Examination of microbial contaminants of emergency showers and eyewash stations

To evaluate the effects of regular flushing, water from fifty emergency eyewash and shower stations was cultured for the presence of potentially pathogenic protozoa, heterotrophic bacteria, and Legionella species. This study also provided the opportunity to evaluate a commercially available molecular assay for the direct detection of Legionella sp in environmental samples. The Perkin Elmer Legionella EnviroAmp polymerase chain reaction (PCR) kit and culture on buffered charcoal yeast extract agar were used to detect Legionella species in water samples. Chemical and physical parameters of station water measured included: pH, hardness, alkalinity, turbidity, conductivity, total chlorine and assimilable organic carbon. Protozoal isolates were identified by classical identification methods, and isolates from the stations were identified as Hartmannella sp, Vexillifera sp, Vahlkampfia sp, Acanthamoeba sp, and Vanella sp. Heterotrophic plate counts ranged from 10² to 10⁶ CFU ml⁻¹ and acridine orange total counts ranged from 10³ to 10⁶ cells ml⁻¹ after regular flushing. PCR and gene probe analysis showed that 89% of the stations (eyewash and shower) were positive for Legionella species by PCR, while 6% of the samples were culture positive. These results indicate that routine flushing alone is not sufficient to control microbial contamination and disinfection must also be included in a routine maintenance program. In addition, regular maintenance, disinfection, and monitoring of emergency eyewash and shower stations is important in preventing potential secondary microbial infections by either direct inoculation or aerosol transmission. Received 02 September 1997/ Accepted in revised form 29 November 1997     

Dermatophytes isolated from laboratory animals

In order to determine the presence of dermatophytes in healthy skin, 200 animals from the animal house of Faculty of Medicine, U.N.A.M., were studied; these were 50 rats, 50 rabbits, 50 mice, and 50 guinea pigs. Out of these 200 animals, 29.5% had positive isolation of Trichophyton mentagrophytes, var. lacticolor. The frequency variation was: rats, 68%; rabbits, 36%; mice, 8%; and guinea pigs, 6%. Male rats and male rabbits, had the higher incidence of positives. The epidemiologic repercussion of these and the significance to use these animals in biomedical investigation is discussed.     

Amoebae from antarctic soil and water.

Samples of soil and water were taken from the McMurdo Sound-Dry Valley region of Antarctica. Of the 70 samples cultured, 22 yielded amoebae capable of clonal growth at 30 degrees C. None of the isolates was pathogenic for mice. Acanthamoeba isolates appeared to show better survival potential than Naegleria isolates.     

A survey of staphylococci isolated from the laboratory gerbil

A microbiological survey of the Mongolian gerbil, Meriones unguiculatus, revealed coagulase-negative staphylococci to be common inhabitants of representative animals derived from three different breeding colonies. The nasal area was most often culture positive, and Staphylococcus xylosus was a predominant species. S. xylosus was the only organism cultured from nasal dermatitis. These organisms were found to be susceptible in vitro to the majority of the antimicrobial agents tested. This survey indicates that the possible role of S. xylosus as an opportunistic pathogen warrants further investigation.     

Heat resistance of Cladosporium isolated from laboratory animal facilities

Heat resistance tests for the saprophyte, Cladosporium, isolated from laboratory animal facilities were carried out. In testing the effects of moderate and high temperature conditions, C. sphaerospermum (C. s) and C. cladosporioides (C. c) were found to grow on media in temperatures less than 32 degrees C, but did not in temperature of 35 degrees C and over. The colony diameter of Cladosporium became smaller as temperature increased. The death time of C. s treated with moist heat was within 12 min at 48 degrees C and that of C. c was within 26 min at 43 degrees C. Both Cladosporium species could not survive for more than 1 min at 55 degrees C. On the other hand, Cladosporium treated with dry heat could not survive more than 69-12 min (C. s) and 39-9.5 min (C. c) at 70-100 degrees C. From these results, it can be seen that Cladosporium was definitely sensitive to heat treatment, and the authors assume that heat is a means of prevention in laboratory animal facilities.     

Giant Amoebae from Colorado Identified by Transplantation Tolerance1

SYNOPSIS. Giant multinucleated amoebae, discovered in Colorado near Ft. Collins and tentatively identified as Pelomyxa carolinensis, were successfully cultured in a manner identical with that used for P. carolinensis. The ultrastructure of the Colorado amoebae, in comparison with that of other large amoebae, was like that of P. carolinensis, but different from those of Amoeba proteus, P. illinoisensis, and P. palustris. Protoplasmic grafts between the Colorado amoebae and P. carolinensis were then exchanged by microsurgery. These grafts were well tolerated, and the recipient amoebae reproduced at a rate comparable to that of non-grafted controls. Other Colorado amoebae received grafts from P. illinoisensis, but all recipients died without cell division a few days after microsurgery. These nutritional, cytologic and transplantation-tolerance data reveal that the amoebae from Colorado are P. carolinensis.     

