Forgotten giants: Robust climate signal in pollarded trees.

Tree ring records are among the most valuable resources to create high-resolution climate reconstructions. Most climate reconstructions are based on old trees growing in inaccessible mountainous areas with low human activity. Therefore, reconstruction of climate conditions in lowlands is usually based on data from distant mountains. Albeit old trees can be common in humanized areas, they are not used for climate reconstructions. Pollarding was a common traditional management in Europe that enabled trees to maintain great vitality for periods exceeding the longevity of unmanaged trees. We evaluate the potential of pollarded deciduous oaks to record past climate signal. We sampled four pollarded woodlands in Central Spain under continental Mediterranean climate. We hypothesized that pollarded trees have a strong response to water availability during current period without pollarding management, but also in the period under traditional management if pruning was asynchronous among trees. Moreover, we hypothesized that if climate is a regional driver of oak secondary growth, chronologies from different woodlands will be correlated. Pollard oaks age exceeded 500 years with a strong response to Standardized Precipitation-Evapotranspiration Index (SPEI) from 9 to 11 months. Climate signal was exceptionally high in three of the sites (r2 = 0.443-0.655) during low management period (1962-2022). The largest fraction of this climate signal (≈70 %) could be retrieved during the traditional management period (1902-1961) in the three sites where pollarding was asynchronous. Chronologies were significantly correlated since the 19th century for all the studied period, highlighting a shared climate forcing. We identified critical points to optimize pollard tree sampling schema. Our results show the enormous potential of pollarded woodlands to reconstruct hydroclimate conditions in the Mediterranean with a fine spatial grain. Studying pollarded trees is an urgent task, since the temporal window to retrieve the valuable information in pollarded trees is closing as these giants collapse and their wood rots.

You'd better walk alone: Changes in forest composition affect pollination efficiency and pre-dispersal cone damage in Iberian Juniperus thurifera forests.

Changes in land-use patterns are a major driver of global environmental change. Cessation of traditional land-use practices has led to forest expansion and shifts in forest composition. Consequently, former monospecific forests maintained by traditional management are progressing towards mixed forests. However, knowledge is scarce on how the presence of other tree species will affect reproduction of formerly dominant species. We explored this question in the wind-pollinated tree Juniperus thurifera. We hypothesised that the presence of heterospecific trees would have a negative effect on cone production and on the proportion of cones attacked by specialised predators. We assessed the relative importance of forest composition on cone production, seed development and pre-dispersal cone damage on nine paired pure and mixed J. thurifera forests in three regions across the Iberian Peninsula. The effects of forest composition on crop size, cone and seed characteristics, as well as damage by pre-dispersal arthropods were tested using mixed models. Cone production was lower and seed abortion higher in mixed forests, suggesting higher pollination failure. In contrast, cone damage by arthropods was higher in pure forests, supporting the hypothesis that presence of non-host plants reduces damage rates. However, the response of each arthropod to forest composition was species-specific and the relative rates of cone damage varied depending on individual tree crops. Larger crop sizes in pure forests compensated for the higher cone damage rates, leading to a higher net production of sound seeds compared to mixed forests. This study indicates that ongoing changes in forest composition after land abandonment may impact tree reproduction.

Rickettsia conorii is a potent complement activator in vivo and combined inhibition of complement and CD14 is required for attenuation of the cytokine response ex vivo.

Mediterranean spotted fever caused by Rickettsia conorii is a potentially lethal disease characterized by vascular inflammation affecting multiple organs. Studies of R. conorii so far have focused on activation of inflammatory cells and their release of inflammatory cytokines, but complement activation has not been investigated in R. conorii-infected patients. Here, we performed a comprehensive analysis of complement activation markers and the soluble cross-talking co-receptor CD14 (sCD14) in plasma from R. conorii-infected patients. The clinical data were supplemented with ex vivo experiments where the cytokine response was characterized in human whole blood stimulated with R. conorii. Complement activation markers at the level of C3 (C3bc, C3bBbP) and terminal pathway activation (sC5b-9), as well as sCD14, were markedly elevated (p <0.01 for all), and closely correlated (p <0.05 for all), in patients at admission compared with healthy matched controls. All tested markers were significantly reduced to baseline values at time of follow up. Rickettsia conorii incubated in human whole blood was shown to trigger complement activation accompanied by release of the inflammatory cytokines interleukin-1ß (IL-1ß), IL-6, IL-8 and tumour necrosis factor. Whereas inhibition of either C3 or CD14 had only a minor effect on released cytokines, combined inhibition of C3 and CD14 resulted in significant reduction, virtually to baseline levels, of the four cytokines (p <0.05 for all). Our data show that complement is markedly activated upon R. conorii infection and complement activation is, together with CD14, responsible for a major part of the cytokine response induced by R. conorii in human whole blood.

