Molecular and serological prevalence of Babesia bovis and Babesia bigemina in water buffaloes in the northeast region of Thailand

Bovine babesiosis is a tick-transmitted hemoprotozoan disease that is mainly caused by Babesia bovis and Babesia bigemina and is characterized by significant morbidity and mortality worldwide. The disease is widespread in the northeastern region of Thailand, where an increasingly large part of the livestock is composed of water buffaloes. The present study was therefore conducted to investigate the epidemiological distribution of B. bovis and B. bigemina in water buffaloes in the northeastern region of Thailand. A total of 305 buffalo blood samples were randomly collected from five provinces and simultaneously analyzed by the nested PCR (nPCR) assay, ELISA, and IFAT techniques. The overall prevalence of B. bovis and B. bigemina was 11.2% and 3.6% by nPCR, 14.7% and 5.9% by ELISA, and 16.8% and 5.6% by IFAT, respectively. The high concordance between the molecular and the serological detection tests revealed the specificity and sensitivity of the diagnostic assays used for the detection of infection as well as the endemic stability status of the parasites in the surveyed areas. Statistically significant differences in the prevalence of the two infections were observed on the basis of age and location but not gender. Our data provide valuable information regarding the epidemiology of B. bovis and B. bigemina infection in water buffaloes in the northeastern region of Thailand which will likely be very beneficial for management and control programs of this disease.     

Genes expressed in Japanese flounder <Emphasis Type="Italic">Paralichthys olivaceus</Emphasis> spleen: analysis of genes involved in immune function

ABSTRACT:   Expressed sequence tag (EST) analysis is an efficient tool for gene discovery and for profiling gene expression. A cDNA library developed from messenger RNA of the Japanese flounder, Paralichthys olivaceus spleen was constructed by directional cloning, in order to isolate functional genes involved in immunity in fish. A total of 1010 ESTs from the library were sequenced and compared with sequences in the GenBank database. Of the 1010 ESTs, 618 ESTs (61%) were identified as being homologous with known genes from many organisms by BLAST searches, whereas 392 (39%) appeared to be unknown and are likely to represent newly described genes. Of the identified genes, 105 (17%) encoded proteins associated with cell/organism defence and homeostasis. Of these 105 genes, 21 were identified for the first time in Japanese flounder. These included macrophage inflammatory protein (MIP)-3 α, granulin-A, novel immune-type receptor (NITR), interferon regulatory factor (IRF)-10, presenilin-like protein-2, and antizyme inhibitor. A comparison of ESTs derived from spleen, kidney, liver and leukocytes suggested that expression of immune-related genes in different tissues, organs, or cells are different.     

New type of heat shock protein 70 homologue gene abounds in the genomic sequence of kuruma shrimp Marsupenaeus japonicus

Heat shock proteins (HSPs) are a group of highly conserved molecular chaperones. Shrimp HSPs have recently been a topic of increasing interest because of their roles in shrimp immunity and homeostasis. In penaeid shrimp, HSP70s and the cognate forms, heat shock cognate (HSC) 70s, have been reported, but their responses towards various stimulations are different. We found a novel type HSP70 (MjHSP70-2) from the hyperexpansion of the large segmental duplication that is present in kuruma shrimp Marsupenaeus japonicus, which shows about 60 % identity with reported shrimp HSP70s. In a phylogenetic tree, MjHSP70-2 formed a sister clade with eukaryote HSP70 family while MjHSP70 was located close to the shrimp HSP70 and HSC70 group. MjHSP70-2 gene expression was not significantly increased by heat shock or pathogen challenge by Vibrio penaeicida, but it was significantly increased by infection with white spot syndrome virus. In contrast, MjHSP70 gene expression was increased by heat shock but decreased by infection with V. penaeicida. The kuruma shrimp genome was found to have 400-fold more copies of the MjHSP70-2 gene than the putative single-copy gene transglutaminase. In conclusion, our results reveal the presence of a novel type HSP70 gene, HSP70-2, from kuruma shrimp. There are multiple forms of HSP70 in crustaceans, and these HSP70s behave differently under various stressors.     