Serotypes of Pseudomonas aeruginosa isolated from laboratory rats and mice

Pseudomonas aeruginosa isolates cultured from the feces of laboratory rats and mice were serotyped. The fecal samples originated from primary genetic centers, secondary breeding facilities, and research testing facilities operated under contracts from the National Cancer Institute. Eighty-nine percent or 264 of 297 isolates were of serotypes 1, 4, 6, 10, or 11, and of these, 154 (51.8%) isolates were serotypes 6 or 11. In some instances, Pseudomonas aeruginosa serotypes found in animals at a primary genetic center were also found at secondary breeding facilities which had received breeding stock from the primary genetic center. The same serotypes also were found in animals at research-testing laboratories that had received animals from the secondary breeding facilities.     

Monitoring of antibiotic resistance in bacteria isolated from laboratory animals

The drug resistance of microorganisms isolated from laboratory animals never treated with antibiotics is being reported consistently, while the number of laboratory animals used in medicine, pharmacy, veterinary medicine, agriculture, nutrition, and environmental and health science has increased rapidly in Korea. Therefore, this study examined the development of antimicrobial resistance in bacteria isolated from laboratory animals bred in Korea. A total of 443 isolates (7 species) containing 5 Sphingomonas paucimobilis, 206 Escherichia coli, 60 Staphylococcus aureus, 15 Staphylococcus epidermidis, 77 Enterococcus faecalis, 27 Citrobacter freundii, 35 Acinetobacter baumannii were collected from the nose, intestine, bronchus and reproductive organs of ICR mice and SD rats. Of these species, Acinetobacter baumannii and Enterococcus faecalis showed significant antimicrobial resistance according to the minimum inhibition concentration (MIC) in E-test. In case of Acinetobacter baumannii, several isolates showed MIC values 16-128 μg/mL for cefazolin and cefoxitin, and higher resistance (128-512 μg/mL) to nitrofurantoin than that of standard type. Resistance to cefazolin, cefoxitin and nitrofurantoin was detected in 17.14, 20.00, and 8.57% of the Acinetobacter baumannii isolates, respectively. In addition, 44.1% of the Enterococcus faecalis isolates collected from the laboratory animals were resistant to oxacillin concentration of 16-32 μg/mL range, while MIC value of standard type was below oxacillin concentration of 6 μg/mL. These results suggest that in rodent species of laboratory animals, Acinetobacter baumannii are resistance to cefazolin, cefoxitin and nitrofurantoin, whereas those of Enterococcus faecalis were resistance to oxacillin.     

Testate Amoebae from a Cretaceous Forest Floor Microbiocoenosis of France

ABSTRACT. Amber‐preserved shells of testate amoebae often provide as many diagnostic features as the tests of modern taxa. Most of these well‐preserved microfossils are morphologically assignable to modern species indicating either evolutionary stasis or convergent evolution. Here we describe two Lower Cretaceous testate amoebae that are clearly distinguishable from modern species. Centropyxis perforata n. sp. and Leptochlamys galippei n. sp. possessed perforate shells that were previously unknown in these genera. They are preserved in highly fossiliferous amber pieces from the Upper Albian (ca. 100 million years old) of Archingeay/Les Nouillers (Charente‐Maritime, southwestern France). Syninclusions of soil and litter dwelling arthropods and microorganisms indicate a limnetic‐terrestrial microhabitat at the floor of a coastal conifer forest.     

Heat and ozone resistance of Bacillus spores isolated from laboratory animals

Heat resistance of free-spores of 78 Bacillus strains isolated from laboratory animals was examined. Spores of 41 out of 78 strains survived for 320 minutes at 70 degrees C, 27 for 160 min, at 100 degrees C, only one for 20 min. at 110 degrees C by autoclaving, and none for 5 min. at 120 degrees C. D-values at 100 degrees C of 9 strains determined were between 5.03 and 30.06 min. Spores of 9 strains from stock cultures were exposed to ozone gas at various conditions. Ozone resistance of spores was closely dependent upon relative humidity. D-values of the spores tested by treatment with 200 ppm ozone at 60% RH were over 200 min., especially over 1,000 min. in 4 strains, indicating that exposure to ozone at a moderate humidity for 6 hours could not sterilize Bacillus spores. At 90% RH, however, treatment with 200 ppm ozone for 6 hr. might be effective for a routine sterilization in laboratory animal facilities.     

