Infectivity, growth, development and pathology of Echinostoma caproni (Trematoda) in the golden hamster

Laboratory hamsters (Mesocricetus auratus) were experimentally infected with 75 ± 15 metacercarial cysts of Echinostoma caproni. Worms were recovered from days 7 to 89 post-infection with eight to 90 (average 37) parasites in the small intestine. Worm wet weights averaged 0.85 mg at 10 days, 1.8 mg at 17 days, 3.4 mg at 45 days, and 7.7 mg at 89 days; average dry weights for the identical days were, 0.15, 0.30, 0.70 and 2.2 mg, respectively. The average body area of worms fixed in hot (80°C) alcohol-formalin-acetic acid was 0.21 mm² on day 3, 4.9 mm² on day 10, and 17.7 mm² on day 42. Clinical signs in some hamsters included progressive unthriftiness and watery diarrhea. Gross examination revealed enlarged lymphatic nodules along the length of the small intestine. The histopathological responses of hamsters to the parasite showed erosion of the intestinal villi with lymphocytic infiltration being the primary response; hemorrhagic areas were also observed in the villi.     

Infectivity, growth, and development of Echinostoma revolutum (Digenea: Echinostomatidae) in the golden hamster, Mesocricetus auratus

All 30 female golden hamsters, Mesocricetus auratus, each fed 100 +/- 25 metacercarial cysts of Echinostoma revolutum were found to be infected 2 to 105 days post-infection with 3 to 103 (avg. 38) flukes in the small intestine. Worm wet weights averaged 0.6 mg at 9 days, 3.5 mg at 14 days, and 19 mg at 42 days; average dry weights for the identical days were 0.2, 1.2 and 5.8 mg, respectively. The average body length of worms fixed in hot (80 C) alcohol-formalin-acetic acid was 0.33 mm on day 2, 5.11 mm on day 10, 9.30 mm on day 42 and 8.56 mm on day 105. Body and gonadal area increased rapidly from about day 5 to 15 and then less rapidly. Eggs of E. revolutum were seen in the feces of 10% of the hamsters by day 9, 89% by day 10 and 100% by day 11. Eggs teased from worms, embryonated in tap water, and produced miracidia which infected lab-reared Helisoma trivolvis.     

Evaluation of the Entomopathogenic Fungi Beauveria bassiana and Paecilomyces fumosoroseus for Microbial Control of the Silverleaf Whitefly, Bemisia argentifolii

Collaborative research was conducted at the USDA-ARS Subtropical Agricultural Research Center in southern Texas to assess the microbial control potential of Beauveria bassiana and Paecilomyces fumosoroseus against Bemisia whiteflies. Laboratory assays demonstrated the capacity of both pathogens to infect Bemisia argentifolii nymphs on excised hibiscus leaves incubated at relative humidities as low as 25% at 23 ± 2°C (ca. 35% infection by B. bassiana and P. fumosoroseus resulted from applications of 0.6–1.4 × 10³ conidia/mm² of leaf surface). In small-scale field trials using portable air-assist sprayers, applications at a high rate of 5 × 10¹³ conidia in 180 liters water/ha produced conidial densities of ca. 1–2.5 × 10³ conidia/mm² on the lower surfaces of cucurbit leaves. Multiple applications of one isolate of P. fumosoroseus and four isolates of B. bassiana made at this rate at 4- to 5-day intervals provided >90% control of large (third- and fourth-instar) nymphs on cucumbers and cantaloupe melons. The same rate applied at 7-day intervals also provided >90% control in zucchini squash, and a one-fourth rate (1.25 × 10¹³ conidia/ha) applied at 4- to 5-day intervals reduced numbers of large nymphs by >85% in cantaloupe melons. In contrast to the high efficacy of the fungal applications against nymphs, effects against adult whiteflies were minimal. The results indicated that both B. bassiana and P. fumosoroseus have strong potential for microbial control of nymphal whiteflies infesting cucurbit crops.     

