Host range differentiation of spore-positive and spore-negative strain types of Frankia in stands of Alnus glutinosa and Alnus incana in Finland

Host compatibility of different spore-positive (Sp⁺)and spore-negative (Sp^?) strain types of Frankia from alder stands in Finland was studied in Modulation tests with hydrocultures of Alnus glutinosa (L.) Gaertner, A. incana (L.) Moench and A. nitida Endl. Root nodules and soil samples from stands of A. incana (Lammi forest and Hämeenlinna forest) were dominated by Sp ⁺ types of Frankia (coded AiSp⁺ and AiSp⁺ H. respectively), which caused effective root nodules in test plants of A. incana, but failed to induce nodules in A. nitida. In A. glutinosa Frankia strain types AiSp ⁺ and AiSp ⁺ H caused small, ineffective root nodules with sporangia (coded Ineff ^?), which were recognized by the absence or near absence of vesicles in the nodule tissue. Ineffective nodules without sporangia (coded Ineff ^?) were induced on A. glutinosa with soil samples collected at Lammi swamp. The spore-negative strain type of Frankia was common in root nodules of A. glutinosa in Finland (Lammi swamp) and caused effective Sp^? type root nodules (coded AgSp ^?) in hydrocultures of A. incana, A. glutinosa and A. nitida. A different Sp ⁺ strain type of Frankia. coded AgSp⁺ Finland, was occasionally found in stands of A. glutinosa. It was clearly distinguished from strain type AiSp ⁺ by the ability to produce effective nodules on both A. glutinosa and A. incana. The nodulation capacities of soil and nodule samples were calculated from the nodulation response in hydrocutlure and served as a measure for the population density of infective Frankia particles. Sp ⁺ nodules from both strain types had equal and high nodulation capacities with compatible host species. The nodulation capacities of Sp type root nodules from A. glutinosa were consistently low. High frequencies of Frankia AiSp⁺ and AiSp⁺ H were found in the soil environment of dominant AiSp ⁺ nodule populations on A. incana. The numbers of infective particles of this strain type were insignificant in the soil environment of nearby Sp ^? nodule populations on A. glutinosa and in the former field at Hämeen-linna near the Sp⁺ nodule area in Hämeenlinna forest. Strain type AgSp^? had low undulation capacity in the soil environment of both A. incana and A. glutinosa stands, Explanations for the strong associations between Frankia strain types AiSp⁺ and AiSp ^? H and A. incana and between strain type AgSp^? and A. glutinosa are discussed in the light of host specificity and of some characteristics of population dynamics of both strain types. The possible need to adapt the concept of Frankia strain types Sp ⁺ and Sp ^? to strains with some variation in spore development was stressed by the low potentials of strain type AiSp ⁺ H to develop spores in symbioses with hydrocultures of A. incnna.     

Characterization and isolation of melanocyte progenitors from mouse embryos

Whole mount immunohistochemistry and flow cytometry have been used to determine the morphological and molecular features that distinguish melanoblasts from surrounding cells. Whole mount immunohistochemistry of mouse embryos using anti-c-Kit monoclonal antibody revealed two distinct types of c-Kit⁺ cells; one dendritic and the other round in shape. The distribution of c-Kit⁺ dendritic cells in 12.5 days postcoitem embryos correlated well with that of tyrosinase-related protein-2 expression, while the distribution of c-Kit⁺ round cells overlaps that of CD45⁺ cells. This observation suggests that melanoblasts are distinguishable from other c-Kit⁺ cells by their dendritic shape. Mice homozygous for the steel-Dickie mutation (Sl^d/Sl^d) were analyzed to further distinguish melanoblasts from hematopoietic progenitor cells. Sl^d/Sl^d mice are unpigmented but contain hematopoietic cells, although reduced in number. Although no c-Kit⁺ dendritic cells were detectable in the Sl^d/Sl^d embryos, a significant number of c-Kit⁺ round cells were present in the same embryos. To further analyze characteristic features of melanoblasts, c-Kit⁺CD45^? and c-Kit⁺CD45⁺ cells were isolated from dissociated embryonic skin by fluorescent activated cell sorter and the expression of TRP2 melanogenic enzyme was analyzed. Consistent with histological analysis, most c-Kit⁺CD45^? cells were TRP2⁺.c-Kit⁺CD45⁺ cells failed to express TRP2. These results show that most of the melanoblasts are c-Kit⁺TRP2⁺CD45^? dendritic cells and can be discriminated from other cells by flow cytometry or by their morphology.     

