Laboratory tests of fungicides against Peronospora parasitica (Pers. ex Fr.) Fr

Methods were investigated of culturing Peronospora parasitica on detached cotyledons in plastic box moist chambers in constant-temperature cabinets. Light was essential and a temperature of 15 °C was most suitable for infection and survival of cotyledons. Using such cotyledons the protectant and eradicant actions of various fungicides against P. parasitica were compared with those of zineb. Captafol, Daconil 2787, dichlofluanid and propineb were markedly superior to zineb as protectants, being highly effective at 0·025 % a.i. None of these compounds (at 0·2 % a.i.) checked fungal development when applied 5 h after inoculation, during which time the fungus had penetrated the cotyledons, and it was concluded that the fungicides had thus no internal effect. Internal fungistasis was shown, however, by a maneb-nickel sulphate fungicide and also by nickel sulphate. The latter (at 0·4% NiSO₄.6H₂O) gave some disease control when applied up to 72 h after inoculation and appeared to be translocated within the tissue of the cotyledon.     

Soil-applied fungicides for controlling take-all in field experiments with winter wheat

Soil treatment fungicides were tested against take-all (Gaeumannomyces graminis var. tritici) in three field experiments with winter wheat. Fungicides were applied as drenches either before sowing in autumn, and incorporated by rotary harrowing, or to the crop in spring. The most effective treatments were autumn applied benomyl (20 kg/ha) and nuarimol (0·55-4·4 kg/ha). However, the highest nuarimol concentration depressed yield. Benomyl sometimes induced a resurgence of take-all in the second wheat crop after treatment. Nuarimol had no adverse effects in subsequent crops, and neither fungicide hindered the onset of take-all decline in a third crop after treatment. The possible value of soil treatment in future control strategies is discussed.     

Chemical control of Alternaria brown spot on Minneola tangelo in South Africa

The effectiveness of several groups of fungicides was compared with that of copper and mancozeb for their ability to control Alternaria brown spot on Minneola tangelo. Variables used for comparing fungicides in order of importance were: performance index, the number of exportable fruits per tree, the number of fruits per tree free of Alternaria lesions and total number of fruits per tree. Percentage exportable fruits was not a reliable variable for comparing treatment efficiency. During the 1992/1993 evaluations iprodione, difenoconazole, and procymidone showed promising control of the disease. Other than for difenoconazole, members of the triazole group were not effective, especially at dosages below 10 g a.i. 100 litre ^] water. Moderate control of Alternaria brown spot was obtained using copper and mancozeb. During the 1993/1994 season spray programmes with copper oxychloride, mancozeb, maneb plus zinc oxide, procymidone, iprodione, and tebuconazole were compared in field trials, for the control of Alternaria brown spot on Minneola tangelo. Mancozeb applied at 2-weekly intervals was the most effective spray programme, followed in order of efficiency by programmes using procymidone, maneb plus zinc oxide, and mancozeb applied at 4- and at 3-weekly intervals. Iprodione and tebuconazole were not particularly effective in this evaluation, although they performed significantly better than the control treatment. Trees treated with copper oxychloride produced poorly, in spite of a high percentage exportable fruits.     

A Cucumber Disease Caused by Alternaria alternata and its Control

A severe leaf spot disease of cucumber caused by a pathotype of Alternaria alternata (Fr.) Keissler was recorded recently in plastic houses in Crete. Lesions ranged in size of a pin point to over 5 cm in diameter, with necrotic tissue on most of their area and a surrounding yellow zone. The pathogen grew satisfactorily on PDA at temperatures between 5 °C–40 °C and spore germination occurred in the range less than 10 °C to over 37 °C. Optimum temperature in both cases was near 26 °C. Of,13 fungicides tested in vitro, sodium omadine, etem, dichlofluanid, captan and folpet were the most inhibitory on spore germination, and iprodione, sodium omadine and dichlofluanid on mycelial growth. Of 25 fungicides applied on two leaf cucumber plants 24 h before inoculation, maneb, etem, dichlofluanid and chlorothalonil were the most effective. When the last fungicides, plus mancozeb, were applied 24 h after inoculation only maneb was effective. In greenhouse experiments, iprodione, prochloraz-manganese-complex, chlorothalonil, dichlofluanid, guazatine, maneb and etem were the most effective for disease control, while mancozeb was less effective. The local cucumber cv. Knossos and the Dutch F₁ hybrids Evadan, Frella, Herta, Malfa, Mazourka, Pepinex 69 and Renova were all susceptible to infection.     

