Germán B. Vigo

OUTBREAKS OF AVIAN CHOLERA IN HOPE BAY, ANTARCTICA

During austral summers 1999–2000 and 2000–01, two outbreaks of avian cholera occurred in the Hope Bay area (63°24′S, 56°59′W), located on the tip of the Antarctic Peninsula. Eighty-six dead birds were found: five kelp gulls (Larus dominicanus), 36 skuas (Stercorarius sp.), and 45 Adelie penguins (Pygoscelis adeliae). The carcasses were studied using clinical, pathological, and microbiological criteria. Water samples from ponds where birds were settled and samples from 90 healthy birds also were analyzed during the second outbreak. Pasteurella multocida isolates were identified by biochemical tests, capsular type, somatic serotype, and susceptibility to nine antibiotics. Molecular subtyping was performed by ApaI and SmaI pulsed-field gel electrophoresis (PFGE) and enterobacterial repetitive intergenic consensus (ERIC-PCR). In February 2000, mortality in skuas was 16% and 2% in kelp gulls. In the 2000–01 breeding season, mortality in south polar skuas was 47%, 24% in brown skuas, 1.4% in kelp gulls, and 0.01% in Adelie penguins. All birds had lesions of avian cholera. In kelp gulls the presentation was chronic, whereas skuas and penguins suffered subacute and acute disease, respectively. Fifty-five isolates recovered from dead birds and one from water were identified as P. multocida gallicida, type A:1. The strains presented a unique molecular pattern by PFGE and ERIC-PCR. A possible hypothesis to explain the origin of the outbreaks was that nonbreeder kelp gulls carried P. multocida gallicida to Hope Bay, and avian cholera was transmitted through water to skuas and penguins. This study reports avian cholera in new bird species, their potential role in the transmission of the disease, and the different responses of these species to the disease.

Detection of virulent Rhodococcus equi in tracheal aspirate samples by polymerase chain reaction for rapid diagnosis of R. equi pneumonia in foals

Polymerase chain reaction (PCR)-based assays were developed to detect virulent Rhodococcus equi in transtracheal aspirate samples from sick foals showing respiratory signs. An oligonucleotide primer pair from the sequence of the virulence-associated 15- to 17-kDa antigen gene of the virulence plasmid in virulent R. equi was used to amplify a 564 bp region by PCR, and the result was confirmed by Southern blot hybridization. No positive reaction was seen in DNA from 13 different microorganisms typically found in the respiratory tract. In tracheal aspirates seeded with virulent R. equi, a visible band could detect 10 to 10(2) bacteria per PCR assay (10(3) to 10(4)/ml of the aspirate). Virulent R. equi was demonstrated in 31 of 42 transtracheal aspirates by culture and colony blot analysis, whereas a positive PCR result was observed in only 12 of the 31 culture positive samples. To prevent false-negative results, two methods were developed: a nested PCR and a PCR in combination with enrichment cultures of aspirates in the selective medium to increase the number of bacteria to 10(4)/ml or more. All of the PCR-negative and culture-positive samples were positive by the two methods. These results indicated that PCR-based assays provide a specific and sensitive means to detect virulent R. equi in tracheal aspirates of foals, and they are more rapid than the routine culture procedures for the diagnosis of R. equi pneumonia in foals.

Avian Cholera in a Southern Giant Petrel (Macronectes giganteus) from Antarctica

A southern giant petrel (Macronectes giganteus) was found dead at Potter Peninsula, King George Island, South Shetland, Antarctica. The adult male was discovered approximately 48 hr after death. Macroscopic and microscopic lesions were compatible with avian cholera and the bacterium Pasteurella multocida subsp. gallicida, serotype A1 was isolated from lung, heart, liver, pericardial sac, and air sacs. In addition, Escherichia coli was isolated from pericardial sac and air sacs. This is the first known report of avian cholera in a southern giant petrel in Antarctica.