Distribution of blood groups among indians in South America. VI. In Paraguay

This paper on the distribution of hereditary factors in the blood of Indians in South America, reports the results of tests made on samples procured from Paraguayan Indians. Specimens from putatively full-blood persons were obtained from the following tribes: 88 Chamacoco, 36 Moro, 85 Chulupi, 207 Lengua, 100 Toba, 20 Yam Lengua, and 51 Guayaki, These 587 Samples were tested for factors in the A-B-O, M-N-S-s, P. Rh-Hr, Lutheran, Kell-Cellano, Lewis, Duffy, Kidd, and Diego systems. Serum samples were tested for haptoglobins and transferrins. He molysates, prepared from whole blood, were tested for hemoglobin types. The results are presented on appropriate tables as number and per cent of phenotypes for the various blood group antigens and their calculated allele frequencies. Locations of the populations from which blood samples were procured are listed on the tables and shown on a map (fig. 1). Of the 587 samples all except two Chamacoco belonged to group O. High frequencies are reported generally for M, s, P, R¹ (CDe), R² (cDE), k (100%) and Fy alleles in Paraguayn Indians. Low frequencies were generally reported for N, S, r (cde) and R° (cDe) alleles. There was a wide variation in frequencies for Di, Jk, and haptoglobin Hp¹. All tested for transferrins were classified as Tf C and all contained hemoglobin (A) as a major component. The following antigens were completely absent: Mi^a, Vw, p, P^k, r^y (CdE), K, and Le¹. Most notable is the unusual distribution of hereditary blood antigens among the Guayaki and Moro. The Guayaki had 100% P₁ and Fy^a; they were higher in R° (cDe), R¹ (CDe), and Jk^a; and lower in R² (cDE) and Hp¹ genes than other Indians; and Di was absent. The Guayaki differed from the other Indians also in having fair skin. The Moro were lower in the P¹ and Jk gene frequencies than is usually found in Amerinds, and the Di gene was absent. The Chamacoco also had an exceptionally low frequency for the P¹ gene (0.261).     

Erythromycin resistant mutants of Bacillus subtilis

Summary Erythromycin resistant (ery ^r) mutants were isolated from Bacillus subtilis ATCC 6633. The composition of ribosomal proteins were analyzed for thirteen such ery ^r-mutants with chromatography on a carboxymethyl cellulose (CMC) column. The 50s subunit from all of the ery ^r-mutants was found to contain the altered 50d protein. The ribosomes prepared from the ery ^r-mutants did not show in vitro alteration of the ability to combine with erythromycin.     

Distribution of hereditary factors in the blood of Indians of the Gila River, Arizona

This paper reports the distribution of blood groups, A-B-H secretors, haptoglobins, transferrins and hemoglobin types among Indians of the Gila River Valley in Arizona. Specimens were procured from the following putative full-bloods: 909 Pima, 37 Papago, and 124 Maricopa; and from the following known mixed-bloods: Pima-Papago 134, Pima-Maricopa 26, Pima-Other Indian 41, Pima-Caucasian 33. These 1304 samples were tested for factors in the A-B-O, M-N-S-s, P, Rh-Hr, Lutheran, Kell-Cellano, Lewis, Duffy, Kidd and Diego blood group systems, and for additional blood factors (Wr^a), Do^a, Vel, Yt^a, Co^a, Gy^a, Sav, and L. W. Serum samples were tested for haptoglobins and transferrins. Hemolysates, prepared from whole blood, were tested for hemoglobin types. The results are presented on appropriate tables as number and per cent of phenotypes for the various blood group antigens and their calculated allele frequencies. Locations of the populations from which blood samples were procured are shown on a map (fig. 1). Tests made by earlier workers on the blood of Arizona Indians and related tribes are presented for comparison and discussed. The usual high frequencies for allele O reported in Amerinds was found among the putatively full-blood Gila Indians; the 124 Maricopa presented the maximum frequency of 1.000. High frequencies were reported generally for M, s, P¹, R¹ (CDe), R² (cDE), k (100%) Fy, and Do^a alleles. Low frequencies were reported for N, S, r (cde), R° (cDe), fy, Le¹w and Di^a (Pima only). There was a wide variation in frequencies for jk, and Hp¹, and there were 17 Transferrin Tf B₁C observed in 270 Pima samples tested. All the remaining were classified as Tf C except two Tf B;C from mixed-bloods. All samples tested for Vel, Yt^a, Co^a, Sav, and Hemoglobin (A) showed the maximum frequency (1.000) for their genes. The following antigens were completely absent: Lu^a, Mi^a, Vw, Mt^a, p, P^k, r^y (CdE), K, and Wr^a. The results of this study suggests that the Papago tribe presents fewer genes of non-Indian origin than the Pima, and the Maricopa least of the three populations.     