Heat-epimerized tea catechins rich in gallocatechin gallate and catechin gallate are more effective to inhibit cholesterol absorption than tea catechins rich in epigallocatechin gallate and epicatechin gallate

It has been known that tea catechins, (-)-epicatechin (1), (-)-epigallocatechin (2), (-)-epicatechin gallate (3), and (-)-epigallocatechin gallate (4) are epimerized to(-)-catechin (5), (-)-gallocatechin (6), (-)-catechin gallate (7), and (-)-gallocatechin gallate (8), respectively, during retort pasteurization. We previously reported that tea catechins, mainly composed of 3 and 4, effectively inhibit cholesterol absorption in rats. In this study, the effect of heat-epimerized catechins on cholesterol absorption was compared with tea catechins. Both tea catechins and heat-epimerized catechins lowered lymphatic recovery of cholesterol in rats cannulated in the thoracic duct and epimerized catechins were more effective than tea catechins. The effect of purified catechins on micellar solubility of cholesterol was examined in an in vitro study. The addition of gallate esters of catechins reduced micellar solubility of cholesterol by precipitating cholesterol from bile salt micelles. Compounds 7 and 8 were more effective to precipitate cholesterol than 3 and 4, respectively. These observations strongly suggest that heat-epimerized catechins may be more hypocholesterolemic than tea catechins.     

Fungal-feeding habits of six nematode isolates in the genus Filenchus

The family Tylenchidae is a large group of soil nematodes but their feeding habits are not fully known. We studied the fungal-feeding abilities of nematodes in the genus Filenchus. We measured population growth rates (PGRs) of six nematode isolates, representing three Filenchus species, when feeding on seven fungal species on two types of culture media. On Potato Dextrose Agar (PDA) Filenchus misellus, Filenchus discrepans and an unidentified Filenchus sp. generally showed moderate to large PGRs on saprophytic fungi (Rhizoctonia solani, Chaetomium globosum, Coprinus cinereus, Flammulina velutipes) and low PGRs on plant-pathogenic fungi (Fusarium oxysporum, Pythium ultimum). In soil medium amended with chopped soybean plant material or wheat bran, the status of most of the fungi as food for the nematodes was similar to that on PDA, although PGRs tended to be lower in the soil medium. However, C. globosum, a good food on PDA, only supported low PGR in soil for each of the three nematodes. The PGRs of F. misellus on C. globosum in soil were still low even when types and amounts of organic matter amendments were varied. A nematophagous fungus, Pleurotus ostreatus (oyster mushroom), was determined to be a food for Filenchus on PDA or in soil, based on PGR measurements corrected for extraction efficiency. To determine whether fungal species and culture media affected nematode extraction efficiencies and, consequently, the apparent PGRs, we compared efficiencies between R. solani, C. globosum and C. cinereus, and between PDA and soil. The relatively low extraction efficiencies across fungal species in soil seemed responsible for the lower nematode PGRs in soil than on PDA. On PDA generally, fungal species did not affect the assessment. In soil, effects of fungal species on extraction were significant, but not consistent, across nematode species. Nevertheless, the extraction efficiency differences in soil were considered not to affect assessment of the three fungi as food for the nematodes. The confirmation that three Filenchus species reproduce by feeding on fungi in soil suggests that fungal-feeding is not an unusual habit in the field, in this genus. We believe that in community studies, nematodes in the genus Filenchus should be considered fungal feeders or root and fungal feeders, rather than only plant feeders. Our confirmation of fungal-feeding habits in the genus Filenchus supports the hypotheses that plant-feeding nematodes evolved from those feeding on fungi.     