Increased resistance of splenectomized mice to Sporothrix schenckii infection

The susceptibility of splenectomized mice to Sporothrix schenckii was studied, and the role of the spleen in the host defense is discussed. S. schenckii Sp-1 and ddy male mice were used. The mice were divided into 3 groups consisting of splenectomized, sham-operated and intact mice. Each mouse was inoculated intravenously with 2×10⁶ yeast cells 7 days after operation and the mice were sacrified at adequate intervals for 30 days. Then histological sections stained with H&E or by PAS were prepared from various visceral organs. Using the liver sections the number of yeast cells in a 40 mm² was counted. Furthermore, the colony forming unit in 100 mg of the liver tissue was compared to each other.In the sham-operated and intact mice many purulent lesions appeared on the 5th day. On the 8th day mononuclear cells accumulated at the foci, and on the 10th day most of the foci became granulomatous. The number of yeast cells in granulomatous lesions reached a peak on the 10th day and thereafter decreased abruptly. On the other hand, in the splenectomized mice approximately half of foci became granulomatous on the 5th day, and the number of yeast cells in the foci began to decrease after the 5th day.There were definite differences in the colony forming unit between the splenectomized and sham-operated or intact mice sacrificed 9 days after inoculation. The colony forming unit of the former is 9.3×10⁵ on the average, while that of the latter two is 5.6×10⁶ and 5.1×10⁶ on the average, respectively.In conclusion the resistance of ddy mice to S. Schenckii infection is enhanced due to splenectomy.     

Life table demography and population growth of Brachionus variabilis Hempel, 1896 in relation to Chlorella vulgaris densities

We studied the life history variables and population growth characteristics of Brachionus variabilis, which was recorded for the first time from Mexico. The animals were fed Chlorella, using five concentrations (0.25, 0.5, 1, 2 and 4 × 10⁶ cells ml^–1) at 25 °C. Food density was observed to have significant effect on life expectancy, average lifespan, gross reproductive rate, net reproductive rate, generation time and population growth rate. The average lifespan ranged from 3 to 6 days depending on the food density. The net reproductive rate ranged from 2 to 7 neonates female^–1 d^–1. The rate of population increase per day varied from 0.14 to 0.35. The highest net reproductive rate and average lifespan and life expectancy were recorded at Chlorella concentrations of 1 × 10⁶ and 2 × 10⁶ cells ml^–1.     

Glycosphingolipids in feces of germ-free rats as a source for studies of developmental changes of intestinal epithelial cell surface carbohydrates

Non-acid and acid glycosphingolipids were isolated from feces of one litter of germ-free rats from day 17 to day 51. Quantitative and qualitative changes described for small intestine of conventional rats [Bouhours D, Bouhours J-F (1981) Biochem Biophys Res Commun 99:1384–89] were also found in the feces of these germ-free rats. A decrease in lactosylceramide and sialyllactosylceramide excretion and a change fromN-acetylneuraminic acid toN-glycoloylneuraminic acid, as well as an appearance of type 1 chain blood group H-active penta- and decaglycosylceramides were observed during the weaning period. Thus the dramatic changes seen in rat intestinal glycosphingolipids postnatally seem to be primarily regulated by non-microbial factors.Abbreviations G_M3 G_M3-ganglioside, II³NeuAc-LacCer or II³NeuGc-LacCer - SPG IV³NeuAc-nLcOse₄Cer - G_M1 G_M1-ganglioside, II³NeuAc-GgOse₄Cer     

Effect of Temperature on the Life History of Cirrospilus sp. near lyncus (Hymenoptera: Eulophidae), a Parasitoid of Phyllocnistis citrella (Lepidoptera: Gracillariidae)

The effect of four constant temperatures on the life history of Cirrospilus sp. near lyncus was examined in the laboratory. This species is one of the most abundant generalist indigenous parasitoids of the citrus leafminer, Phyllocnistis citrella Stainton, in Spain. Adult lifespan of C. sp. near lyncus decreased from 50.2 to 9.1 days as temperatures increased from 15 to 30°C, respectively. Both gross fecundity and host-feeding were highest at 20°C (170.48 eggs and 32.33 hosts). Oviposition rates were optimal at higher temperatures (5.22 eggs per day at 25°C and 4.79 eggs per day at 30°C) and were dependent on female age. In contrast, host-feeding rates for a given temperature did not depend on age. Generation time decreased with increasing temperatures from 68.05 days at 15°C to 12.19 days at 30°C. Net reproduction peaked at 20°C (68.86 viable females per female). Intrinsic rate of increase doubled from 15°C (0.059 females per female per day) to 20°C (0.127 females per female per day) and almost doubled again from 20 to 30°C (0.210 females per female per day). Given these parameters, C. sp. near lyncus could perform optimally in the area occupied by P. citrella in the Mediterranean region.     