Lipid Composition of Soil Amoebae

SYNOPSIS. Lipid content of axenic cultures of 3 species of soil amoebae was investigated. The strains studied were Acanthamoeba sp., Hartmannella rhysodes and Mayorella palestinensis. No appreciable differences were apparent in the amount of sterols and free fatty acids present in the different strains. The sterols were "fast acting" and were present mostly in free form. There was a difference in the quantities of glycerides, Acanthamoeba sp. containing the highest amount. Sterol of H. rhysodes was isolated and identified as ergosterol by chemical and physical criteria. The implications of these findings are discussed.     

Heat shock protein of HSP70 family revealed in some contemporary freshwater Amoebae and in Acanthamoeba sp. from cysts isolated from permafrost samples

A heat shock protein of HSP70 family was revealed for the first time in trophozoites of Acanthamoeba sp. (strain Am8) excysted from cysts previously isolated from samples of permafrost aging 30,000-35,000 years. The constitutive level of this HSP, shown by immunnoblotting in unstressed trophozoites of the ancient acanthamoebae, much surpassed that in unstressed cells of the three examined species of contemporary freshwater amebae of the genus Amoeba.     

Antibiograms of pathogenic bacteria isolated from laboratory rabbits in Ibadan, Nigeria

Antibiotic sensitivity patterns of 21 bacterial isolates from some clinically ill New Zealand rabbits were evaluated against 12 commonly used antibiotics by the Kirby-Bauer disk-diffusion sensitivity testing method. The 21 isolates consisted of six Bordetella bronchiseptica, eight Pasteurella multocida, four Staphylococcus aureus and three Pseudomonas alcaligenes that were associated with snuffles, pneumonia, otitis media, genital infections and conjunctivitis in these groups of caged rabbits. The four bacteria species were all sensitive to kanamycin, gentamycin and enrofloxacin, with variable sensitivity to the other antibiotics tested. The results of this antibiogram could serve as a field guide in the treatment of very acute bacterial diseases of rabbit prior to the availability of the results of local sensitivity tests. Such sensitivity tests should be reviewed yearly to update this antibiogram.     

Streptobacillus moniliformis isolated from otitis media of conventionally kept laboratory rats.

Streptobacillus moniliformis (Sm) was isolated from the middle ear of two inbred albino rats (strain CAP/Kuv) suffering from murine respiratory mycoplasmosis and purulent bilateral otitis media. The animals were kept under conventional conditions and used for immunological studies. The biochemical pattern of the isolate was identical with that of four other Sm strains of different origin but differed in its ability to lyse erythrocytes in sheep blood agar. This is the first Sm strain with hemolysis described. Pathogenicity of the strain was demonstrated in C57BL/6Han mice known to be susceptible to streptobacillosis. Three of five mice inoculated orally developed characteristic signs of a septic lymphadenitis. In the homologous system and with Sm strain ATCC 49567 as antigen, all five sera showed positive titers in the indirect immunofluorescence assay. Possible improvements in the diagnosis and the role of this "forgotten pathogen" in laboratory animal medicine are discussed.     

Symbiont-Induced Strain-Specific Lethal Effect in Amoebae

The cytosol fraction of a symbiont-bearing strain of Amoeba proteus exerted a lethal effect when injected into symbiont-free amoebae of the original strain. The lethal factor appeared to be a protein with a molecular weight of over 200,000. While the effect of the lethal factor on the nucleus was reversible, the host cytoplasm was permanently damaged so that it could not form a viable cell when combined with a normal nucleus.     

Saprophytic fungi isolated from the hair of domestic and laboratory animals with suspected dermatophytosis

Hair samples from domestic and laboratory animals with suspected dermatophytosis were examined for the presence of saprophytic fungi. A nutritionally poor base medium, developed by the author, was used in the isolation and identification of the saprophytes. Three hundred and ninety-four specimens were examined of which 246 were from dogs, 75 from cats, 30 from horses, 19 from cows, 12 from guinea pigs, 5 from rats, 2 from parakeets, 2 from chinchillas and one each from a goat, a mink and a lesser panda (Ailurus fulgens). Moulds classified in 32 genera were isolated. The commonest in order of frequency were members of the genera Penicillium, Cladosporium, Aspergillus, Mucor, Aureobasidium, Alternaria, Scopulariopsis, Trichoderma and Trichothecium. The yeasts that were isolated were not identified. Aureobasidium pullulans was isolated significantly more often (chi 2 test p less than 0.025) from the dog samples than those from cats, Cladosporium spp. in the samples from dogs than horses, Mucor spp. from the cow samples than horses, Penicillium spp. from the dog samples than those of cats or horses. Skin infections caused by any of the contaminants were not encountered.     