Asymmetric crossing over in the spontaneous formation of large deletions in the tonB-trp region of the Escherichia coli K-12 chromosome.

The chromosomal tonB gene of Escherichia coli was used as a target for the detection of spontaneous deletion mutations. The deletions were isolated in both recA ⁺ and recA ⁻ cells, and mutants carrying large deletions were identified because they also lacked part or all of the trp operon. The frequencies of tonB-trp deletion were 1.79 × 10⁻⁹ and 1.09 × 10⁻⁹ for recA ⁺ and recA ⁻ cells, respectively. We analyzed 12 deletions from recA ⁺ and 10 from recA ⁻ cells by cloning and direct sequencing. The deletions ranged in size from 5612 bp to 15142 bp for recA ⁺ and from 5428 bp to 13289 for recA ⁻ cells. Three deletions from recA ⁺ cells and five deletions from recA ⁻ cells were found to have occurred between short sequence repeats at the termini of the deletion, leaving one copy of the repeat in the mutant sequence. Seven deletions from recA ⁺ cells and three deletions from recA ⁻ cells did not have repeats at their termini; in these cases, the DNA sequences that are adjacent to the deletion termini in the wild-type are characterized by short (2–4 bp) repeats. From these results, a model is presented for the generation of deletion mutations which involves formation of an asymmetric crossover mediated by repeated sequences of 2- to 4-bp. Received: 14 September 1998 / Accepted: 22 December 1998     

Detection of cholera (ctx) and zonula occludens (zot) toxin genes in Vibrio cholerae O1, O139 and non-O1 strains

Vibrio cholerae O1 and V. cholerae non-O1 strains isolated from environmental samples collected in São Paulo, Brazil, during cholera epidemics and pre-epidemic periods were examined for the presence of toxin genes. V. cholerae O1 strains isolated from clinical samples in Peru and Mexico, and V. cholerae O139 strains from India were also examined for the presence of ctx (cholera toxin gene) and zot (zonula occludens toxin gene) by polymerase chain reaction (PCR). A modified DNA-extraction method applied in this study yielded satisfactory recovery of genomic DNA from vibrios. Results showed that strains of V. cholerae O1 isolated during the preepidemic period were ctx ^-/zot ^- whereas strains isolated during the epidemic were ctx ⁺/zot ⁺. All V. cholerae non-O1 strains tested in the study were ctx ^-/zot ^-, whereas all V. cholerae O139 strains were ctx ⁺/zot ⁺. Rapid detection of the virulence genes (ctx and zot) can be achieved by PCR and this can serve as an important tool in the epidemiology and surveillance of V. cholerae.     

Salt-tolerance in plants. I. Ions, compatible organic solutes and the stability of plant ribosomes