Growth characters and photosynthetic capacity of <Emphasis Type="BoldItalic">Gracilaria lemaneiformis</Emphasis> as a biofilter in a shellfish farming area in Sanggou Bay,China

Experiments on growth characters and ecological functions of the macroalgae Gracilaria lemaneiformis, collected from south China, were conducted in polyculture areas of kelp and filter-feeding bivalve in Sanggou Bay in Weihai City, Shandong, in north China from May 2002 to May 2003. The results of 116 days cultivation showed that the average wet weight of alga increased 89 times from 0.1 to 8.9 kg rope⁻¹, with an average specific growth rate (based on wet weight) of 3.95% per day. The most favorable water layer for its growth was 1.0–1.8 m below the surface in July and August, with an average specific growth rate of 8.2% per day in 30-day experiments. Photosynthetic activity changed seasonally, with an average of 7.3 mg O₂ g dw⁻¹ h⁻¹. The maximum rate (14.4 mg O₂ g dw⁻¹ h⁻¹) was recorded in July, or 19.3 mg CO₂ g dw⁻¹ h⁻¹, while the minimum (0.40 mg CO₂ g dw⁻¹ h⁻¹) was in April. This study indicated that the culture of G. lemaneiformis is an effective way to improve water quality where scallops are cultivated intensively.     

Purification and some properties of a starch debranching enzyme ofHendersonula toruloidea

An extracellular, debranching isoamylase fromHendersonula toruloidea ATCC 64930, grown on starch, was purified 12-fold to an electrophoretically homogeneous state. The purified enzyme (estimated mol wt 83000) was optimally active at pH 6.0 and 50°C and remained active when held at 70°C (30 min) and at pH 6 to 8 for 24 h. Na⁺, Fe²⁺ and Ba²⁺ (at 5mm) enhanced enzyme activity while Hg²⁺, Zn²⁺ and Cu²⁺ (at 5mm) were inhibitory. The enzyme hydrolysed amylopectin (Km, 0.25 mg/ml), forming maltose, maltotriose and maltotetraose and hydrolyzed glycogen (Km, 0.29 mg/ml) and soluble starch (Km, 0.42 mg/ml) forming maltotriose and maltotetraose. Pullulan was not hydrolyzed.     

Effects of temperature, salinity and irradiance on the growth of the red tide dinoflagellate Gyrodinium instriatum Freudenthal et Lee

The effects of temperature, salinity and irradiance on the growth of the red tide dinoflagellate Gyrodinium instriatum Freudenthal et Lee were examined in the laboratory. Exposed to 45 different combinations of temperature (10–30 °C) and salinity (0–40) under saturating irradiance, G. instriatum exhibited its maximum growth rate of 0.7 divisions/day at a combination of 25 °C and a salinity of 30. Optimum growth rates (>0.5 divisions/day) were observed at temperatures ranging from 20 to 30 °C and at salinities from 10 to 35. The organism could not grow at ≤10 °C. In addition, G. instriatum burst at a salinity of 0 at all temperatures, but grew at a salinity of 5 at temperatures between 20 and 25 °C. It is noteworthy that G. instriatum is a euryhaline organism that can live under extremely low salinity. Factorial analysis revealed that the contributions of temperature and salinity to its growth of the organism were almost equal. The irradiance at the light compensation point (I₀) was 10.6 μmol/(m² s) and the saturated irradiance for growth (I_s) was 70 μmol/(m² s), which was lower than I_s for several other harmful dinoflagellates (90–110 μmol/(m² s)).     