Salmonella enterica Subclinical Infection: Bacteriological, Serological, Pulsed-Field Gel Electrophoresis, and Antimicrobial Resistance Profiles—Longitudinal Study in a Three-Site Farrow-to-Finish Farm

The aim of this surveillance was to study both Salmonella spp. shedding patterns and the time course of serological response in farrow-to-finish reared pigs from a subclinically infected farm. Antimicrobial resistance profile, molecular subtyping, and the relationship among the isolates were determined by pulsed-field gel electrophoresis (PFGE). A farrow-to-finish farm of 6000 sows, with a history of Salmonella Typhimurium septicemia, was selected. A longitudinal bacteriological and serological study was conducted in 25 sows before farrowing (M/S1) and in 50 offspring at 21 (M/S2), 35 (M/S3), 65 (M/S4), 86 (M/S5), 128 (M/S6), and 165 (M/S7) days of age. Serum antibodies were tested using Herdcheck((R)) Swine Salmonella antibody test kit (Idexx Laboratories, ME). Bacteria were isolated from pooled fecal samples. Suspected isolates were confirmed by conventional biochemical assays, and those identified as Salmonella spp. were serotyped. A variation between seropositive percentages and positive fecal samples was observed. Serologically positive pigs decreased from S1 to S4, and subsequently increased from S4 to S7. The percentages of fecal positive culture increased from M1 to M3, and then declined in M4, increased in M5, and were negative in M6 and M7. In the study three serovars, Salmonella 3,10:e,h:-, Salmonella Muenster, and Salmonella Bovismorbificans, were identified with low pathogenicity for swine. Three multidrug resistance strains (one belonged to Salmonella 3,10:e,h:- and two belonged to Salmonella Muenster) were found. PFGE results showed three different but closely related patterns among the 13 isolates of Salmonella Bovismorbificans, and two patterns for the three Salmonella Muenster and Salmonella 3,10:e,h:- isolates. This longitudinal study established critical points of Salmonella spp. infection in the farm and the production stages, where appropriate control measures must be taken. PFGE showed clonal relationships in each serovar. Antibiotic resistance profiles should be periodically included due to public health concerns.

Resistencia a los antimicrobianos en bacterias indicadoras y zoonóticas aisladas de animales domésticos en Argentina

Antimicrobial resistance profiles in indicator and zoonotic bacteria isolated from faeces of healthy animals without clinical signs of the following species: bovine, equine, ovine, porcine, layer hens, and canine, were studied. The chosen antimicrobials are frequently used in veterinary and human medicine. The agar diffusion was the method used. The obtained results of 240 Escherichia coli, 189 Enterococcus spp., 11 Campylobacter spp. and 2 Salmonella Gaminara (16:d:1,7) showed a greater percentage of resistance and multiresistance in intensive breeding animals, porcine and layer hens. The observed resistance to ampicillin, streptomycin, tetracycline and nalidixic acid in E. coli coincides with the antimicrobials most commonly used on animal farms, the same as tetracycline and erythromycin in Enterococcus spp. The strains of Salmonella Gaminara (16:d:1,7) were susceptible to the antimicrobials tested. In Campylobacter spp. the scarce number of isolates hindered an adequate interpretation of the results. Owing to the lack of data in our country on antimicrobial resistance in indicator and zoonotic bacteria in domestic animals, we consider that the obtained values could be used as a starting point for a future monitoring program.

Prevalence of Edwardsiella tarda in Antarctic wildlife

For many years, the Antarctic region has been isolated from human activity. However, there is little data available regarding endemic and exotic diseases. The purpose of this work was to determine the prevalence of Edwardsiella tarda in Antarctic wildlife, including birds, mammals and fish. During the summer of 2000 and 2002 in the Potter Peninsula, and during the summer of 2001 and 2003 in Hope Bay, a total of 1,805 faecal samples from Antarctic animals and 50 infertile eggs of Adelie penguins (Pygoscelis adeliae) were collected in order to isolate E. tarda. The classic Edwardsiella tarda was isolated from 281 (15.1%) of the 1,855 Antarctic wildlife samples. This is the first report of E. tarda isolation from southern giant petrels (Macronectes giganteus), brown skuas (Stercorarius lonnbergi), south polar skuas (Stercorarius maccormicki), kelp gulls (Larus dominicanus), greater sheathbills (Chionis albus), chinstrap penguins (Pygoscelis antarctica), eggs of Adelie penguins and Weddell seals (Leptonychotes weddelli). None of the evaluated animals showed clinical signs of disease. Our results suggest that E. tarda is a common bacterium amongst Antarctic birds and mammals.