Abstract Polysomes and ribosomes recovered from a number of plant species were tested for stability when incubated at 25°C in salt solutions in the absence of ATP and initiation factors. Stability was assessed by sucrose density gradient analysis. The stability was inversely proportional to salt concentrations above 125 mol m⁻³ KCl. Polysomes were less stable in the presence of Na⁺ than K⁺ salts, and were much less stable in Cl⁻ than in acetate salts. Polysomes from Triticum aestivum. Hordeum vulgare, Capsicum annuum, Helianthus annuus. Pisum sativum, Atriplex nummularia, Beta vulgaris, Cladophora sp., Enteromorpha sp. and Corallina cuvieri were similarly sensitive to KCl. Polysomes from Ulva lactuca were more sensitive than the other species. Cytoplasmic and plastid polysomes from T. aestivum were similarly unstable in 500 mol m⁻³ KCl. Unprogrammed ribosomal subunit couples from T. aestivum, B. vulgaris and U. lactuca showed Mg²⁺-dependent conformational instability and dissociation in KCl. Slight differences in ribosomal stability were observed between species, but these were unrelated to the salt tolerances of the plants. The ‘compatible’ organic solutes, glycinebetaine and proline, failed to reduce ion-induced instability. Ribosome yield and polysome profiles were similar in leaves of B. vulgaris containing significantly different levels of both Na⁺ and Cl⁻ after growth in media containing 50 or 200 mol m⁻³ NaCl. The results are consistent with the hypothesis that plants maintain a cytoplasmic solute environment that is compatible with ribosomal stability.     

Identity and virulence properties of Aeromonas isolates from diseased fish, healthy controls and water environment in China

Aims: To investigate the species distribution in Aeromonas isolates from diseased fish, healthy controls and water environment in China; to evaluate the frequency of the aerolysin (aer), cytotonic enterotoxin (alt), cytotoxic enterotoxin (act), temperature‐sensitive protease (eprCAI) and serine protease (ahp) genes in Aeromonas isolates; and to determine the potential pathogenicity of these isolates. Methods and Results: Two hundred and two Aeromonas isolates from diseased fish (n = 42), healthy fish (n = 120) and water environment (n = 40) in China were identified to species levels based on sequencing of the housekeeping gene gyrB, while the distribution of five virulence factors, including aer, alt, act, eprCAI and ahp, was investigated by PCR. Aeromonas veronii (25/42; 60%) and Aeromonas hydrophila (14/42; 33%) were the species most commonly isolated from diseased fish, while Aer. veronii was the most common species in healthy fish (90/120; 75%) and water samples (25/40; 62·5%). All the five virulence genes were present in 9% (19/202), among which 10 strains were from diseased fish and nine were identified as Aer. hydrophila. For the strains carrying five virulence genes, the average 50% lethal doses (LD_50s) of strains from diseased fish were lower when compared with the strains from healthy fish and water environment. Conclusions: Aeromonas veronii is the most common species, but no significant difference exists in the isolates obtained from diseased fish and from healthy fish. However, Aer. hydrophila isolates were significantly more frequent from diseased fish than from healthy fish. aer⁺alt⁺act⁺eprCAI⁺ahp⁺ was more frequent virulence genotype in Aeromonas isolates from diseased fish than from healthy fish and water environment, and the aer⁺alt⁺act⁺eprCAI⁺ahp⁺ isolates were more virulent to zebrafish comparing to the other genetic profiles. Significant and Impact of the Study: Aeromonas species in aquatic environments are various and have considerable virulence potential, and therefore, there is a need for more careful and intensive epidemiology studies.     

Distribution of hereditary blood groups among Indians in South America. I. In Ecuador

Blood specimens were procured from 658 Quechua, 36 Colorado, 233 Jivaro, 244 Cayapa, and 48 Secoya Indians of Ecuador. These were examined for antigens in the A-B-O, M-N-S-s, P, Rh-Hr, Lutheran, K-k, Lewis, Duffy and Kidd systems and for Diego (Di^a), Wright (Wr^a), and Berrens (Be^a) agglutinogens as well. Hemolystes were prepared and studied for hemoglobin types and the serum samples were tested for haptoglobins and transfserrins. Gene frequencies are high for O, M, s, R¹, (CDe), R² (cDE), Lu^b, k, Kp^b, Le^b and Fy^a; and low or absent for A, B, N, S, Mi^a, Vw, Mt^a, R⁰ (cDe), V (ce^s), Lu^a, K, Kp^a, Le^a, Fy^b, Js^a, Wr^a and Be^a. The Diego (Di^a) gene is present but its frequency varies greatly from tribe to tribe. Gene frequency Hp¹ is well within the range previously reported for Indians in Middle America excepting the Colorado in which population the frequency of 0.889 is unusually high. All 723 serum specimens tested for transferrins were C or CD. No D or BC types were found. All Ecuadorian Indian bloods tested electrophoretically contained only hemoglobin (A) as a major component.     