The response of intact Strongyloides ratti infective (L3) larvae to substrates and inhibitors of respiratory electron transport

Live, intact third-stage larvae (L3s) of Strongyloides ratti in the absence of exogenous substrates consumed oxygen at a rate (E-QO₂) of 181.8 ± 12.4 ng atoms min⁻¹ mg dry weight⁻¹ at 35°C. Respiratory electron transport (RET) Complex I inhibitor rotenone (2 μm) produced 33 ± 6.5% inhibition of the E-QO₂. Unusually the rotenone-induced inhibition was not relieved by 5 μm-succinate. The E-QO₂ of intact L3s was refractory to RET Complex III inhibitor antimycin A at 2 μm; 4 μm-antimycin inhibited ≤ 10% of the E-QO₂. The electron donor couple ascorbate/TMPD augmented the E-QO₂ in the presence of rotenone (2 μm) and antimycin A (4 μm) by 110%. Azide (1 mm) stimulated the antimycin A refractory QO₂ by 36.6 ± 7.2% which was only partially inhibited by 1.0 mm-KCN (IC₅₀ = 0.8 mm). The data suggest the presence of classical (CPW) and alternate (APW) electron transport pathways in S. ratti L3s.     

The effect of host age ofLymantria dispar larvae [Lep.: Lymantriidae] on the development ofGlyptapanteles liparidis [Hym.: Braconidae]

The endoparasitic development ofG. liparidis was examined in 3 different host stages of gypsy moth larvae. Hatching ofG. liparidis-larvae occurred 3 to 5 days after oviposition in hosts parasitized during their premoulting period, and after 5 to 7 days in those parasitized in the 3^rd midinstar state. The parasites generally moulted to the 2^nd larval instar between the 11^th and 13^th day in the first group, and between the 13^th and 15^th day in the latter, when they had reached a volume of 0.04–0.05 mm³. The positive correlation between host ecdysis and the ecdysis of 1^st stadium larvae to L₂ suggested that host moulting influenced the development of the parasitoid larvae. Emergence from the host larvae occurred at 20°C after 27 days on average, and coincided with the parasites moulting to the 3^rd instar. Five to 7 days after spinning their cocoons near the developmentally arrested host larva, the male, and 1 to 2 days later the female wasps eclosed. Due to the variation in the number of parasites per host, no difference was observed between the hosts parasitized at various stages; however, a tendency for later parasitized hosts to contain more parasite larvae was evident. The nutritional conditions of the moth parental generation influenced both host and parasite development. On the other hand no influence of host age was observed on emergence dates of larvae and wasps.      

Biomass relationship to growth and phosphate uptake ofPseudomonas fluorescens,Escherichia coli andAcinetobacter radioresistens in mixed liquor medium

The ability ofPseudomonas fluorescens, Escherichia coli andAcinetobacter radioresistenns to remove phosphate during growth was related to the initial biomass as well as to growth stages and bacterial species. Phosphate was removed by these bacteria under favourable conditions as well as under unfavourable conditions of growth. Experiments showed a relationship between a high initial cell density and phosphate uptake. More phosphate was released than removed when low initial cell densities (10²–10⁵ cells ml^–1) were used. At a high initial biomass concentration (10⁸ cells ml^–1), phosphate was removed during the lag phase and during logarthmic growth byP. fluorescens. Escherichia coli. at high initial biomass concentrations (10⁷ cells ml^–1), accumulated most of the phosphate during the first hour of the lag phase and/or during logarithmic growth and in some cases removed a small quantily of phosphate during the stationary growth phase.Acinetobacter radioresistens, at high initial cell densities (10⁶, 10⁷ cells ml^–1) removed most of phosphate during the first hour of the lag phase and some phosphate during the stationary growth phase.Pseudomonas fluorescens removed phosphate more thanA. radioresistens andE. coli with specific average ranges from 3.00–28.50 mg L^–1 compared to average ranges of 4.92–17.14 mg L^–1 forA. radioresistens and to average ranges of 0.50–8.50 mg L^–1 forE. coli.     