Efficiency of pollination and satiation of predators determine reproductive output in Iberian Juniperus thurifera woodlands.

Fruit production in animal-dispersed plants has a strong influence on fitness because large crops increase the number of seeds dispersed by frugivores. Large crops are costly, and environmental control of plant resources is likely play a role in shaping temporal and spatial variations in seed production, particularly in fluctuating environments such as the Mediterranean. The number of fruits that start to develop and the proportion of viable seeds produced are also linked to the number of flowers formed and the efficiency of pollination in wind-pollinated plants. Finally, large fruit displays also attract seed predators, having a negative effect on seed output. We assessed the relative impact of environmental conditions on fruit production, and their combined effect on seed production, abortion and seed loss through three predispersal predators in Juniperus thurifera L., sampling 14 populations across the Iberian Peninsula. Wetter than average conditions during flowering and early fruit development led to larger crop sizes; this effect was amplified at tree level, with the most productive trees during more favourable years yielding fruits with more viable seeds and less empty and aborted seeds. In addition, large crops satiated the less mobile seed predator. The other two predispersal predators responded to plant traits, the presence of other seed predators and environmental conditions, but did not show a satiation response to the current-year crop. Our large-scale study on a dioecious, wind-pollinated Mediterranean juniper indicates that pollination efficiency and satiation of seed predators, mediated by environmental conditions, are important determinants of reproductive output in this juniper species.

Does plant colour matter? Wax accumulation as an indicator of decline in Juniperus thurifera.

The photosynthesis in evergreen trees living in Mediterranean ecosystems is subjected to multiple climatic stresses due to water shortage and high temperatures during the summer and to low temperatures during the winter. Mediterranean perennials deploy different photoprotective mechanisms to prevent damage to the photosynthetic system. Wax accumulation in leaves is a primary response which by enhancing light scattering in the leaf surface reduces incident radiation in the mesophyll. The existence of high variability in wax accumulation levels between coexisting individuals of a species has a visual effect on colour that provides distinguishable green and glaucous phenotypes. We explored this variability in a Mediterranean evergreen tree Juniperus thurifera (L.) to evaluate the impact of epicuticular wax on optical and ecophysiological properties and on the abundance of photoprotective pigments throughout an annual cycle. Because of light attenuation by waxes, we expected that glaucous phenotypes would lower the need for photoprotective pigments. We evaluated the effect of phenotype and season on reflectance, defoliation levels, photochemical efficiency and photoprotective pigment contents in 20 green and 20 glaucous junipers. Contrary to our expectations, the results showed that glaucous trees suffered from a diminution in photochemical efficiency, but there was no reduction in photoprotective pigments. Differences between glaucous and green phenotypes were greater in winter, which is the most stressful season for this species. Glaucous individuals also showed the highest levels of leaf defoliation. The lower photochemical efficiency of glaucous trees, together with higher defoliation rates and equal or greater number of physiological photoprotective mechanisms, suggests that in spite of wax accumulation, glaucous trees suffer from more severe stress than green ones. This result suggests that changes in colouration in Mediterranean evergreens may be a decline indicator.

Soil seed bank recovery occurs more rapidly than expected in semi-arid Mediterranean gypsum vegetation.

BACKGROUND AND AIMS: Seed banks are critical in arid ecosystems and ensure the persistence of species. Despite the importance of seed banks, knowledge about their formation and the extent to which a seed bank can recover after severe perturbation remains scarce. If undisturbed, soil seed banks reflect a long vegetation history; therefore, we would expect that new soil seed banks and those of undisturbed soils require long periods to become similar with respect to both density and composition. In contrast, if soil seed banks are only a short- to mid-term reservoir in which long-term accumulation constitutes only a tiny fraction, they will recover rapidly from the vegetation. To shed light on this question, we evaluated seed bank formation in a semi-arid gypsum community. METHODS: Soils from 300 plots were replaced with sterilized soil in an undisturbed semi-arid Mediterranean community. Seasonal changes in seed bank density and composition were monitored for 3 years by comparing paired sterilized and control soil samples at each plot. KEY RESULTS: Differences in seed bank density between sterilized and control soil disappeared after 18 months. The composition of sterilized seed banks was correlated with that of the control plots from the first sampling date, and both were highly correlated with vegetation. Nearly 24 % of the seed bank density could be attributed to secondary dispersal. Most seeds died before emergence (66·41-71·33 %), whereas the rest either emerged (14·08-15·48 %) or persisted in the soil (14·59-18·11 %). CONCLUSIONS: Seed banks can recover very rapidly even under the limiting and stressful conditions of semi-arid environments. This recovery is based mainly on the seed rain at small scales together with secondary dispersal from intact seed banks in the vicinity. These results emphasize the relevance of processes occurring on short spatial scales in determining community structure.