Retention of Rice dwarf virus by Descendants of Pairs of Viruliferous Vector Insects After Rearing for 6 Years

ABSTRACT Rice dwarf virus (RDV) is characterized by its unusual ability to multiply in both plants and leafhopper vector insects and by its transovarial mode of transmission. Colonies of Nephotettix cincticeps, derived originally from pairs of leafhoppers infected with an ordinary strain of RDV, were maintained for 6 years in the laboratory and were found, at the end of this time, still to harbor RDV. Moreover, the isolate of RDV, designated RDV-I, obtained from these colonies retained the ability to infect rice plants. When we raised leafhoppers separately from eggs that had been placed individually on pieces of water-soaked filter paper and reared them in the presence of healthy rice seedlings, we found that all of these leafhoppers harbored RDV. This observation suggested that RDV-I had been maintained in the leafhoppers by transovarial transmission. Two further observations, namely, the low rate of acquisition of RDV by virus-free insect nymphs on symptomless plants on which viruliferous insects had been reared, and the fact that only 2 to 5% of plants had symptoms when rice seedlings were inoculated via RDV-I-viruliferous insects, confirmed that the maintenance of RDV-I by any other mode of transmission through plants and insects was unlikely. This efficient and long-term maintenance of RDV in a population of viruliferous insects might explain the prolonged duration of rice dwarf disease in the field, once there has been a serious outbreak.     

Interspecific variation of photosynthesis and leaf characteristics in canopy trees of five species of Dipterocarpaceae in a tropical rain forest

Photosynthetic rate, nitrogen concentration and morphological properties of canopy leaves were studied in 18 trees, comprising five dipterocarp species, in a tropical rain forest in Sarawak, Malaysia. Photosynthetic rate at light saturation (Pmax) differed significantly across species, varying from 7 to 18 micro mol m(-2) s(-1). Leaf nitrogen concentration and morphological properties, such as leaf blade and palisade layer thickness, leaf mass per area (LMA) and surface area of mesophyll cells per unit leaf area (Ames/A), also varied significantly across species. Among the relationships with leaf characteristics, Pmax had the strongest correlation with leaf mesophyll parameters, such as palisade cell layer thickness (r2 = 0.76, P < 0.001) and Ames/A (r2 = 0.73, P < 0.001). Leaf nitrogen concentration and Pmax per unit area also had a significant but weaker correlation (r2 = 0.46, P < 0.01), whereas Pmax had no correlation, or only weakly significant correlations, with leaf blade thickness and LMA. Shorea beccariana Burck, which had the highest P(max) of the species studied, also had the thickest palisade layer, with up to five or more layers. We conclude that interspecific variation in photosynthetic capacity in tropical rain forest canopies is influenced more by leaf mesophyll structure than by leaf thickness, LMA or leaf nitrogen concentration.     

Runoff nitrogen from a large sized paddy field during a crop period

Nutrient load management is an important environmental issue because nutrient loads from farmlands degrade surface waters as a result of anthropogenic eutrophication. Nitrogen load from a large sized paddy field during the crop period was examined from the results of field measurements carried out in 2004. The 1.5 ha paddy field was located east of Biwa Lake. Irrigation water volume and ponded water depth were continuously observed. Field measurements were carried out at least once a week to analyze total nitrogen (TN) concentration in the irrigation water and ponded water. Daily inflow and outflow of nitrogen was obtained by multiplication of the nitrogen concentration and transported water volume, consisting of irrigation, precipitation, evapotranspiration, percolation and surface discharge. Water outflow volume was calculated by a tank model that consisted of three small tanks connected to represent ponded water depth differences in the large paddy field. The calculated nitrogen load was 18.8 kg ha⁻¹, with 7.2 kg ha⁻¹ from surface drainage and 11.6 kg ha⁻¹ from percolation loss. The runoff nitrogen value of 18.8 kg ha⁻¹ was within the range of the reported values investigated in a medium-sized paddy field. The observed value was close to the value for a low percolation flux paddy field where less irrigation water has been applied. These results suggest that less irrigation water keep runoff nitrogen low. This also indicates that irrigation water management can reduce nitrogen load from large sized paddy fields.