Distribution of hereditary blood groups among Indians in South America. II. In Peru

Blood specimens were procured from the following putatively pure Indians of the Peruvian rain forest: 90 Piro and 89 Campa on the Urubamba and Tambo rivers, 142 Shipibo and 14 Isconahua on the Rio Ucayali near Yarinacocha, 151 Aguaruna at Santa Maria de Nieva, where the Marañon and Nieva rivers join, and from 122 Ticuna and 9 Yagua near the Brazilian border on the Amazon. Specimens from highland Indians were obtained from 93 Aymará and 181 Quechua at Puno and environs. These 891 specimens were tested for antigens in the A-B-O, M-N-S-s, P, Rh-Hr, Lutheran, K-k, Lewis, Duffy, Kidd, and Diego (Di^a) systems, and for the Wright (Wr^a) aglutinogen. Serum samples from these bloods were tested for haptoglobins and transferrins and hemolysates were prepared and examined for hemoglobin types. Results for these tests with claculated gene frequencies are presented, for the most part, on appropriate tables. A map is included to show the locations of the populations from which blood samples were procured. As in South American Indians generally, frequencies are high for the O gene it being the only gene of the ABO system which appears in isolated jungle populations and the Aymará. Gene frequencies are usually high also for M, s, R¹ (CDe), R² (cDE), Lu^b, k, Le^H, and Fy^a; and low or absent for A, B, N, S, Mi^a, Vw, R^o (cDe), r (cde), Lu^a, K, Le¹, Fy^b, and Wr^a. The Diego (Di^a) gene is present but varies greatly in frequencies among tribes. Hp¹ gene frequencies vary from 0.44 to 0.69 among the Peruvian Indians tested. Transferrin CD was encountered in only one population i.e., in 3 of 86 Piro (gene frequency Tf^D= 0.02). All others were C. All Peruvian Indian bloods tested electrophoretically contained only hemoglobin (A) as a major component.     

A comparison of isoprene and monoterpene emission rates from the perennial bioenergy crops short‐rotation coppice willow and Miscanthus and the annual arable crops wheat and oilseed rape

Biogenic volatile organic compounds (BVOC) emissions from bioenergy crops may differ from those of conventional crops. We compared emission rates of isoprene and a number of monoterpenes from the lignocellulosic bioenergy crops short‐rotation coppice (SRC) willow and Miscanthus, with the conventional crops wheat and oilseed rape. BVOC emission rates were measured via dynamic vegetation enclosure and GC‐MS analysis approximately monthly between April 2010 and August 2012 at a location in England and from SRC willow at two locations in Scotland. The largest BVOC emission rates were measured from willow in England and varied between years. Isoprene emission rates varied between μg g^?1 h^?1. Of the monoterpenes detected from willow, α‐pinene emission rates were highest (μg g^?1 h^?1), followed by μg g^?1 h^?1 for δ‐3‐carene, μg g^?1 h^?1 for β‐pinene and μg g^?1 h^?1 for limonene. BVOC emission rates measured in Scotland were much lower. Low emission rates of isoprene and α‐pinene were measured from Miscanthus in 2010 (μg g^?1 h^?1 and μg g^?1 h^?1, respectively) but were not detected in subsequent years. Emission rates from wheat of isoprene were negligible but relatively high for monoterpenes (μg g^?1 h^?1 and μg g^?1 h^?1 for α‐pinene and limonene, respectively). No significant emission rates of BVOCs were measured from oilseed rape. The measured emission rates followed a clear seasonal trend. Crude extrapolations based solely on data gathered here indicate that isoprene emissions from willow could correspond to 0.004–0.03% (UK) and 0.76–5.5% (Europe) of current global isoprene if 50% of all land potentially available for bioenergy crops is planted with willow.     