Effects of Field Application of Tebuconazole on Yield, Yield Components and the Mycotoxin Content of Fusarium-infected Wheat Grain

Winter wheat cultivar Basalt was artificially inoculated with Fusarium culmorum at the end of anthesis and treated with the systemic fungicide tebuconazole (Folicur^®) a few days before and/or after inoculation. Check plots remained uninoculated and unsprayed. Head infections, yield, yield components and the percentage of Fusarium‐ infected kernels were determined. Artificial Fusarium inoculation lowered yield significantly by 24.2‐45.0%. Any fungicide treatment saved yield, thousand grain weight and kernel numbers per head. Pre‐infectional application of tebuconazole was superior to application carried out post‐infection. Moreover, the fungicide controlled deoxynivalenol (DON) synthesis in the field to a considerable extent, and enabled good control of Fusarium head blight, glume blotch and the percentage of Fusarium‐infected kernels. The levels of Fusarium kernel infection after harvest clearly reflected the DON content of w heat grain.     

The effect of the weight of the seedling-derived tuber on subsequent clonal generations in a potato breeding programme

Cultures of Polymyxa graminis were maintained in roots of barley plants grown in sand at different temperatures using Wisconsin soil temperature tanks. At 17 – 20°C, the minimum time from inoculation with cystosori to the production of zoospores from the inoculated roots was 2 – 3 wk. At 11 – 20°C many zoospores were produced but the incubation period was longer at the lower temperatures. Above 20°C little fungal development occurred. The duration of motility of zoospores ranged from c. 1 h to > 24 h. Bovine serum albumen (BSA) prolonged motility but glycine and glucose had no effect or, at higher concentrations, were toxic. Zoospores were rapidly immobilised by zinc ions in solution at or above 10μg/ml. In some experiments BSA added to the zoospore suspension greatly increased transmission of barley yellow mosaic virus (BaYMV) while glucose, glycine and ovalbumen decreased it. When seedlings were incubated with zoospore suspensions for 24 h at different temperatures, BaYMV transmission was high (> 60%) at 10, 15 and 20°C but there was little at 5 or 25°C. In experiments to determine the time taken for zoospore penetration, seedlings were incubated in suspension for different periods of time and then rinsed in zinc sulphate solution to kill free zoospores. Between 3 and 3·5 h was needed for zoospores to establish infection. Transmission occurred equally to plants of various ages between 3 days and 7·5 wk.     

Studies on plant growth-regulating substances. XLII. Abscisic acid as a genetic character related to drought tolerance

A single fentinhydroxide spray at the end of May controlled Ramularia leaf spot in in-situ sugar-beet seed crops and increased seed yield on average by 4%. Sprays controlled leaf spots most effectively in the Cotswolds, but yield increases were larger in Lincolnshire. Benomyl controlled leaf spot as well as did fentinhydroxide; thiabendazole was less effective, and maneb and cuprous oxide gave poor control.     

Studies of the infection of the winged bean by Synchytrium psophocarpi in Papua New Guinea

Infection of leaves and stems of Psophocarpus tetragonolobus by Synchytrium psophocarpi only occurred following spray inoculation of motile zoospore suspensions and incubation for a minimum of 12 h in polyethylene bags or a mist chamber. The incubation period was 7 days and generation time 22 days at temperatures of 31 ^oC max, 24 ^oC min and r.h. of 90% max, 70% min. Young, 1–2 day-old leaves were most susceptible; there was no infection on 10 day-old leaves and susceptibility was not increased by the removal of leaf waxes. No infection occurred when plants were grown from seed from infected pods, seed inoculated with zoospores or sporangia and seed sown in soil containing infected leaf debris. Resting spores were not found in infected tissues stored for 12 wk or in plant debris. S. psophocarpi did not infect Arachis hypogaea, Glycine max, Phaseolus aureus, P. coccineus, P. vulgaris, Pisum sativum, Psophocarpus scandens, Vicia faba, Vigna sesquipedalis and V. unguiculata. S. minutum did not infect winged bean. Inoculation confirmed the susceptibility of the winged bean lines UPS 62, UPS 122, UPS 126 and resistance of two Thai winged bean lines 1602/1 and 1611/2.     