Native plant communities in an abandoned Pb-Zn mining area of northern Spain: implications for phytoremediation and germplasm preservation.

Plants growing on metalliferous soils from abandoned mines are unique because of their ability to cope with high metal levels in soil. In this study, we characterized plants and soils from an abandoned Pb-Zn mine in the Basque Country (northern Spain). Soil in this area proved to be deficient in major macronutrients and to contain toxic levels of Cd, Pb, and Zn. Spontaneously growing native plants (belonging to 31 species, 28 genera, and 15 families) were botanically identified. Plant shoots and rhizosphere soil were sampled at several sites in the mine, and analyzed for Pb, Zn and Cd concentration. Zinc showed the highest concentrations in shoots, followed by Pb and Cd. Highest Zn concentrations in shoots were found in the Zn-Cd hyperaccumulator Thlaspi caerulescens (mean = 18,254 mg Zn kg(-1) DW). Different metal tolerance and accumulation patterns were observed among the studied plant species, thus offering a wide germplasm assortment for the suitable selection of phytoremediation technologies. This study highlights the importance of preserving metalliferous environments as they shelter a unique and highly valuable metallicolous biodiversity.

Factors contributing to emergence of Ehrlichia and Anaplasma spp. as human pathogens.

Ehrlichioses and anaplasmoses are caused by alpha-proteobacteria within the family of Anaplasmataceae. These diseases have been known for a long time in veterinary medicine and recently in human medicine. These tick-borne zoonoses are considered as emerging diseases. The first case of human monocytotropic ehrlichiosis occurred in 1986. Human granulocytic anaplasmosis was described as a separate entity in 1994 and ehrlichiosis caused by Ehrlichia ewingii was reported in humans in 1999. The number of cases has been rising steadily due to better diagnostic techniques and better surveillance worldwide. In this review, we will present human and animal ehrlichioses and anaplasmoses as emerging diseases and present candidate(s) for the future.

Carbohydrate storage in five resprouting Florida scrub plants across a fire chronosequence.

Most research analyzing nonstructural carbohydrate (NSC) concentrations on resprouter species in fire-controlled ecosystems has concentrated on how NSC concentrations recover immediately after fire. However, we know little of the effect of long periods without fire on NSC concentrations. In order to assess the effect of different periods of time-since-fire on resprouter species, we studied carbohydrate concentrations (total [NSC], soluble sugars [SS] and nonsoluble sugars [NSS]) in five resprouting species with contrasting trends of abundance across a chronosequence of time-since-fire (0.5-40 yr) in Florida. Carbohydrate concentrations were highest in species with specialized reserve organs. [SS] was mainly explained by factors related to plant size, whereas time-since-fire was the main factor explaining [NSS]. Changes in [NSS] and [NSC] were correlated with the time-since-fire abundance patterns. Variation in [NSS] carbohydrates can be related to the structural development of vegetation, with only those species capable of accessing full light able to accumulate carbohydrates, whereas subordinate plants show reductions in the [NSS] carbohydrate fractions. In areas with long intervals between fires, this carbohydrate reduction could affect subsequent postfire resprouting vigour, although this remains to be confirmed.

Attenuation of Salmonella enterica Serovar Typhimurium by altering biological functions of murein lipoprotein and lipopolysaccharide.

We constructed Salmonella enterica serovar Typhimurium double-knockout mutants in which either the lipoprotein A (lppA) or the lipoprotein B (lppB) gene was deleted from an msbB-negative background strain by marker exchange mutagenesis. These mutants were highly attenuated when tested with in vitro and in vivo models of Salmonella pathogenesis.

Murein lipoprotein is a critical outer membrane component involved in Salmonella enterica serovar typhimurium systemic infection.