Study of Microflora in Malaysian Dried Fishes and Their Decontamination by Gamma-irradiation

The distribution of microorganisms in 10 samples of salted dried fish and the effects of irradiation of them were studied. The total aerobic bacteria in commercial dried fish were determined to be from 2 × 10⁴ to 3 × 10⁶ per gram. Mold counts were 1 × 10² to 7 × 10³ per gram with a lower amount of yeasts. In spoiled dried fish, total aerobic bacteria were determined to be 4× 10⁶ or 1 × 10⁷ per gram with a few yeasts. Coliforms were not isolated on MacConkey agar plates from any of the samples. The predominant bacteria occurring in spoiled dried fish were Pediococcus halophilus, Vibrio costicola and Planococcus sp. More than 50% of the molds consisted of the Aspergillus niger group, whereas lower amounts of the A. flavus, A. fumigatus and A. ochraceus groups, Penicillium chrysogenum series, etc. were also isolated from many samples of dried fish. All kinds of putrefactive microorganisms were radiation sensitive, and a dose of ca. 500 krad appears to be sufficient for extension of the shelf-life of dried fish from 2 to 4 times.     

Endocrinological investigation into the reproductive cycles of two sympatric skate species, Malacoraja senta and Amblyraja radiata, in the western Gulf of Maine

The smooth skate, Malacoraja senta, and thorny skate, Amblyraja radiata, are two commercially exploited batoids found within the Gulf of Maine. During the past five years, we conducted a large study to accurately describe important biological life history parameters previously lacking for these species. As part of that project, the current study reports our findings on the hormonal profiles associated with the reproductive cycles of M. senta and A. radiata. Blood samples were obtained from mature M. senta and A. radiata of both sexes from all months of the year, and plasma testosterone (T), estradiol (E₂) and progesterone (P₄) concentrations were determined using radioimmunoassay (RIA). In female M. senta and A. radiata, monthly T concentrations ranged from 4,522 pg ml⁻¹ to 1,373 pg ml⁻¹ and 31,940 pg ml⁻¹ to 14,428 pg ml⁻¹, E₂ concentrations from 831 pg ml⁻¹ to 60 pg ml⁻¹ and 8,515 pg ml⁻¹ to 2,902 pg ml⁻¹, and P₄ concentrations from 3,027 pg ml⁻¹ to 20 pg ml⁻¹ and 3,264 pg ml⁻¹ to 331 pg ml⁻¹, respectively. No statistical differences were detected between any months for any hormone. Estradiol concentrations were not correlated with ovary weight, shell gland weight, or diameter of the largest follicles in either species. Monthly T concentrations in male M. senta and A. radiata ranged from 23,146 to 12,660 pg ml⁻¹ and from 57,500pg ml⁻¹ to 24,737 pg ml⁻¹, while E₂ concentrations ranged from 7.5 pg ml⁻¹ to undetectable and 103 to 30 pg ml⁻¹, respectively. No statistical differences were observed between months for either steroid. Testosterone concentrations were weakly correlated with testes weight and percent of stage VI spermatocysts in A. radiata, however, no correlation was detected between T and stage VI spermatocysts in M. senta. Collectively, these data support the previous conclusion that M. senta and A. radiata of both sexes are capable of reproducing year round in the western Gulf of Maine.     

Differential Effects of the KitW Mutation with Other Kit Allelic Combinations on Male Sterility Among Inter- or Intrasubspecific Hybrids in Mice

The combination of the Kit ^W mutation and Kit ^S allele from Mus spretus leads to male hybrid sterility. The effects of other combinations between Kit ^W and Kit ^M from Mus m. molossinus or Kit ^N from Mus m. musculus on male reproductive ability were examined in this study. The Kit ^W/Kit ^M and Kit ^W/Kit ^N males were fertile and showed the normal pattern of spermatogenesis in most seminiferous tubules. There were two amino acid substitutions in the protein deduced from the cDNA sequence coded by the Kit ^M allele sequence and three in the Kit ^M allele compared with the protein from the + Kit allele of C57BL mice. These amino acid exchanges had no effect on the fertility of Kit ^W/Kit ^M and Kit ^W/Kit ^N males. Therefore, comparing the sequence data from cDNA coded by Kit ^M and Kit ^N alleles with that for the Kit ^S allele, we concluded that one or more amino acid exchanges in the extracellular domain would be the cause of male hybrid sterility in the Kit ^W/Kit ^S combination; these substitutions are Phe to Ser at position 72, Thr to Ala at 95, Ser to Arg at 101, Leu to Pro at 123, and Ile to Met at 1303     