Tomato leaf mould: its assessment and effects on yield

When the natural incidence of Cladosporium fulvum Cooke, occurring at a relatively late stage of cropping, was controlled with fungicides, comparisons with unsprayed control plants showed that there was an interval of c. 6 weeks between the incidence of severe infection, when the fungus was colonizing at least 50 % of the host's leaf area, and fruit yield decreases. Eye estimates made on separate leaves usually over-estimated the percentage of infection. Correction factors were derived from the regression of eye estimates on infected areas calculated from leaf tracings. Whole-plant infection indices were obtained by averaging corrected values of percentage infection separately assessed on leaves immediately above each fruit truss. Although dichlofluanid and tank-mixed zineb decreased the incidence of C. fulvum more than captan sprays, tomato yields (including green fruit) were increased similarly by all three fungicides from 2·99 to an average of 3·51 kg/plant during c. 3 months' picking. Decreasing the intervals between successive sprays from 21 to 7 days greatly decreased leaf mould infection but did not significantly affect yields. Restricting applications to plant tops (namely foliage from the fourth-youngest truss to the mainstem apex) gave yield benefits equal to those gained from sprays applied to whole plants.     

In vitro assays on the susceptibility of four species of nematophagous fungi to anthelmintics and chemical fungicides/antifungal drug

Using nematophagous fungi for the biological control of animal parasitic nematodes will become one of the most promising strategies in the search for alternative chemical drugs. The purpose of this study was to check the in vitro activity of four anthelmintics, four chemical fungicides and two antifungal drugs on the spore germination of nematophagous fungi: Duddingtonia flagrans (SF170), Arthrobotrys oligospora (447), Arthrobotrys superba (435) and Arthrobotrys sp. (PS011). A modified 24-well cell culture plate assay was conducted to evaluate the susceptibility of nematophagous fungi against drugs tested by calculating the effective middle concentrations (EC₅₀) of each tested drug to inhibit the germination of fungal spores. EC₅₀ ranged between 0·7 and 47·2 μg ml⁻¹ for fenbendazole, thiabendazole and ivermectin, except levamisole (546·5–4057·8 μg ml⁻¹). EC₅₀ of tested fungicides was 0·6–2·3 μg ml⁻¹ for carbendazim, 55·9–247·4 μg ml⁻¹ for metalaxyl, 24·4–45·2 μg ml⁻¹ for difenoconazole, and 555·9–1438·3 μg ml⁻¹ for pentachloronitrobenzene (PCNB). EC₅₀ of two antifungal drugs was 0·03–3·4 μg ml⁻¹ for amphotericin B and 0·3–10·9 μg ml⁻¹ for ketoconazole. The results showed that 10 tested drugs, except for levamisole and PCNB, had in vitro inhibitory effects on nematophagous fungi. The chlamydospores of D. flagrans had the highest sensitivity to nine tested drugs, except for ketoconazole.     

Effects of Leaf Age, Inoculum Dose and Freezing on Development of Chocolate Spot (Botrytis fabae) Lesions on Field Bean (Vicia faba) Leaves

Botrytis fabae spore suspensions containing c. 1, 10, 10², 10³, 10⁴, 10⁵, or 10⁶ spores/ml were used to inoculate 5, 17 or 30-day-old field bean leaves. The percentages of the leaf areas covered by, chocolate spot lesions and the percentages of the leaf areas bearing conidiophores were assessed 1, 6, 12, 14, and 19 days after inoculation. The percentage of the area covered by lesions and the percentage of the area bearing conidiophores (logit-transformed) increased linearly with increasing spore concentration (log₁₀-transformed). The proportions of leaf areas covered by lesions and bearing conidiophores were both greater on 17 and 30-day-old leaves than on 5-day-old leaves. The rate of lesion growth increased with both increasing inoculum dose and increasing leaf age. Generally there was no interaction between the effects of leaf age and the effects of inoculum dose on either lesion growth or sporulation. Two days after inoculation with suspensions of either 10⁴ or 10⁶ spores/ml, 7-day-old leaves grown at 15°C were transferred to –16°C or 2.5°C or kept at 15°C for 4 days. Two days later more spores had been produced on leaves which had been frozen (–16°C) than on, leaves kept at 2.5°C.     