Lipopolysaccharide (LPS) and Braun (murein) lipoprotein (Lpp) are major components of the outer membrane of gram-negative enteric bacteria that function as potent stimulators of inflammatory and immune responses. In a previous paper, we provided evidence that two functional copies of the lipoprotein gene (lppA and lppB) located on the chromosome of Salmonella enterica serovar Typhimurium contributed to bacterial virulence. In this study, we characterized lppA and lppB single-knockout (SKO) mutants and compared them with an lpp double-knockout (DKO) mutant using in vitro and in vivo models. Compared to the lpp DKO mutant, which was nonmotile, the motility of the lpp SKO mutants was significantly increased (73 to 77%), although the level of motility did not reach the level of wild-type (WT) S. enterica serovar Typhimurium. Likewise, the cytotoxicity was also significantly increased when T84 human intestinal epithelial cells and RAW264.7 murine macrophages were infected with the lpp SKO mutants compared to the cytotoxicity when cells were infected with the lpp DKO mutant. The level of interleukin-8 (IL-8) in polarized T84 cells infected with the lppB SKO mutant was significantly higher (two- to threefold higher), reaching the level in cells infected with WT S. enterica serovar Typhimurium, than the level in host cells infected with the lppA SKO mutant. The lpp DKO mutant induced minimal levels of IL-8. Similarly, sera from mice infected with the lppB SKO mutant contained 4.5- to 10-fold-higher levels of tumor necrosis factor-alpha and IL-6; the levels of these cytokines were 1.7- to 3.0-fold greater in the lppA SKO mutant-infected mice than in animals challenged with the lpp DKO mutant. The increased cytokine levels observed with the lppB SKO mutant in mice correlated with greater tissue damage in the livers and spleens of these mice than in the organs of animals infected with the lppA SKO and lpp DKO mutants. Moreover, the lppB SKO mutant-infected mice had increased susceptibility to death. Since the lpp DKO mutant retained intact LPS, we constructed an S. enterica serovar Typhimurium triple-knockout (TKO) mutant in which the lppA and lppB genes were deleted from an existing msbB mutant (msbB encodes an enzyme required for the acylation of lipid A). Compared to the lpp DKO and msbB SKO mutants, the lpp-msbB TKO mutant was unable to induce cytotoxicity and to produce cytokines and chemokines in vitro and in vivo. These studies provided the first evidence of the relative contributions of Lpp and lipid A acylation to Salmonella pathogenesis.

Molecular characterization of a glucose-inhibited division gene, gidA, that regulates cytotoxic enterotoxin of Aeromonas hydrophila.

By using a mini-transposon, we obtained two mutated strains of a diarrheal isolate, SSU, of Aeromonas hydrophila that exhibited a 50 to 53% reduction in the hemolytic activity and 83 to 87% less cytotoxic activity associated with the cytotoxic enterotoxin (Act). Act is a potent virulence factor of A. hydrophila and has been shown to contribute significantly to the development of both diarrhea and septicemia in animal models. Subsequent cloning and DNA sequence analysis revealed that transposon insertion occurred at different locations in these two mutants within the same 1,890-bp open reading frame for the glucose-inhibited division gene (gidA). A similar reduction in hemolytic (46%) and cytotoxic (81%) activity of Act was noted in the gidA isogenic mutant of A. hydrophila that was generated by marker exchange mutagenesis. Northern blot analysis revealed that the transcription of the cytotoxic enterotoxin gene (act) was not altered in the gidA transposon and isogenic mutants. However, by generating a chromosomal act::alkaline phosphatase gene (phoA) reporter construct, we demonstrated significantly reduced phosphatase activity in these mutants, indicating the effect of glucose-inhibited division (GidA) protein in modulating act gene expression at the translational level. The biological effects of Act in the gidA mutants were restored by complementation. The virulence of the gidA mutants in mice was dramatically reduced compared to the those of the wild-type (WT) and complemented strains of A. hydrophila. The histopathological examination of lungs, in particular, indicated severe congestion, alveolar hemorrhage, and acute inflammatory infiltrate in the interstitial compartment and the alveolar spaces when mice were infected with the WT and complemented strains. Minimal-to-mild changes were noted in the lungs with the gidA mutants. Taken together, our data indicate for the first time that GidA regulates the most-potent virulence factor of A. hydrophila, Act.

Rickettsial infection in murine models activates an early anti-rickettsial effect mediated by NK cells and associated with production of gamma interferon.

