Assessment of the effect of Enteromorpha prolifera on bacterial community structures in aquaculture environment.

In recent years, Enteromorpha prolifera blooms had serious impacts on costal environments and fisheries in China. Nevertheless, the effects of E. prolifera on microbial ecology remain unknown. In this study, for the first time, an Illumina sequencing analysis was used to investigate bacterial communities in source water, aquaculture ponds with E. prolifera, and an aquaculture pond in which E. prolifera -free. Principal coordinate and phylogenic analyses revealed obvious differences among the bacterial communities in the pond water with and without E. prolifera. Abundant bacterial taxa in the E. prolifera-containing pond were generally absent from the pond without E. prolifera. Interestingly, pond water with E. prolifera was dominated by Actinomycetales (> 50%), as well as by anaerobic bacteria in the underlying sediment (Desulfobacterales and Desulfuromonadales (> 20%). Pond water in which E. prolifera-free was dominated by Rhodobacterales (58.19%), as well as aerobic and facultative anaerobic bacteria in the sediment. In addition, the ecological functions of other dominant bacteria, such as Candidatus Aquiluna, Microcella spp., and Marivita spp., should be studied in depth. Overall, massive growth of E. prolifera will have serious effects on bacterial communities, and, thus, it will have an important impact on the environment. The novel findings in this study will be valuable for understanding green tides.

Assessment of bacterial and fungal growth on natural substrates: consequences for preserving caves with prehistoric paintings.

The most representative bacterium (Pseudonocardia sp.) and fungus (Fusarium sp.) from the microbial communities of a cave containing paleolithic paintings were isolated and their growth on natural substrates assessed. Growth was tested at the in situ and optimal, laboratory growth temperature. Development was analyzed with and without supplemented nutrients (glucose, ammonium, phosphate, peptone). Results showed that the assayed bacterium on natural substrate was able to develop best at in situ temperature and the addition of organic nutrients and/or phosphate enhanced its growth. The growth of the assayed fungus, however, was limited by low temperature and the availability of ammonium. These results confirm a differential behavior of microorganisms between the laboratory and the natural environments and could explain previous invasion of fungi reported for some caves with prehistoric paintings.

Utility of helical computed tomography to evaluate the invasion of actinomycetoma; a report of 21 cases.

BACKGROUND: Actinomycetoma is a chronic infection caused by several aerobic actinomycetes; it is a relatively frequent condition in tropical countries like Mexico. It is important to be aware of the extension and depth of the disease (bone and visceral) to make the prognosis and select treatment. OBJECTIVES: Our objective was to evaluate actinomycetomas using helical computed tomography (HCT) as well as its three-dimensional (3D) reconstruction. MATERIAL AND METHODS: Prospective study of clinically and microbiologically proven cases of actinomycetomas, all of them recently diagnosed and untreated or unresponsive to various treatments. All patients underwent simple and contrast HCT with various helical slices of the involved zones. Then three-dimensional reconstructions on the sagittal and coronal planes were made. RESULTS: Twenty-one patients with actinomycetomas were included, 19 males and two females, with a mean age of 35.5 years and mean duration of disease of 4.1 years. The disease was located in the lower limbs in 81%, and in the upper limbs and trunk in 19%. Twenty of the 21 cases were caused by Nocardia brasiliensis and one by Actinomadura madurae. In all patients the disease was localized to the skin and subcutaneous tissue; 76.2% had muscular involvement; 23.8% visceral involvement; 9.5% had bone involvement and 9.5% vascular involvement. The affected area was determined in each case. CONCLUSIONS: HCT provides precise information about the grade of invasion at diverse levels such as visceral, muscular and vascular systems, and the calculation of the affected area.

Evaluation of several modifications of an ecometric technique for assessment of media performance.

Recovery of Listeria monocytogenes 101M, Jonesia denitrificans, salmonellae, and Pediococcus sp. NRRL B-2354 across nine media was evaluated with three modified versions of an ecometric method. Two approaches involved the use of broth cultures (10(8) to 10(9) CFU/ml) of individual strains and either large (10-microl) or small (1-microl) presterilized plastic loops. The third approach involved precultured slants and the inoculation of media with presterilized plastic inoculating needles (10(4) CFU per needle). Absolute growth indices (AGIs) were compared. No significant differences (P < 0.05) between methods were found when tryptic soy agar supplemented with 0.6% yeast extract (TSAYE) was used for the recovery of L. monocytogenes, J. denitrificans, Pediococcus sp. NRRL B-2354, and Salmonella spp. However, the small loop-broth technique recovered significantly fewer Salmonella enterica Typhimurium DT104 and Salmonella Senftenberg 775W cells than the other two techniques did. The performance of each individual bacterial strain on each of nine media was assayed. The recovery of L. monocytogenes was excellent (AGI > 4.8) with TSAYE, PALCAM, modified Oxford medium (MOX), and Baird-Parker agar and slight with modified PRAB (AGI = 0.4) and deMan Rogosa Sharpe (MRS) agar (< 0.1), and the organism was not recovered with the remaining media (modified lysine iron agar [MLIA], xylose lysine desoxycholate [XLD] agar, and xylose lysine tergitol 4 [XLT4] agar). The recovery of J. denitrificans with TSAYE and MOX was excellent, significantly better than that achieved with PALCAM (AGI = 3.0), but the organism was not recovered with Baird-Parker agar or with the other media tested. The recovery of Pediococcus sp. NRRL B-2354 was excellent with TSAYE and modified PRAB medium > Baird-Parker agar > acidified MRS agar, but the organism was not recovered with any of the other media tested. The best recovery of S. enterica Typhimurium DT104 was achieved with TSAYE > MLIA > or = XLD agar > or = XLT4 agar > Baird-Parker > PALCAM, MOX, acidified MRS agar, modified PRAB, and MRS agar. The best recovery of Salmonella Senftenberg 775W was achieved with TSAYE, MLIA, and XLD agar > XLT4 agar, but the organism was not recovered with the other media evaluated.

