The pathogenic potential of endophytic Botryosphaeriaceous fungi on Terminalia species in Cameroon

In Cameroon, native Terminalia spp. represent an important component of the forestry industry, but limited information is available regarding the fungal pathogens that affect them. The Botryosphaeriaceae are endophytic fungi and latent pathogens that can result in wood stain, cankers, die‐back and death of trees, particularly when trees are under stress. The aim of this study was, therefore, to identify and characterize the Botryosphaeriaceae occurring as endophytes of Terminalia spp. in Cameroon, as part of a larger project to identify potential pathogens of these trees in the country. Samples were collected from three Terminalia spp. in the Central, Southern and Eastern Regions and the resultant Botryosphaeriaceae were identified using morphology and DNA sequence comparisons for the ITS and tef 1‐α gene regions. Furthermore, inoculation trials were conducted to consider the relative pathogenicity of the isolates collected. The majority of isolates (88%) represented species of Lasiodiplodia, including L. pseudotheobromae, L. theobromae and L. parva. The remaining isolates were identified as Endomelanconiopsis endophytica. Pathogenicity trials on young T. mantaly and T. catappa trees revealed that L. pseudotheobromae was the most pathogenic species followed by L. theobromae.     

Biochemical composition and kinetics of C and N mineralization of animal wastes: a typological approach

Forty-seven different animal wastes were characterized using chemical and organic matter fractionation methods (water extraction and Van Soest method) and 224-day incubation studies to assess their decomposition in soil. Simple correlation and multiple factor analysis were performed to establish relationships between the composition of these wastes and C and N mineralization. Carbon and N contents ranged from 101 to 469 mg C kg⁻¹ dry matter (d.m.) and from 4 to 39 mg N kg⁻¹ d.m. Soluble C and N represented less than 9% of organic C and 1.5% of total N at 20°C, respectively. The C fractions soluble at 100°C or in neutral detergent were larger and represented 14 and 32% of the organic C, respectively. The hemicellulose-like (HEM) and cellulose-like (CEL) fractions contained about 16.5 and 6% of the organic N, respectively. The C distribution in the lignin-like (LIG) and CEL fractions was comparable, but the former contained more N. Carbon mineralization varied from 5 to 62% of the organic C added during the 224-day incubation; 70% of the wastes induced net N mineralization at the end of incubation (from 3 to 51% of organic N). Other wastes induced net soil inorganic N immobilization, from −1 to −31% of the organic N added. Most highly significant correlations were established between the C mineralization and the C present in the water-soluble fraction at 20°C, and the HEM and LIG fractions. Relationships between N mineralization and biochemical characteristics were weak, except with the soluble Van Soest fraction, and highly significant correlations were observed between N mineralization rates calculated at 224 days of incubation and the organic N content or C/N ratio of wastes. Finally, an objective hierarchical classification based on composition criteria and C and N mineralization led to the definition of six different classes of wastes. It permitted differentiation between four composted wastes and intrinsically different wastes (i.e., cattle manures, pig manures, and poultry manures) which could not be objectively regrouped. It also placed some very different types of waste (solid phase from pig slurry separation, pig manures, and composted pig mixtures) in the same class.     

Abiotic degradation of lignified cell walls by carbonate and copper salt

This study reports on the destructuration of Wheat straw and Spruce wood cell walls after maceration in potassium carbonate or sodium hydroxide at pH = 10 in the presence of copper acetate. The alkaline treatments had a predominant impact on the wheat straw cell wall components over copper acetate. Either K-carbonate or Na-hydroxide extracted from wheat straw a particular lignin fraction rich in condensed C-C linkages, leading to the unmasking of new ether-linked sub-structures in the cell wall. This unmasking was increased in the presence of copper salt but only in the nonextracted Wheat straw sample incubated in carbonate and not in the corresponding extractive-free sample. This difference was related to the leaching of compounds from the nonextracted cell wall, which could sustain oxidative activity of copper by hindering its precipitation into inactive hydroxide and/or carbonate species. In Spruce wood samples, copper salt was the principal factor impacting on the lignin structure over alkali alone. Its effect was, however, only detected at the level of C-C linked dimers. These results confirmed that unmasking of lignin sub-structures also occurred in Spruce wood, but probably through mechanisms different from that evidenced in Wheat straw.     