Natural killer (NK) cell activity was significantly increased on days 2-6 of infection in the Rickettsia conorii-infected C3H/HeN mice and on day 2 in the Rickettsia typhi-infected C57BL/6 mice. Depletion of NK cell activity utilizing anti-NK1.1 monoclonal antibody enhanced the susceptibility of normally resistant C57BL/6 mice to infection with R. typhi, and depletion of NK cell activity with antibody to asialo GM1 enhanced the susceptibility of C3H/HeN mice to infection with R. conorii. Serum gamma interferon was increased in R. conorii-infected C3H/HeN mice compared with NK cell-depleted, infected mice during the early course of infection. Additionally, the NK cell activating cytokine IL-12 was elevated in the sera of infected mice during the time period representing enhanced NK cell activity compared with uninfected mice. Thus, it appears that NK cells contribute to the early anti-rickettsial immune response, likely via a mechanism involving gamma interferon.

Critical role of cytotoxic T lymphocytes in immune clearance of rickettsial infection.

Cytotoxic T-lymphocyte (CTL) activity developed against the major infected target cells of rickettsial infections, endothelial cells and macrophages. Spleen cells from mice immune to Rickettsia conorii exerted specific major histocompatibility complex (MHC) class I-matched CTL activity against R. conorii-infected SVEC-10 endothelial cells, with peak activity on day 10. Similarly, spleen cells from Rickettsia australis-immune mice exerted specific CTL activity against an R. australis-infected macrophage-like cell line. Gamma interferon (IFN-gamma) gene knockout mice were more than 100-fold more susceptible to R. australis infection than wild-type C57BL/6 mice. MHC class I gene knockout mice were the most susceptible, more than 50,000-fold more susceptible to a lethal outcome of R. australis infection than wild-type C57BL/6 mice. These results indicate that CTL activity was more critical to recovery from rickettsial infection than were the effects of IFN-gamma. The observation that perforin gene knockout mice were more than 100-fold more susceptible than wild-type C57BL/6 mice indicates that perforin-mediated activity accounts for a large component, but not all, of the CTL-mediated antirickettsial effect. CTL activity was expressed by immune CD8 T lymphocytes. Adoptive transfer of immune CD8 T lymphocytes from IFN-gamma gene knockout mice into R. australis-infected IFN-gamma gene knockout mice dramatically reduced the infectious rickettsial content in the organs, confirming that CD8 T lymphocytes provide immunity against rickettsiae besides that provided by the secretion of IFN-gamma. CTLs appear to be crucial to recovery from rickettsial infection.

Animal model of fatal human monocytotropic ehrlichiosis.

Human monocytotropic ehrlichiosis caused by Ehrlichia chaffeensis is a life-threatening, tick-borne, emerging infectious disease for which no satisfactory animal model has been developed. Strain HF565, an ehrlichial organism closely related to E. chaffeensis isolated from Ixodes ovatus ticks in Japan, causes fatal infection of mice. C57BL/6 mice became ill on day 7 after inoculation and died on day 9. The liver revealed confluent necrosis, ballooning cell injury, apoptosis, poorly formed granulomas, Kupffer cell hyperplasia, erythrophagocytosis, and microvesicular fatty metamorphosis. The other significant histological findings consisted of marked expansion of the marginal zone and infiltration of the red pulp of the spleen by macrophages, interstitial pneumonitis, and increased numbers of immature myeloid cells and areas of necrosis in the bone marrow. Ehrlichiae were detected by immunohistology and electron microscopy in the liver, lungs, and spleen. The main target cells were macrophages, including Kupffer cells, hepatocytes, and endothelial cells. Apoptosis was detected in Kupffer cells, hepatocytes, and macrophages in the lungs and spleen. This tropism for macrophages and the pathological lesions closely resemble those of human monocytotropic ehrlichiosis for which it is a promising model for investigation of immunity and pathogenesis.

Factors affecting establishment of a gypsophyte: the case of Lepidium subulatum (Brassicaceae).

The restriction of vascular plants to gypsum-rich soils under arid or semiarid climates has been reported by many authors in different parts of the world. However, factors controlling the presence of gypsophytes on these soils are far from understood. We investigated the establishment of Lepidium subulatum, a gypsophyte, in a nondisturbed semiarid gypsum-soil landscape in central Spain, both from spatial and temporal perspectives. Over 1400 seedlings were tagged, and their growth and survival were monitored for a 2-yr period. Several biotic and abiotic variables were measured to determine the factors controlling the emergence and early survival. These variables included the cover of annual plants, bryophytes, lichens, litter, gypsum crystals, bare fraction and cover of each perennial plant, and several soil properties (gravel, fine gravel, and fine-earth fraction, conductivity, pH, gypsum content, organic matter and penetrometer soil resistance). Our results support the linkage of gypsophily with some physical properties of the surface crust. Seedlings tended to establish on the gypsum surface crust, and their survival was size dependent, probably as a consequence of the necessity of rooting below the surface crust before summer drought arrives. However, once seedlings emerged, a higher survival rate occurred on the alluvial soils of the piedmont-slope boundary where soil crusts are absent or thinner. We conclude that Lepidium subulatum may be considered a refuge model endemic with a distribution range that occupies a reduced fraction of a wider habitat from which it is probably excluded by competition.

