The ultrastructure of germinating conidia of Penicillium digitatum inhibited by sec-butylamine

Electron micrographs of germinating conidia of P. digitatum showed osmiophilic inclusions scattered in the ground plasm or located within membrane-bound vacuoles where they were associated with various membranes and “myelin-like” structures. Cytochemical studies suggested that the osmiophilic inclusions within the vacuoles contained phospholipids and proteins. These phospholipoprotein inclusions may provide precursors needed for the assembly of membrane-bound organelles formed during germination. In conidia germinated in the presence of fungistatic concentrations of sec-butylamine (SBA) the vacuoles enlarged and the phospholipoprotein inclusions decreased in size and appeared to disintegrate and give rise to membrane-bound vesicles. SBA may deprive the conidium of materials normally utilized for membrane formation by the developing organelles.     

Influence of forest clearing methods on nutrient uptake of some tropical food crops

The adverse effects of forest clearing on crop yield have often been directly ascribed to decline in soil fertility. However, the effect of various forest clearing methods on crop nutrient uptake as reflected by the content in the leaf tissues has been the subject of few scientific investigations. A study to determine the effect of forest clearing on nutrient content of some arable crops was, therefore, carried out in the tropical rainforest of south-western Nigeria. Three forest clearing methods—mechanical, semi-mechanical and manual clearing—were used, while maize, cowpea, and cassava were the test crops. Nutrient content in the tissues of these crops was assessed by sampling maize ear leaves at silking, youngest mature leaves (YML) of cowpea at flowering and YML of 3-month-old cassava for analysis. Correlation analysis was carried out to establish the relationship between nutrient content and crop yield. The method of forest clearing had no significant effect on the nutrient content of maize and cowpea during the first cropping cycle. However, nutrient content of maize in the manually cleared plots was higher than those in the semi- and mechanically cleared plots for all elements analyzed except Mg. Nitrogen content of cowpea was much higher than that of maize. Except for P and K, the nutrient content of cassava was not markedly different among the forest clearing methods. Calcium and P content in the leaves of maize and cassava varied inversely with their levels in the soil, implying that crop uptake could have accounted for the differences in their availability within the soil. Nitrogen, P and K had significant positive correlations with maize yield (r = 0.77, 0.89, and 0.71). Potassium content also had significant positive correlation with cassava root yield (r = 0.69). This is not surprising because compared with other crops, cassava requires large amounts of K for starch formation and root development. It is concluded that differences observed in crop yields under different forest clearing methods are a function of nutrient uptake and availability within the soil. However, the uptake of some nutrients does not seem to be determined by the level of soil availability alone. This study was supported with a research grant from the International Foundation for Science (IFS), Sweden.     

An inoculation experiment of Japanese <Emphasis Type="Italic">Bursaphelenchus</Emphasis> nematodes on Japanese black and red pine, <Emphasis Type="Italic">Pinus thunbergii</Emphasis> and <Emphasis Type="Italic">P. densiflora</Emphasis>

The pinewood nematode (PWN) Bursaphelenchus xylophilus is an invasive pathogen that was introduced from North America to Asian countries and Portugal and is devastating native pine forests. Some native European and Asian Bursaphelenchus nematodes also have weak to moderate pathogenicity to native pine species. To evaluate the potential risk of native Bursaphelenchus species, we inoculated ten Japanese Bursaphelenchus species into native pine species (the dominant forest species) in Japan, and evaluated their pathogenicity using mortality and tracheal tissue damage as indices. Inoculation was conducted on August 3, 2007, and the symptoms were observed every 2 weeks until February 1, 2008. None of the inoculated trees, excluding the pathogenic PWN inoculated control, showed external disease symptoms; however, four species [a less pathogenic PWN isolate, B. luxuriosae, Bursaphelenchus sp. NK215 (undescribed), and NK224 (undescribed)] caused tracheal tissue damage in inoculated seedlings and showed weak pathogenicity. Therefore, we conclude that there are some potentially pathogenic native species of nematodes distributed in Japan. Interestingly, two of these weakly pathogenic species, B. luxuriosae and NK215, are not associated with Pinaceae trees, suggesting that nematode pathogenicity may be a pre-adaptive character. More experimental studies under different conditions are necessary to accurately evaluate the potential risk of these pathogens.     