Possible role of plant phenolics in the production of trichothecenes by Fusarium graminearum strains on different fractions of maize kernels

Four trichothecene-producing strains of Fusarium graminearum were grown on three maize grain fractions, whole grain, degermed grain, and the germ, to determine the effect of natural substrates on mycotoxin production. Monitoring the ergosterol content after 25 days of incubation indicated that fungal growth on all grain fractions was comparable. Trichothecene (TCT) production was highest on degermed grain, less on whole grain, and very low or nondetectable on the germ; similar results were found with four different strains. It was concluded that inhibitor(s) of TCT biosynthesis were present in maize germ. The presence of phenolic compounds was investigated in the different fractions. The hydroxamate 4-acetylbenzoxazolin-2-one (4-ABOA), a known inhibitor of mycotoxin production, was found in the degermed and whole grain fractions but not in the germ. Therefore, the TCT inhibition observed on the maize germ fraction used in our study is clearly not linked to 4-ABOA. Other soluble phenolic compounds were found at a much higher concentration in the germ than in the two other fractions. The inhibition property of the soluble ester-bound extracts was tested in liquid culture. A possible role for these compounds is discussed.     

The origin of the A genome donor of wheats (Triticum: Poaceae) – a perspective based on the sequence variation of the 5S DNA gene units

In a prior study on the haplomes of wheat using the 5S rRNA gene we assigned the long A1 and short A1 unit classes to the A haplome in the diploid T. monococcum. The short A1 unit class is absent in the tetraploids T. turgidum and T. timopheevii and in the hexaploid T. aestivum, although present in the hexaploid T. zhukovskyi. Both T. turgidum and T. aestivum contained a different 5S DNA unit class labeled the short A2.The purpose of this paper was to study the short A2 units in the two diploid species to shed light on the theory that the A haplome donor of T. turgidum and T. aestivum was T. urartu. Fifty eight clones were obtained from 12 accessions, sequenced and analyzed. As expected T. baeoticum, which is often classified as a subspecies of T. monococcum, contained the long A1 and the short A1 5S DNA units. Unexpectedly, T. urartu had the long A1 and the short G1 unit classes instead and other units not found so far in Triticum. These findings support the hypothesis that the donor of the A genome in T. zhukovskyi was T. monococcum, as identified by the short A1 units. However, the short A1 units are absent in T. timopheevii, also a carrier of the A genome. The short G1 units found in T. urartu also identify it as a possible donor of the G genome to T. timopheevii. The short G1 units were also found in T. aestivum in our prior study. The long G1 unit class was not found in T. urartu but reported from T. timopheevii and T. zhukovskyi. The implications of these and related findings on the evolution of wheats are discussed.     

Multiloculated solitary (unicameral) bone cyst in a young dog

A 20‐month‐old female spayed Staffordshire Terrier (22.3 kg) presented to the Orthopedic Surgery Service at North Carolina State University Veterinary Teaching Hospital for evaluation of a 6‐week history of toe‐touching to nonweight‐bearing lameness in the right hind limb. Radiographs of the right stifle revealed a multiloculated lytic lesion of the distal femur, with a large open lytic zone centrally, numerous osseous septations peripherally, and focal areas of cortical thinning and loss. An aspirate of the right distal femoral lesion yielded mildly cloudy serosanguineous fluid. Cytologic examination of the fluid revealed a pleomorphic population of discrete cells that exhibited marked anisocytosis and anisokaryosis and a variable nuclear‐to‐cytoplasmic (N:C) ratio, which were interpreted as probable neoplastic cells, with few macrophages, and evidence of hemorrhage. Given the clinical signs of pain, lesion size, and concern for malignant neoplasia, amputation of the right hind limb was performed. Histologically, the lesion had undulating walls 1‐3 mm thick with a continuous outer layer of dense fibrous tissue and an inner layer composed of reactive cancellous bone with no cortical compacta remaining. Remnants of thin fibrous or fibro‐osseous septa projected from the bony wall into the cyst lumen. The final histologic diagnosis was a benign multiloculated solitary (unicameral) bone cyst of the distal right femur. Based on the histopathologic findings, it was speculated that the cells identified on cytology were a mixture of developing osteoclasts, osteoblasts, endothelial, and stromal cells. This is the first report describing the cytologic examination of a solitary bone cyst in veterinary medicine.     