Chemical modification of NADP-isocitrate dehydrogenase from Cephalosporium acremonium evidence of essential histidine and lysine groups at the active site.

NADP-isocitrate dehydrogenase from Cephalosporium acremonium CW-19 has been inactivated by diethyl pyrocarbonate following a first-order process giving a second-order rate constant of 3.0 m-1. s-1 at pH 6.5 and 25 degrees C. The pH-inactivation rate data indicated the participation of a group with a pK value of 6.9. Quantifying the increase in absorbance at 240 nm showed that six histidine residues per subunit were modified during total inactivation, only one of which was essential for catalysis, and substrate protection analysis would seem to indicate its location at the substrate binding site. The enzyme was not inactivated by 5, 5'-dithiobis(2-nitrobenzoate), N-ethylmaleimide or iodoacetate, which would point to the absence of an essential reactive cysteine residue at the active site. Pyridoxal 5'-phosphate reversibly inactivated the enzyme at pH 7.7 and 5 degrees C, with enzyme activity declining to an equilibrium value within 15 min. The remaining activity depended on the modifier concentration up to about 2 mm. The kinetic analysis of inactivation and reactivation rate data is consistent with a reversible two-step inactivation mechanism with formation of a noncovalent enzyme-pyridoxal 5'-phosphate complex prior to Schiff base formation with a probable lysyl residue of the enzyme. The analysis of substrate protection shows the essential residue(s) to be at the active site of the enzyme and probably to be involved in catalysis.

Dual infection with Ehrlichia chaffeensis and a spotted fever group rickettsia: a case report.

Well-documented cases of simultaneous human infection with more than one tick-borne pathogen are rare. To our knowledge only two dual infections have been reported: simultaneous human infection with the agent of human granulocytic ehrlichiosis and Borrelia burgdorferi and simultaneous human infection with B. burgdorferi and Babesia microti (1-2). Rocky Mountain spotted fever has long been known to be endemic in North Carolina; cases of human ehrlichial infection were recognized there soon after Ehrlichia chaffeensis was recognized as an important cause of tick-borne disease in the southeastern United States. Because both Rocky Mountain spotted fever and ehrlichiosis are prevalent in North Carolina, occasional cases of simultaneous human infection by rickettsial and ehrlichial agents would not be surprising; however, no such cases seem to have been reported.

Evaluation of the MycoAKT latex agglutination test for rapid diagnosis of Mycobacterium avium complex infections.

Rapid diagnosis of Mycobacterium avium complex (MAC) bacteremia is important for management of patients with the acquired immunodeficiency syndrome who have disseminated MAC. The purpose of this study was to determine the reliability of the MycoAKT latex agglutination test for direct detection of MAC in positive mycobacterial blood cultures. First, colonies of isolates of previously identified mycobacteria, including 35 MAC, were tested. Of the 55 isolates evaluated, 33 were identified as MAC by the latex test, including 31 of the known MAC and 2 M. chelonae (sensitivity, 88.6%; specificity, 90.0%). Second, broth from 20 ESP II and 20 BACTEC 12B bottles seeded with isolates of MAC were tested. Aliquots from 19 (95%) ESP II cultures and 16 (80%) 12B cultures were positive by the latex test. In phase 3, broth from 115 signal-positive ESP II blood cultures were tested by latex agglutination. Forty-three subcultures from these bottles grew mycobacteria (41 MAC and 2 Mycobacterium tuberculosis complex); the remainder grew no organisms. Broth from 40 of the blood cultures (39 that grew MAC and 1 from which no organisms were recovered) were latex positive; thus, the sensitivity, specificity, and positive and negative predictive values of the latex test for direct identification of MAC in ESP II blood cultures were 95.1, 98.6, 97.5, and 97.3%, respectively. The mean time to detection of MAC was 14.6 days (range, 6-34 days) with the direct latex test, compared with 18.3 days (range, 9-36 days) with subculture and probe (p < 0.05).