Possible Role of ZPAC,Zygote-specific Proteasome Assembly Chaperone,During Spermatogenesis in the Mouse

In the mammalian testis, the ubiquitin-proteasome system plays important roles in the process that promotes the formation of mature sperm. We recently identified zygote-specific proteasome assembly phaperone (ZPAC), which is specifically expressed in the mouse gonads and zygote. ZPAC mediates a unique proteasome assembly pathway in the zygote, but the expression profile and function of ZPAC in the testis is not fully understood. In this study, we investigated the possible role of ZPAC during mouse spermatogenesis. First, we analyzed the expression of ZPAC and 20S proteasome subunit α4/PSMA7 in the adult mouse testis. ZPAC and α4 were expressed in spermatogonia, spermatocytes, and round spermatids. In elongating spermatids, ZPAC was expressed until step 10, whereas expression of α4 persisted until step 12. We then examined the expression profile of ZPAC and α4 in a mouse model of experimental unilateral cryptorchidism. Consistent with appearance of morphologically impaired germ cells following cryptorchidism, the ZPAC protein level was significantly decreased at 4 days post induction of experimental cryptorchidism (D4) compared with the intact testis, although the amount of α4 protein persisted at least until D10. Moreover, intense ZPAC staining was co-localized with staining of annexin V, an early indicator of apoptosis in mammalian cells, in germ cells of cryptorchid testis, but ZPAC was also expressed in germ cells showing no detectable expression of annexin V. These results suggest that ZPAC plays a role during spermatogenesis and raises the possibility that 20S proteasome mediated by ZPAC may be involved in the regulation of germ cell survival during spermatogenesis.     

Measurement of polynuclear aromatic hydrocarbons in canine lung after alkaline decomposition

An alkaline decomposition method employing a KOH/alcohol solution was studied, and polynuclear aromatic hydrocarbons (PAHs) contained in particles remaining in canine lung were measured. As a result, BaA, BkF, BaP, and BghiP were found. By this method, PAHs extracted from the lungs of 32 dogs were 13.0-166.0 ng (mean, 63.0 ng) for BaA, 6.6-90.2 ng (mean, 27.4 ng) for BkF, 9.8-167.4 ng (mean 47.2 ng) for BaP, and 10.8-206.0 ng (mean, 61.8 ng) for BghiP. The results showed no correlation between the age and the concentration of PAHs in the lung, but some correlation was found between the age and the lung weight (p<0.01). There were significant correlations among the concentrations of the compounds in the lung (p<0.01). These results suggest that dogs, like humans, are affected by automobile exhaust and other common generation sources of such substances.     

Production of lipid mediators in mastitic milk of cow

Bovine mastitis is one of the most prevalent and costly diseases in the dairy industry. Lipid mediators are signaling molecules which coordinately and intricately modulate inflammation. They are produced from polyunsaturated fatty acids (PUFAs) in the cellular membrane via several enzymes including cyclooxygenase (COX) and lipoxygenase (LOX). In the present study, we performed comprehensive analysis of lipid production in milk obtained from clinical or subclinical mastitic cows using liquid chromatography/mass spectrometry. We detected 26, 24, and 40 kinds of lipid constantly in healthy, subclinical, and clinical mastitic milk, respectively. In clinical mastitic milk, the amount of a major n‐6 PUFA, arachidonic acid (AA), tended to increase, whereas amounts of major n‐3 PUFAs, eicosapentaenoic acid and docosahexaenoic acid, tended to decrease. The amounts of several AA‐derived lipids including COX‐catalyzed prostaglandin (PG) D₂ and PGE₂, and LOX‐catalyzed leukotriene (LT) B₄ were increased in clinical mastitic milk. Although subclinical mastitic milk represented similar trend of lipid production to healthy milk, amounts of several lipids such as LTD₄, 14,15‐dihydroxyeicosatrienoic acid, and 14‐epoxyeicosatrienoic acid changed. These findings would be helpful for better understanding of mastitis pathology and give us some insights to develop a new diagnostic and therapeutic strategy.     

Cloning of the gene coding for Staphylococcus intermedius exfoliative toxin and its expression in Escherichia coli

An exfoliative toxin (SIET)-producing strain (D-52) of Staphylococcus intermedius derived from canine pyoderma did not possess large plasmids. Therefore, the gene coding for SIET was considered to be located on the chromosomal DNA. The SIET gene was cloned from the chromosomal DNA of S. intermedius and was expressed in Escherichia coli. The nucleotide sequence of the SIET gene consists of a coding region of 990 bp specifying a polypeptide of 330 amino acid residues, which included a putative 42-residue signal sequence.     

Molecular cloning and expression of bottlenose dolphin (Tursiops truncatus) interleukin-8

The bottlenose dolphin interleukin (IL)-8 cDNA was molecularly cloned. The dolphin IL-8 has an open reading frame of 303-bp encoding 101 amino acids. The homology of the amino acid sequence with that of other species was: sheep, 89.1%; cattle, 88.1%; pig, 85.1%; dog, 85.1%; horse, 79.2%; human, 74.5%; and macaque, 72.3%. The amino acid sequence suggested that dolphin IL-8 was a CXC chemokine. The recombinant dolphin IL-8 protein was recognized with anti-ovine IL-8 monoclonal antibody.