Application of chemical and biological agents for the management of frosty pod rot (Moniliophthora roreri) in Costa Rican cocoa (Theobroma cacao)

This article describes two field trials carried out at La Lola, Costa Rica, to assess control measures against frosty pod rot of cocoa (Theobroma cacao) caused by Moniliophthora (Crinipellis) roreri. In the first, factorial, trial the control agents were applied using motorised mistblowers (MMs) and hydraulic sprayers fitted with a narrow angle cone nozzle. There was an interaction between agents and application methods; together with previous application data for the most active fungicide (copper hydroxide), these trials indicate that best yields are achieved with sprays that maximise deposits on pods. We describe the droplet size spectra produced by a Stihl SR400 MM under a range of conditions because this has become the standard method of fungicide application in this series of trials at La Lola. The factor that had the largest effect on droplet size spectrum was the presence or the absence of a detachable baffle plate in front of the air‐shear nozzle. In both trials described here, MMs were fitted with baffle plates, a formulation pump and restrictor transmitting 550 mL min^?1 to deliver an estimated equivalent of 190 L ha^?1. Copper hydroxide as prophylactic applications at 1500 g a.i. ha^?1 have, to date, shown the most consistent (but incomplete) improvement in healthy pod yield. Use of copper fungicides may be cost effective when farm‐gate cocoa prices exceed approximately $1.25 kg^?1. In these trials, isolates of the hyperparasitic fungi Clonostachys byssicola and Trichoderma asperellum and two off‐patent triazole fungicides (bitertanol and triadimenol) made no significant improvement to healthy yields. The systemic oxathiin fungicide flutolanil, at a dosage of 300 g a.i. ha^?1, appears to protect pods substantially at early stages but gives proportionately less control of M. roreri than copper at later stages of pod development.     

Effect of nucleopolyhedrovirus infection of Spodoptera litura larvae on host discrimination by Microplitis pallidipes

The behaviour and oviposition of solitary endoparasitoid Microplitis pallidipes Szepligeti (Hymenoptera: Braconidae) were monitored to investigate the ability of the parasitoids to distinguish between nucleopolyhedrovirus (NPV)-infected and noninfected Spodoptera litura Fabricius (Lepidoptera: Noctuidae) larvae. The results indicated that the parasitoid searching time and the time until the first parasitoid attack on infected larvae were greater than those recorded on noninfected larvae; the number of infected larvae attacked by parasitoids, the percent of first attacks and parasitism rate in infected larvae were lower than those on noninfected larvae; and these differences were all significant 3 to 5 days postexposure of the larvae to a dose of 1.6 × 10⁸ occlusion bodies (OB)· ml^?1 and significant 4 and 5 days postexposure of the larvae to a dose of 1.6 × 10⁷ OB·ml^?1. The lowest dosage (1.6 × 10⁶ OB·ml^?1) had no significant effect on the above index values. In a field cage experiment, we found that the percentage of infected larvae parasitized by M. pallidipes gradually decreased as the time after NPV inoculation (1.6 × 10⁸ OB·ml^?1) increased, and that M. pallidipes significantly preferred to oviposit in healthy larvae from day 3 to day 5 after virus inoculation. Our research concluded that this parasitoid's ability to discriminate between healthy and infected hosts increased as virus concentration increased and as the time between exposure of hosts to virus and subsequent exposure to parasitoids increased.     

Partial purification and some properties of wineberry latent, a virus obtained from Rubus phoenicolasius

Wineberry latent virus (WLV) was obtained from a single symptomless plant of American wineberry (Rubus phoenicolasius) originally imported from the United States of America. On graft inoculation, WLV infected but induced no distinctive symptoms in several Rubus species including those used as indicators for known Rubus viruses. It was not seed-borne in wineberry. WLV was mechanically transmitted to several herbaceous species but induced local lesions in only a few; it was weakly systemic in some Chenopodium species. Infective C. quinoa sap lost infectivity after diluting to 10^-4, heating for 10 min at 70°C, and storage either for 8 days at 18°C or for 32 days at 4°C. Sap from infected plants contained flexuous filamentous particles c. 510°12 nm. WLV was partially purified by extracting infected C. quinoa leaves in 0·05 M tris-HCl buffer (pH ₇) containing 0·2% thio-glycerol and 10% (v/v) chloroform and concentrating virus by precipitation with 7% (w/v) polyethylene glycol (PEG, mol. wt 6000) and 0·1 NaCl. The virus was then pelleted through a 30% (w/v) sucrose pad containing 7% PEG+0·1 M NaCl and finally sedimented through a sucrose density-gradient. These preparations had A_260/280 ratios of 1·26, contained end to end aggregates of WLV particles and formed a partly polydispersed peak in the analytical ultracentrifuge. WLV did not react with antisera to four potex-viruses, or to apple chlorotic leaf spot or apple stem grooving viruses.     