Immunoproteomic analyses of outer membrane antigens of Actinobacillus pleuropneumoniae grown under iron-restricted conditions

Actinobacillus pleuropneumoniae, a bacterial pathogen of swine and agent of porcine pneumonia, causes a highly infectious disease of economic importance in the pig industry. Commercial vaccines for A. pleuropneumoniae include whole-cell bacterins and second generation subunit vaccines but they only confer partial protective immunity. Our search for new vaccine candidates identified antigens that are expressed during conditions that mimic infection; the outer membrane (OM) proteome of A. pleuropneumoniae serotype 5b was examined under iron restriction. Quantitative profiling by 2D-DiGE technology revealed that iron restriction induced expression of previously described transferrin binding proteins (TbpA, TbpB) plus four lipoproteins including spermidine/putrescine binding periplasmic protein 1 precursor (PotD2). Immunoproteomic analyses with antisera from na?ve animals and from infected pigs were able to differentiate antigens within the OM proteome that were specifically recognized during A. pleuropneumoniae infection. Immunoblots of iron-restricted profiles detected PotD2, heme-binding protein A (HbpA), and capsule polysaccharide export protein (CpxD) as well as surface antigens TbpA, TbpB, and OmlA. These data identify OM proteins that demonstrate immunogenicity and upregulation under conditions mimicking infection, providing emphasis on lipoproteins as an important class of antigens to exploit for vaccine development for A. pleuropneumoniae.     

Adaptation de la teneur en enzymes digestives de l'hepatopancreas de Palaemon serratus (Crustacea,Decapoda), a la composition d'aliments experimentaux

The effects of the carbohydrate and protein composition of compound diets on growth rate and on the amylase and protease content of the hepatopancreas have been studied in Palaemon serratus.The best growth rate for animals weighing between 80 and 380 mg is obtained with a diet containing 3.5% carbohydrate and 64% protein. Diets which are very rich in carbohydrate and poor in protein result in poor growth and high mortality; the same is true for foods containing a high concentration of only one protein.Variations in the wet weight of the hepatopanereas and in the total digestive enzymatic activities are linked to variations in the growth rate of the shrimp.Maximal specific amylase activity is reached when assimilable carbohydrate amounts to 2.8%. Maximal specific protease activity is reached when the food protein level reaches 45%. When the amounts of carbohydrate and protein are higher or lower, the values of specific enzymatic proteins diminish.     

Characterization of Botryosphaeriaceae from plantation‐grown Eucalyptus species in South China

The Botryosphaeriaceae is a species‐rich family that includes pathogens of a wide variety of trees, including Eucalyptus species. Symptoms typical of infection by the Botryosphaeriaceae have recently been observed in Eucalyptus plantations in South China. The aim of this study was to identify the Botryosphaeriaceae associated with these symptoms. Isolates were collected from branch cankers and senescent twigs of different Eucalyptus spp. All isolates resembling Botryosphaeriaceae were separated into groups based on conidial morphology. Initial identifications were made using PCR‐RFLP fingerprinting, by digesting the ITS region of the rDNA operon with the restriction enzymes CfoI and KspI. Furthermore, to distinguish isolates in the Neofusicoccum parvum/N. ribis complex, a locus (BotF15) previously shown to define these species, was amplified and restricted with CfoI. Selected isolates were then identified using comparisons of DNA sequence data for the ITS rDNA and translation elongation factor 1‐alpha (TEF‐1α) gene regions. Based on anamorph morphology and DNA sequence comparisons, five species were identified: Lasiodiplodia pseudotheobromae, L. theobromae, Neofusicoccum parvum, N. ribis sensu lato and one undescribed taxon, for which the name Fusicoccum fabicercianum sp. nov. is provided. Isolates of all species gave rise to lesions on the stems of an E. grandis clone in a glasshouse inoculation trial and on the stems of five Eucalyptus genotypes inoculated in the field, where L. pseudotheobromae and L. theobromae were most pathogenic. The five Eucalyptus genotypes differed in their susceptibility to the Botryosphaeriaceae species suggesting that breeding and selection offers opportunity for disease avoidance in the future.