A Cytohesin Homolog in Dictyostelium Amoebae

Background

Dictyostelium, an amoeboid motile cell, harbors several paralogous Sec7 genes that encode members of three distinct subfamilies of the Sec7 superfamily of Guanine nucleotide exchange factors. Among them are proteins of the GBF/BIG family present in all eukaryotes. The third subfamily represented with three members in D. discoideum is the cytohesin family that has been thought to be metazoan specific. Cytohesins are characterized by a Sec7 PH tandem domain and have roles in cell adhesion and migration.

Principal Findings

Dictyostelium SecG exhibits highest homologies to the cytohesins. It harbors at its amino terminus several ankyrin repeats that are followed by the Sec7 PH tandem domain. Mutants lacking SecG show reduced cell-substratum adhesion whereas cell-cell adhesion that is important for development is not affected. Accordingly, multicellular development proceeds normally in the mutant. During chemotaxis secG⁻ cells elongate and migrate in a directed fashion towards cAMP, however speed is moderately reduced.

Significance

The data indicate that SecG is a relevant factor for cell-substrate adhesion and reveal the basic function of a cytohesin in a lower eukaryote.     

Comparative Recoveries of Naegleria fowleri Amoebae from Seeded River Water by Filtration and Centrifugation

Detection of pathogenic Naegleria fowleri in environmental water samples, which is necessary for the prevention of primary amoebic meningoencephalitis, generally requires concentrating the samples. Two concentration techniques, filtration and centrifugation, were used to study the recovery of N. fowleri, in vegetative or cystic form, that had been mixed with the two other thermotolerant Naegleria species, N. lovaniensis and N. australiensis. Counting of amoebae was performed by the most probable number method on 10 water replicates of 100 ml and 10 ml each. With both concentration methods, recovery was better for cysts than for trophozoites (53% ± 21% versus 5% ± 5% by filtration and 57% ± 25% versus 22% ± 5% by centrifugation). The recovery of Naegleria trophozoites by filtration was very low, and centrifugation was significantly better than filtration in recovery of Naegleria trophozoites (22% ± 5% versus 5% ± 5%; P < 0.001). For cysts, however, filtration appeared as efficient as centrifugation, with equivalent values for recovery (53% ± 21% versus 57% ± 25%; P > 0.7). Although the recovery of cysts of N. fowleri obtained by filtration (51% ± 24%) appeared higher than that by centrifugation (36% ± 23%), the difference was not significant (P > 0.1). Both concentration methods have highly variable recovery rates, making accurate quantification of low concentrations (<100/liter) of N. fowleri in the environment difficult.     

Presence of Legionella and Free-Living Amoebae in Composts and Bioaerosols from Composting Facilities

Several species of Legionella cause Legionnaires’ disease (LD). Infection may occur through inhalation of Legionella or amoebal vesicles. The reservoirs of Legionella are water, soil, potting soil and compost. Some species of free-living amoebae (FLA) that are naturally present in water and soil were described as hosts for Legionella. This study aimed to understand whether or not the composting facilities could be sources of community-acquired Legionella infections after development of bioaerosols containing Legionella or FLA. We looked for the presence of Legionella (by co-culture) and FLA (by culture) in composts and bioaerosols collected at four composting facilities located in southern Switzerland. We investigated the association between the presence of Legionella and compost and air parameters and presence of FLA. Legionella spp. (including L. pneumophila) were detected in 69.3% (61/88) of the composts and FLA (mainly Acanthamoeba, Vermamoeba, Naegleria and Stenamoeba) in 92.0% (81/88). L. pneumophila and L. bozemanii were most frequently isolated. FLA as potential host for Legionella spp. were isolated from 40.9% (36/88) of the composts in all facilities. In Legionella-positive samples the temperature of compost was significantly lower (P = 0.012) than in Legionella-negative samples. Of 47 bioaerosol samples, 19.1% (9/47) were positive for FLA and 10.6% (5/47) for L. pneumophila. Composts (62.8%) were positive for Legionella and FLA contemporaneously, but both microorganisms were never detected simultaneously in bioaerosols. Compost can release bioaerosol containing FLA or Legionella and could represent a source of infection of community-acquired Legionella infections for workers and nearby residents.