Investigation of native isolates of entomopathogenic fungi for the biological control of three citrus pests

The pathogenicity of 15 isolates of Beauveria bassiana (Balsamo) Vuillemin, five isolates of Metarhizium anisopliae (Metschnikoff) Sorokin and one isolate of M. flavoviride (Gams and Rozsypal) were tested under laboratory conditions against the subterranean life stages of the citrus pests, Ceratitis rosa Karsch, C. capitata Wiedemann (Diptera: Tephritidae) and Thaumatotibia leucotreta Meyrick. (Lepidoptera: Tortricidae). When these citrus pests were treated with a concentration of 1×10⁷ conidia mL^?1, fungal isolates had a significantly greater effect on the adults of C. rosa and C. capitata than they did on the puparia of these two fruit fly species. Further, C. rosa and C. capitata did not differ significantly in their response to entomopathogenic fungi when adult and pupal mycosis were considered. Depending on fungal isolate, the percentage of T. leucotreta adults which emerged from fungal treated sand ranged from 5 to 60% and the percentage of pupae with visible signs of mycosis ranged from 21 to 93%. The relative virulence of the four most promising fungal isolates, as well as the commercially available B. bassiana product, BroadBand® (Biological Control Products, South Africa), were compared against one another as log-probit regressions of mortality against T. leucotreta which exhibited a dose-dependent response. The estimated LC₅₀ values of the three most virulent B. bassiana isolates ranged from 6.8×10⁵ to 2.1×10⁶ conidia mL^?1, while those of the least pathogenic ranged from 1.6×10⁷ to 3.7×10⁷ conidia mL^?1.     

Detection of three virulence genes alt, ahp and aerA in Aeromonas hydrophila and their relationship with actual virulence to zebrafish

Aims: To evaluate the frequency of the aerolysin (aerA), cytotoxic enterotoxin (alt) and serine protease (ahp) genes in Aeromonas hydrophila isolates from different sources, and to determine the relationship between the presence of these genes and virulence of A. hydrophila in zebrafish. Methods and Results: Aeromonas hydrophila isolates from clinical cases (n = 40), from healthy fish (n = 22) and from water environment (n = 21) were analysed with respect to the prevalence of aerA, alt and ahp genes by PCR assay. These virulence factors occur among clinical isolates as well as among isolates from healthy fish and water environment. The majority (97·6%) of the strains examined carried one or more virulence genes. The isolates were divided into seven genetic profiles on the basis of PCR result: aerA⁺alt⁺ahp⁺ (62·7%), aerA⁺alt⁺ahp^? (13·3%), aerA⁺alt^?ahp⁺ (10·8%), aerA^?alt⁺ahp⁺ (4·8%), aerA^?alt^?ahp⁺ (3·6%), aerA⁺alt^?ahp^? (2·4%) and aerA^?alt^?ahp^? (2·4%). A higher frequency of genetic group aerA⁺alt⁺ahp⁺ was determined in the isolates from diseased animals compared to those from healthy fish or water environments. Virulence properties of 26 representative strains belonging to the seven genetic profiles were further characterized. Results demonstrated that as the present of virulence genes increased, the proteolytic, haemolytic and cytotoxic activities of extracellular products also increased. And the 50% lethal doses (LD₅₀s) of aerA⁺alt⁺ahp⁺ isolates (<10⁵) in zebrafish were lower when compared with the strains expressing one or combinations of two virulence genes (>10⁶). Conclusions: Virulence properties of A. hydrophila correlated well with the presence of virulence genes tested. aerA⁺alt⁺ahp⁺ was more frequent virulence genotype in A. hydrophila isolates from clinical diseases than from healthy fish and water environment, and the aerA⁺alt⁺ahp⁺ isolates were more virulent to zebrafish compared to the other six genetic profiles. Significant and Impact of the Study: The detection for aerA, alt and ahp can be used for virulence typing of A. hydrophila isolates.     