Studies on mixed mass cultivation of Anabaena spp. (nitrogen-fixing blue-green algae, cyanobacteria) on a large scale

Studies on mixed mass cultivation of Anabaena spp. on a large scale (5170m²) were conducted continuously for 3 years. Under the continental monsoon climate in northern subtropics (30°N, 115°E), 7–11 g dry weight m⁻² day⁻¹ of microalgal biomass on average was harvested in simple plastic greenhouses in the effective growth days during the warmer seasons. The maximum productivity was 22 g m⁻² day⁻¹ in the middle of summer. Observations on the productive properties of strains of Anabaena spp. indicated that they were different from and could compensate for each other in their productivities and adaptations to the seasonal changes. With different lining materials (PVC sheets, concrete, sand and soil) in the culture ponds, no significant variation of productivity was found, but bubbling with biogas in the middle of the day and the application of some growth regulating substances (2,4-D, NaHSO₃ and extracts of oyster mushroom spawn) was able to improve the production. The cost of microalgal biomass in this way was around 0·75–1·0 US dollar(s) per kilogram.     

Impact of Enterococcus faecium on specific activity of disaccharidases in small intestine of gnotobiotic pigs

The aim of the present work was to study the changes in the activity of disaccharidase enzymes (lactase. maltase, saccharase) in the small intestine of gnotobiotic pigs aged 0–35 days and inoculated with Enterococcus faecium. The continual decrease of lactase activity was observed from the 14^th day of age up to the end of the experiment. The most significant decrease of specific lactase activity in the duodenum (2.1 μmol/mg protein/hour) was noted from the 21^st to the 28^th day of age. On the other hand, the specific saccharase activity increased moderately during the post weaning period and maltase activity maintained a constant level. Presented at the Second Probiotic Conference, Košice, 15–19 September 2004, Slovakia.     

Infectivity of two members of theEntomophthora muscae complex [Zygomycetes: Entomophthorales] forMusca domestica [Dipt.: Muscidae]

Dose-mortality studies were conducted with 2 members of theEntomophthora muscae (Cohn) Fresenius complex from southern California (CA) and Denmark (DA) infecting house flies,Musca domestica L., from southern California. Primary conidia of the DA form were significantly more infective (LC₅₀=34 conidia/mm²) than primary conidia of the CA form (LC₅₀=67 conidia/mm²) for 2–7 days old (‘young’) flies. Infectivity (single experimental trial) for 10–14 day old (‘old’) flies of primary conidia of the CA form (LC₅₀=34 conidia/mm²) was similar to that of the DA form for young flies. Secondary conidia of the CA form were markedly more infective (LC₅₀=0.36 conidia/mm²) than primary conidia of either form. Dose had no significant effect on incubation period for primary conidia of either form, but increased doses resulted in significantly shorter incubation periods for flies exposed to secondary conidia of the CA form.      

Water-soluble granules containing <Emphasis Type="Italic">Bacillus megaterium</Emphasis> for biological control of rice sheath blight: formulation,bacterial viability and efficacy testing

Endospores of B. megaterium were formulated in granule formulations with sodium alginate, lactose and polyvinylpyrrolidone (PVP K-30) by the wet granulation technique. The granule formulation exhibited good physical characteristics, such as high-water solubility and optimal viscosity, that would be suitable for spray application. The bacteria remained viable in the dry granule formulation at 10⁹ c.f.u./g after 24 months storage at room temperature. Under laboratory conditions, aqueous solutions of the formulation showed high activity against mycelial growth of R. solani (99.64 ± 0.14% mycelial inhibition). High viability of the bacterial antagonist on leaf sheath and leaf blade at day 7 after spraying with the formulation was observed (approximately 10⁶ c.f.u./g of plant). Application of an equivalent number of un-formulated endospores resulted in much loss of the bacterial endospores even 1 day after application. In a small pilot field study, an aqueous solution of the formulation (3%w/v) applied by spraying at days 1, 5 and 10 after pathogen inoculation of the rice plants was more effective in suppressing rice sheath blight disease than one application of a fungicide (Iprodione) at day 1. Additionally, rice plants sprayed with the aqueous solution of the granule formulation had higher panicle and whole kernel weights than those of fungicide-treated and control (untreated) plants.     

