observed 4–12 mm from the colony edge. The hemolysis was more apparent when the isolates were incubated at room temperature (Figure 2). The organisms were motile, gram-negative, rod-shaped bacteria. They were facultatively anaerobic, oxidase negative, and strongly catalase positive. Other biochemical reactions were as described previously (1).

The defining characteristic was the presence of faint luminescence, which could be clearly seen with the naked eye when the colonies were examined under conditions of total darkness. It was critical to this examination that the observer’s eyes be allowed to adjust to the darkness for 10 minutes.

Two commercially available automated bacterial identification systems were used in our laboratories: MicroScan Walkaway (Dade Behring Inc., MicroScan Division, West Sacramento, CA) and bioMerieux Vitek (bioMérieux; Hazelwood, MO). Photorhabdus spp. do not currently appear on the databases of either of these systems, which leads to misidentification (Table 1).

Photorhabdus spp. have been shown to form a heterogeneous group based on DNA-DNA hybridization studies, 16S rDNA sequencing and polymerase chain reaction ribotyping (2). A polyphasic approach is now applied to classifying isolates within the genus, dividing it into three species and several subspecies. The American clinical isolates described by Farmer et al. (6) belong to a new species, Photorhabdus asymbiotica (2). A specific epithet has not yet been assigned to the Australian clinical isolates but they also may form a new species within the genus (7).

Antimicrobial sensitivity was assessed by using broth microdilution. The isolates were sensitive to a broad range of antimicrobial agents with activity against gram-negative bacteria including ciprofloxacin, gentamicin, tetracycline, ceftriaxone, and amoxycillin-clavulanate. Isolates from both patients were resistant to cephalothin and ampicillin.