Molecular mass and chain conformations of Rhizoma Panacis Japonici polysaccharides

The molecular mass and size of five water-soluble polysaccharides isolated from Rhizoma Panacis Japonici (RPJ) were determined with laser light scattering (LLS), size-exclusion chromatography (SEC) combined with LLS (SEC–LLS), dynamic light scattering (DLS), as well as transmission electron microscope (TEM). Their weight-average molecular masses (M_w) were 3.5 × 10⁴, 1.47 × 10⁵, 1.24 × 10⁶, 9.26 × 10⁵ and 1.36 × 10⁶, radii of gyration (<s²>_z^1/2) were 14.7, 31.7, 50.8, 41.8 and 40.4 nm, and hydrodynamic radii (R_h) were 19.9, 37.5, 66.2, 52.1, and 55.2 nm, respectively. The results showed that molecular masses and sizes of the polysaccharides were influenced by the pH and temperature of the extraction mediums. The conformation parameters were calculated from the above data according to polymer solution theory. The values of ρ (= <s²>_z^1/2/R_h) were from 0.7 to 0.8, exponents (ν) of <s²>_z^1/2 = k M_w^ν were from 0.31 to 0.43, and fractal dimension (d_f) were from 2.3 to 3.2, respectively. The results revealed that all of the polysaccharides existed as spheres in 0.15 M NaCl aqueous solution. TEM and atomic force microscope (AFM) further confirmed the spherical morphologies of these molecules. The spherical conformations of the polysaccharides were a result of their highly branched structures.     

Precursor of mast cells fixed in the skin of mice

The presence and origin of mast-cell precursors fixed in the skin tissue of mice were investigated. Giant granules of beige (C57BL/6-bg^j/bg^j, Chediak-Higashi syndrome) mice were used to distinguish different populations of mast cells. Pieces of the skin were grafted from the intact WBB6F¹ (WB × C57BL/6)F¹?+/+ mice onto the back of the WBB6F¹?+/+ mice which had been irradiated and injected with bone marrow cells of C57BL/6-bg^j/bj^j mice (bg^j/bg^j ?+/+ chimeras). Although the number of mast cells in the skin grafts decreased after the transplantation, the mast-cell precursors circulating in the bloodstream of bg^j/bg^j ?+/+ chimeras (bg^j/bg^j type) did not seem to enter into the skin grafts, because most of mast cells were of +/+ type after the recovery of mast-cell number to pregrafting levels. As a considerable proportion of +/+-type mast cells was labeled with ³H-thymidine, the recovery of mast-cell number in the grafts was attributed to the proliferation and differentiation of +/+-type precursor cells fixed in the skin tissue of the donor. On the other hand, the skin of WBB6F¹-W/W^v mice seemed to be depleted of fixed precursors, because most of mast cells were of bg^j/bg^j type in skin grafted from WBB6F¹-W/W^v mice to bg^j/bg^j?+/+ chimeras. Since the fixed precursor cells which proliferate and differentiate into mast cells after skin grafting may be transferred to WBB6F¹-W/W^v mice by bone marrow transplantation, such precursor cells seem to have been derived from the bone marrow.     

Biochemical genetics of TL antigens

TL antigens are class I glycoproteins which are expressed on thymocytes and which are coded by the Tla region of the major histocompatibility complex of the mouse. Biochemical analysis of TL molecules from different strains of mice revealed structural variation determined by the Tla region which is detectable by peptide mapping, isoelectric focusing, sodium dodecyl sulfate-polyacrylamide gel electrophoresis, two-dimensional gels, and by differential reactivity of allelic forms of TL molecules with a panel of anti TL reagents. The quantity of TL expressed on thymocytes is also influenced by the Tla region; three quantitative phenotypes were identified: high (Tla ^a, Tla ^d, Tla ^c), intermediate (Tla ^c, Tla ^f), and low (Tla ^b). (Relative amounts: 1000 : 100 : 1.) Some thymic leukemias arising in (Tla ^b, Tla ^c, Tla ^c) mice with genetically determined reduced levels of thymic TL were found to express TL molecules which were structurally indistinguishable from TL isolated from thymocytes but were present in larger amounts. This suggests that TL structural genes are intrinsically capable of full expression in all mice but that the Tla region of mice expressing an intermediate or low quantity of TL is marked by some feature which causes the thymocyte to express less than the full amount of TL possible.