Experiments on integration of chemical and biological control of aphids on brussels sprouts

Two field experiments were made to test whether natural enemies would take over control of brussels sprouts aphids at the time when protection from a selectively acting, soil-applied systemic insecticide, menazon, began to fail. The natural enemies, notably Syrphidae, proved ineffective against Brevicoryne brassicae L. despite advantages given by the insecticide and by close planting, which greatly increased the ratio of numbers of syrphid eggs and larvae to aphids. Thus, development of the aphid population was little altered when infested plants were kept free of most natural enemies by hand removal. I lb/acre (1–12 kg/ha) of menazon applied as spot treatments to the soil at planting-out time decreased the number of overwintering parasite mummies by 70 % but such ‘mortality’ was compensated for by decreased mortality from hyperparasites and other causes, so the numbers of adults of the primary parasite Diaretiella rapae (McIntosh) which emerged in the spring were similar to those from untreated plots. Soil cultivation in winter drastically decreased the numbers of emerging adult primary parasites, hyperparasites and syrphids. The menazon treatment designed for integrated control (1 lb/acre) seemed too unpredictable in action, e.g. less effective in dry than in damp conditions, to provide the hoped-for chemical control needed until natural enemies became abundant. Menazon at 4 lb/acre (4·48 kg/ha) protected the crop throughout the growing season and bioassays showed that menazon or its toxic derivatives continued to occur in leaves during cool periods in winter and also in the following April-May nearly 1 year after application. The same amount of menazon per unit area of crop was less effective on 1 ft 6 in (45·6 cm) spaced than on 3 ft (91·2 cm) spaced brussels sprouts plants.     

LIGHT,TEMPERATURE AND PHOTOPERIOD AS FACTORS CONTROLLING REPRODUCTION IN ULOTHRIX ZONATA (ULVOPHYCEAE)1

In many lakes in the northern United States and Canada the filamentous green alga Ulothrix zonata (Weber and Mohr) Kütz grows abundantly in early spring in shallow waters. Asexual reproduction occurs by formation of quadriflagellate zoospores which disrupt, the integrity of the cells upon release causing the filament to disintegrate. Study of the effects of 100 different combinations of irradiance, temperature and photoperiod revealed that zoospore formation is favored by high temperatures near 20°C, high light levels of 520 μE·m^?2·s^?1 and photoperiods of either short day (8:16 h light-dark) or long day cycles (16:8 h light-dark). Zoospore formation is minimal under conditions of low temperature (5°C), low irradiance (32.5 μE·m^?2·s^?1) and neutral day-lengths (12:12 h light-dark). These observations explain the decline in U. zonata biomass when water temperatures rise above 10° C. The combined effect of rising water temperatures and increasing daylengths causes progressively more filaments to switch from vegetable growth to zoospore production resulting in an increasing loss of biomass.     

Effects of Ozone Exposure on Resistance of Wheat Genotypes to Pyrenophora tritici-repentis

Three spring wheat genotypes, susceptible, moderately resistant or resistant to Pyrenophora tritici-repentis (tan spot fungus) were exposed to charcoal-filtered air and to approx. 80, 160, 240 (g m^?3 ozone for five consecutive days (7 h per day). Visible leaf injury on seedling plants (three-leaf stage) was only observed after fumigation with 160 or 240 (g m^?3 O₃. Amount of injury was four-fold and 10-fold on the susceptible genotype when compared to resistant or moderately resistant genotype at the two highest concentration of ozone, respectively. Genotypic differences to O₃ tolerance were detected at the seedling growth stage (three-leaf stage) and flowering stage but not at the stem elongation stage. A significant increase in tan spot lesion area was observed only on O₃ predisposed second top most leaves of the susceptible genotype at all the three levels of ozone. Predisposition did not enhance tan spot development in resistant and moderately resistant genotypes. In a test with 12 wheat genotypes, a highly significant positive correlation (r = 0· 986, p < 0· 0001) was observed between ozone sensitivity (percent leaf area damaged due to 240 (g m^?3 ozone exposure) and tan spot development (mm² lesion area) following inoculation with P. tritici-repentis. It indicates that wheat genotypes resistant to the tan spot fungus might be tolerant to ozone damage.     