The lethal effects of gamma irradiation on larvae of the Huhu beetle, Prionoplus reticularis: a potential quarantine treatment for New Zealand export pine trees

Gamma irradiation was investigated as a possible method for disinfestation of huhu beetle larvae, Prionoplus reticularis White, in Pinus radiataD. Don. Larvae of four representative size classes were irradiated at six doses, and the lethal dose (LD₉₉) calculated from mortality data 3 days and 10 days post treatment. All larval size classes showed a similar sensitivity to gamma irradiation and required 3677 Gray (Gy) and 2476 Gy for a LD₉₉ 3 and 10 days post-treatment, respectively. The penetration of gamma irradiation into pine wood was found to be lowest in freshly cut logs, and decreased linearly at a rate of 0.698 Gy mm^–1 of wood. The penetration was greatest in wood that had been stored for 2 years, and decreased 0.512 Gy mm^–1 of wood. These results are likely to be correlated with wood moisture content. Gamma irradiation appears to be a potential alternative method to fumigation for quarantine treatment of P. reticularis.     

Pseudomonas aeruginosa,a facultative pathogen of red palm weevil,Rhynchophorus ferrugineus

Pseudomonas aeruginosa was identified as a facultative pathogen of red palm weevil. Intra-haemocoelic injection of the pathogen within larvae and pre-pupae was more effective at killing the insects [with a median lethal dose (LD₅₀) of 9×10² to 2×10³ bacteria/insect] than inoculation by force feeding (LD₅₀ of 10⁵ to 4×10⁵ bacteria/insect) or by wading the insects in a suspension of the pathogen (LD₅₀ of 10⁵ to 2×10⁵ bacteria/insect). Injection of 3×10³ bacteria/insect killed 69% of larvae; small larvae were more susceptible (LD₅₀ of 9×10⁵ bacteria/larva) than either larger larvae (LD₅₀ of 10³ bacteria/larva) or pre-pupa. The median time to death of the small larvae following injection of P. aeruginosa was about 6 days but that following force feeding or wading was about 8 days. A secondary invader, Serratia marcescens, had no effect on the pathogenicity of P. aeruginosa but hastened death of larvae by about 3 days.A. Banerjee and T.K. Dangar were with the Central Plantation Crops Research Institute, Regional Station, Kayangulam 690 533, Kerala, India. They are now with the Central Rice Research Institute. Cuttack 753 006, Orissa, IndiaCPCRI research paper no. 870.     

Seasonal changes in composition and biomass of epiphytic algae on the macrophyte Ranunculus penicillatus in a chalk stream,with estimates of production,and observations on the epiphytes of Cladophora glomerata

The epiphytic algae on surfaces of the macrophyte Ranunculus penicillatus (Dumort.) Bab. var. calcareus (R. W. Butcher) C. D. K. Cook and on Cladophora glomerata (L.) Kütz growing in the River Itchen at Otterbourne near Southampton were studied between February 1984 and June 1985. The river at this site has a mean flow rate of 0.33 m s^–1, and is about 16 m wide and on average 20 cm deep, with a discharge ranging through the year between 0.34 and 2.46 m³ s^–1. The pH of the river varies little around 8.2, with a mean alkalinity of 236 mg HCO _inf3 ^sup1 l^–1, because of its origin from chalk springs. Ranunculus grows throughout the year, with peaks of biomass in spring and autumn. It forms a very large surface for attachment of epiphytes, and covers on average 40% of the stream bed. Numbers of epiphyte cells removed from Ranunculus ranged through the year between 52 × 10³ and 271 × 10³ cells mm^–2 stream floor, with maximum numbers in April, and a secondary peak in October. This pattern partly reflects fluctuations in the biomass of Ranunculus; the number of cells per unit area of plant surface showed a broader spring peak and lower fluctuations in other seasons. Diatoms formed 65 to 98% of these epiphyte cells, with chlorophytes reaching their peak (10%) in summer and cyanophytes (25%) in autumn. Estimates of biomass of these epiphytic forms, derived from measurements of chlorophyll c, indicate a range between 30 and 100 g dry weight m^–2 of weed bed. Colonisation studies showed that the algae grow and reproduce throughout the year, with a mean generation time of about 5 days, suggesting an annual production of about 3 kg dry weight m^–2 of weed bed, which makes epiphytic algae the principal primary producers in the stream. The numbers and biomass of epiphytic algae on Cladophora are considerably less.The species of epiphytic algae found on Ranunculus were generally similar to those growing on Cladophora, and to epilithic algae on pebbles of the stream bed, but different species were dominant on the different substrata. Algal cells in the water column were all derived from benthic habitats, although their relative abundance was very different.