Assessment of chitin decomposer diversity within an upland grassland.

The breakdown of chitin within an acidic upland grassland was studied. The aim was to provide a molecular characterisation of microorganisms involved in chitin degradation in the soil using soil microcosms and buried litter bags containing chitin. The investigation involved an examination of the effects of liming on the microbial communities within the soil and their chitinolytic activity. Microcosm experiments were designed to study the influence of lime and chitin enrichment on the grassland soil bacterial community ex situ under controlled environmental conditions. Bacterial and actinomycete counts were determined and total community DNA was extracted from the microcosms and from chitin bags buried at the experimental site. PCR based on specific 16S rRNA target sequences provided products for DGGE analysis to determine the structure of bacterial and actinomycete communities. Chitinase activity was assessed spectrophotometrically using chitin labelled with remazol brilliant violet. Both liming and chitin amendment increased bacterial and actinomycete viable counts and the chitinase activity. DGGE band patterns confirmed changes in bacterial populations under the influence of both treatments. PCR products amplified from DNA isolated from chitin bags were cloned and sequenced. Only a few matched known species but a prominent coloniser of chitin proved to be Stenotrophomonas maltophilia.

Economic aspects of outbreaks of dermatophilosis in first-calving cows in nine herds of dairy cattle in Israel.

Outbreaks of dermatophilosis in nine herds of dairy cattle in Israel were studied retrospectively. The outbreaks might have been associated with the exposure of the animals to intense showering during the spring and summer months of May to October, in addition to calving and postcalving stress. The acute exudative form of dermatophilosis occurred in the first-calving cows during the six weeks after they calved. The morbidity rate ranged from 10.0 to 66.6 per cent and the period of morbidity ranged from eight to 19 days. Mortality or culling occurred in four of the herds and ranged from 1.6 to 17.1 per cent. The disease also resulted in a gradual loss of condition, a decrease in milk production and marked increases in the somatic cell counts in milk.

Clinical application of PCR-restriction enzyme pattern analysis for rapid identification of aerobic actinomycete isolates.

The accuracy and practicality of PCR-restriction enzyme pattern analysis (PRA) for routine identification of aerobic actinomycete clinical isolates were evaluated for 299 cultures submitted to the Mycobacteria/Nocardia Laboratory at the University of Texas Health Center at Tyler. PRA identification using an amplified 439-bp segment (amplicon) of the 65-kDa heat shock protein gene was compared to identification by traditional methods, including growth characteristics, susceptibility patterns, biochemical testing, and high-performance liquid chromatography analysis. Microbiological examination of six cultures ruled out aerobic actinomycetes, and they were omitted from the study. Amplicons were analyzed with BstEII, HaeIII, MspI, HinfI, and BsaHI. When necessary, AciI, HhaI, and NarI were also used. From March 1995 through May 1997 (27 months), 274 of the remaining 293 (93.5%) isolates were accurately identified by PRA. Major diagnostic groups included 170 mycobacteria, 93 nocardiae, and 30 other aerobic actinomycetes. Mixed cultures were readily recognized by PRA, including a wound culture that contained two Nocardia taxa that were indistinguishable morphologically. Mycobacterium mucogenicum was identified in three cultures heavily contaminated with gram-positive cocci. The 19 isolates that produced PRA patterns that did not match those in the current PRA database were differentiated into 8 Mycobacterium species and 11 other aerobic actinomycetes by the presence or absence of BstEII recognition sites. Identification of 15 of these 19 isolates was also equivocal by traditional methods. PRA results were reportable within 2 to 5 working days and were as accurate as and faster and less expensive to obtain than those of traditional methods.

The medically important aerobic actinomycetes: epidemiology and microbiology.

The aerobic actinomycetes are soil-inhabiting microorganisms that occur worldwide. In 1888, Nocard first recognized the pathogenic potential of this group of microorganisms. Since then, several aerobic actinomycetes have been a major source of interest for the commercial drug industry and have proved to be extremely useful microorganisms for producing novel antimicrobial agents. They have also been well known as potential veterinary pathogens affecting many different animal species. The medically important aerobic actinomycetes may cause significant morbidity and mortality, in particular in highly susceptible severely immunocompromised patients, including transplant recipients and patients infected with human immunodeficiency virus. However, the diagnosis of these infections may be difficult, and effective antimicrobial therapy may be complicated by antimicrobial resistance. The taxonomy of these microorganisms has been problematic. In recent revisions of their classification, new pathogenic species have been recognized. The development of additional and more reliable diagnostic tests and of a standardized method for antimicrobial susceptibility testing and the application of molecular techniques for the diagnosis and subtyping of these microorganisms are needed to better diagnose and treat infected patients and to identify effective control measures for these unusual pathogens. We review the epidemiology and microbiology of the major medically important aerobic actinomycetes.