Microscopic and biochemical evidence of differentially virulent field isolates of Diplocarpon rosae causing black spot disease of roses

Black spot disease caused by Diplocarpon rosae is one of the most widespread diseases of roses that are very difficult to control due to the generative reproduction and complex genetic constitution of roses and up to now the control of black spot still requires intensive use of systemic fungicides. Here we report for the first time evidence of differentially virulent field isolates of D. rosae. Using a combination of fungal structures, disease symptoms and host cells protein expression pattern analysis we here provide direct biochemical evidence that tropical field isolates of D. rosae are more virulent and caused disease symptoms earlier than their temperate counterparts. The tropical fungal field isolates strongly induced an excessive accumulation of ROS and repressed activity of pathogenesis-related proteins such as peroxidases, chitinase and phenylalanine ammonia lyase compared to their temperate counterparts. These findings bring insights into a hidden pathogenic characteristic of tropical D. rosae field isolates compared to their temperate counterparts and open a novel dimension of parameters to be considered when controlling black spot disease of roses by fungicides in tropical versus temperate regions. Interestingly, we found that treatment of rose leaves with ROS (H₂O₂) prior to fungal inoculation promoted plant defense response regardless of the isolate virulence based on protein expression pattern analysis, suggesting that ROS (H₂O₂) can be efficiently incorporated into black spot disease management.     

Bioassay method for assessing the virulence of Beauveria bassiana against tarnished plant bug, Lygus lineolaris (Hem., Miridae)

A standard bioassay method for assessing the pathogenicity of Beauveria bassiana (Balsamo) Vuillemin (GHA strain) against second instar tarnished plant bug, Lygus lineolaris (Palisot de Beauvois) (Hem., Miridae) was developed. Several types of inoculation methods, assay containers and incubation times were tested. Our goal was to minimize control mortality and maximize treatment mortality. Five inoculation methods (immersing broccoli florets or bean pods, spraying broccoli florets or bean pods, and immersing insects) and four types of plastic containers (114‐, 171‐, 228‐ and 455‐ml) were tested. Immersing insects directly in a fungal suspension was the most effective inoculation method, which resulted in a treatment mortality of 70–81.3% at a concentration of 1 × 10⁷ conidia/ml. The 114‐ml plastic container was the most suitable assay container when 10 tarnished plant bug nymphs were treated together, resulting in a control mortality of only 6% 12 days after treatment. Within the first 6 days after treatment, 71.1% of the insects were killed, compared with a total mortality of 81.3% after 12 days. Nymphs infected with the test fungus changed colour from green to black. Mycelial outgrowth and sporulation on the cadavers demonstrated that most nymphs died of fungal infection. A total of 61.1 and 80.5% of the cadavers showed signs of mycelial outgrowth 9 days after death among those that were surface sterilized and those that were not, respectively.     

A new experimental model of acute osteomyelitis due to methicillin-resistant Staphylococcus aureus in rabbit

Aims: To assess the impact of antibiotic therapy on severe osseous infections, animal models of chronic bacterial infections have been developed; however, these models suffer from many experimental limitations. The aim of this work was to develop a new model system in which high levels of bacteria are obtained within femoral bone marrow and bone tissue, and such infections are maintained for at least 14 days. Methods and Results: Experimental osteomyelitis was induced in 25 New Zealand white rabbits. A 10⁹ CFU ml^?1 suspension of methicillin‐resistant Staphylococcus aureus was injected into the knee after bone trepanation. On day 3, surgical debridement was performed to mimic a surgical procedure. Animals were euthanized 1, 2, 3, 9 and 14 days post‐inoculation to determine the bacterial counts in marrow and bone, and to evaluate the stability of the infection. Inoculated lesions also were assessed for changes in histological parameters on days 3 and 7 post‐inoculation. At days 1, 2, 3, 9 and 14 post‐inoculation, we observed 6·50 ± 0·64, 7·30 ± 0·49, 7·82 ± 0·19, 8·00 ± 1·48 and 8·99 ± 0·20 log10 CFU g^?1 in bone marrow and 8·40 ± 0·68, 7·65 ± 0·27, 7·58 ± 0·30, 8·88 ± 0·52 and 8·28 ± 0·39 log10 CFU g^?1 in bone tissue, respectively. No statistical differences in bacterial count were found between bone marrow and bone tissue at any time point. Conclusion: This new model of acute osteomyelitis was validated by histological and microbiological changes in the absence of sclerosing agents, and these changes remained stable for 14 days. Significance and Impact of the Study: These results describe a new experimental model of acute osteomyelitis and demonstrate its usefulness in assessing the activity of antibacterial agents in vivo soon after bone infection.