Thelebolus microsporus Mycelial Mats in the Trachea of Wild Brown Skua (Catharacta antarctica lonnbergi) and South Polar Skua (C. maccormicki) Carcasses

Sixteen brown skuas (Catharacta antarctica lonnbergi) and seven South Polar skuas (C. maccormicki) were found dead near Boekella Lake, Hope Bay, Antarctica, in February 1997. Postmortem examination revealed conspicuous caseous, deep yellow fungal/mycelial mats or cores in the trachea of nine of 19 carcasses that were examined. These mycelial cores, highly suggestive of aspergillomas, completely occluded the tracheal lumen in four of these nine carcasses. Thelebolus microsporus, a psychrophilic ascomycetous fungus commonly isolated from skua dung and skua nesting material, was isolated in pure culture from these tracheal plugs. Awareness of pseudolesions resulting from Thelebolus microsporus profuse postmortem growth in the trachea of dead skuas will minimize potential confusion with aspergillosis when investigating causes of epornithics in Antarctica.

Isolation and characterization of Salmonella enterica from Antarctic wildlife

In recent years, the human presence in Antarctica has increased and as a consequence, the possibility of microorganisms’ introduction. The aims of this work were to determine the presence of Salmonella enterica in Antarctic seabirds and sea mammals, to characterize the isolates identified, and to determine the genetic relation of Antarctic S. enterica isolates among them and compare with isolates of human, animal, and food sources recovered in Argentina. During the summer 2000 and 2002 in Potter Peninsula, and during the summer 2001 and 2003 in Hope Bay, a total of 1,739 fecal samples from Antarctic animals were collected and analyzed. In summer 2000, S. Newport and S. Enteritidis were isolated from 8.9% of southern giant petrels (Macronectes giganteus). In summer 2003, S. Enteritidis was isolated from 1.5% of Adelie penguins (Pygoscelis adeliae), from 5.5% of skuas (Stercorarius sp.), from 5.4% of kelp gulls (Larus dominicanus), and from 5.6% of Weddell seals (Leptonychotes weddelli). All the isolates belonging to the same serovar showed indistinguishable genomic profiles by Pulse-Field Gel Electrophoresis (PFGE) with XbaI and BlnI restriction enzymes and by Random Amplified Polymorphic DNA (RAPD-PCR). In addition, these Antarctic strains were different from S. enterica isolates from different sources identified in Argentina during the same or close time periods.

Isolation of Salmonella Typhimurium from Dead Blue and Gold Macaws (Ara ararauna)

Abstract Two blue and gold macaw (Ara ararauna) chicks died of fatal salmonellosis in Buenos Aires Province, Argentina. The birds were histopathologically and microbiologically examined. Salmonella enterica subspecies enterica serovar Typhimurium was isolated from the liver, spleen, heart, lung, kidney, and intestine of both birds. All strains were susceptible to ampicillin, cephalothin, cefotaxime, enrofloxacin, nalidixic acid, gentamicin, streptomycin, chloramphenicol, fosfomycin, tetracycline, nitrofurantoin, and trimethoprim-sulfamethoxazole. The XbaI-PFGE profile of the Salmonella Typhimurium isolated from the two animals, which shared the same cage, was identical and showed a unique pattern compared with 301 isolates included in the PulseNet national database of Salmonella pulsed-field gel electrophoresis patterns. This is the first report that describes fatal Cases of salmonellosis from blue and gold macaws.

Subcutaneous clostridial infection in Adelie penguins in Hope Bay, Antarctica

During the 2000–2001 breeding season in Hope Bay, Antarctica, two adult Adelie penguins were found dead with lesions compatible with subcutaneous clostridial infection. Clostridiumcadaveris was isolated from the musculature and the subcutaneous tissue of one of these two penguins, whereas Clostridiumsporogenes was isolated from the subcutaneous tissue of the other penguin. Escherichiacoli and Staphylococcus spp. were isolated from both animals. This is the first report of subcutaneous clostridial infection in Antarctic Adelie penguins.