Antiparasitic activity of the medicinal plant Dillenia suffruticosa against the marine leech Zeylanicobdella arugamensis (Hirudinea) and its phytochemical composition

Zeylanicobdella arugamensis (Annelida: Hirudinea), a marine parasitic leech, is currently affecting different species of cultured groupers, hybrid groupers, snappers and sea bass in Malaysia. Dillenia suffruticosa (Dilleniaceae), a medicinal plant found in Sabah, has been selected in our experiment to kill the leeches as a natural control method. The leech‐infested hybrid groupers were collected from aquaculture facilities, and the isolated leeches were challenged against methanol extract of D. suffruticosa leaves. The experiment was carried out using various concentrations of the extracts such as 25, 50 and 100 mg/ml. The methanol extract showed significant antiparasitic activity against Z. arugamensis with 100% mortality at a concentration of 100 mg/ml in 14.39 ± 3.75 min., followed by 50 and 25 mg/ml in 32.97 ± 9.29 and 41.77 ± 5.40 min., respectively. The phytochemical composition of the extract was determined using GC‐MS analysis to understand the nature of the principal compounds responsible for its antiparasitic properties. The leaves of D. suffruticosa demonstrated the presence of different bioactive compounds of various natures with varying percentages. Thus, it could be revealed that the methanol extract of D. suffruticosa mainly contains vital phytochemical compounds and showed an effective antiparasitic activity against the harmful leeches of hybrid groupers.     

Distribution and habitat characteristics of three sympatric cetacean species in the coastal waters of Matang,Perak, Peninsular Malaysia

1. Coastal cetaceans in Southeast Asia are poorly studied and are particularly vulnerable to anthropogenic threats, especially in intensive fishing grounds.
2. To investigate the distribution and habitat characteristics of cetaceans in the productive coastal waters of Matang, Perak, Malaysia, boat‐based line transect surveys were conducted between 2013 and 2016.
3. The Irrawaddy dolphin (Orcaella brevirostris) was most frequently encountered at 3.87 sightings per 100 km, followed by the Indo‐Pacific finless porpoise (Neophocaena phocaenoides) at 1.72 sightings per 100 km, and the Indo‐Pacific humpback dolphin (Sousa chinensis) at 0.66 sightings per 100 km.
4. The mean group size was largest for humpback dolphins (8.4 individuals), followed by Irrawaddy dolphins (6.4 individuals), and finless porpoises (2.8 individuals).
5. Humpback dolphins exhibited a clustered distribution concentrated mainly in shallow estuarine waters (<10 m deep and <5 km from river mouths), whereas Irrawaddy dolphins were more widely distributed in farther coastal waters (<15 m deep and <15 km from river mouths), and finless porpoises were mostly found farthest from the shore in coastal waters (10–25 m deep and >15 km from river mouths).
6. The spatial distribution of the three cetaceans overlapped minimally, and this is likely to reflect the distribution of preferred prey resources, species interactions, and their differential responses to anthropogenic activities and species dominance.
7. The results from our study serve as baseline information for future research, conservation, and habitat management of these vulnerable and endangered coastal cetaceans. Conservation actions are recommended for the Matang area.

     

Production of a novel 9,12-dihydroxy-10(E)-eicosenoic acid from eicosenoic acid by Pseudomonas aeruginosa PR3

Microbial conversions of unsaturated fatty acids often generate polyhydroxy fatty acids, giving them new properties such as higher viscosity and reactivity. A bacterial strain Pseudomonas aeruginosa (PR3) has been intensively studied to produce mono-, di-, and trihydroxy fatty acids from different 9-cis-monoenoic fatty acids such as oleic acid, ricinoleic acid, and palmitoleic acid. However, from the results and the postulated similar metabolic pathways involved in these transformations, it was assumed that the enzyme system involved in transformation of the monoenoic fatty acid by strain PR3 could utilize fatty acids with different chain lengths and locations of the double bond. In this study was used as a substrate for bioconversion by strain PR3 eicosenoic acid (C20:1, ω-9) containing a singular cis double bond at different positions from the carboxyl end as oleic acid, and it was confirmed that PR3 could produce a novel 9,12-dihydroxy-10(E)-eicosenoic acid (DED) with 6.2% yield from eicosenoic acid. The structure of DED was confirmed using GC-MS, FTIR, and NMR analyses. DED production was maximized at 72 h after the substrate was added to the 24 h culture. Some other nutritional factors were also studied for optimal production of DED.     

Calcium oxalate accumulation and soil weathering in mats of the hypogeous fungus Hysterangium crassum

Fungal mats of Hysterangium crassum (Tul. and Tul.) Fischer occupied a mean of 9.6% of the upper 10 cm of soil developed under a 40–65 yr old stand of Douglas-fir (Pseudotsuga menziesii (Mirb.) Franco) in Oregon. This hypogeous basidiomycete exudes large amounts of oxalic acid, some of which precipitates with Ca in microscopic crystals of calcium oxalate, resulting in a mean CaC₂O₄ content of 82g m^?2 for the entire soil. Soil oxalate concentration was significantly greater within fungal mats (P < 0.01) and soil pH was significantly lower (P < 0.01) than in soil adjacent to mats. The quantity of Ca present as CaC₂O₄ is 0.5 the amount of exchangeable Ca in the soil and exceeds the mass of Ca lost annually in runoff. Scanning electron micrographs show intense chemical weathering, attributable to oxalate attack, in the immediate vicinity of hyphae. X-ray diffraction patterns of clay indicate bulk weathering is more intense within the fungal mats than in adjacent uncolonized soil.     

Eperythrozoon wenyonii infection in dairy cattle

Approximately 10 of 100 young heifers that had recently delivered their first calf--members of a large Colorado dairy herd--had a syndrome of swollen teats and distal portions of the hind limbs, prefemoral lymphadenopathy, transient fever, rough coat, decreased milk production, and subsequent weight loss and reproductive inefficiency. Acute clinical signs of disease were associated with large numbers of Eperythrozoon wenyonii seen on blood smears, and resolution of signs correlated with reduction or disappearance of the parasite. Other known causes of peripheral edema could not be documented. The parasite was transmitted to 4 of 7 nonlactating dairy cows destined to be culled and a splenectomized calf via IV inoculation of blood from parasitemic heifers, but clinical signs of infection were not induced.     

Characterization of turkey coronavirus from turkey poults with acute enteritis.

The present study was to characterize turkey coronavirus associated with turkey poult enteritis and mortality. Intestinal contents or intestines from affected turkey poults and inoculated turkey embryos contained coronaviruses as revealed by electron microscopy or were positive for turkey coronavirus by immunofluorescent antibody assay. Sucrose density gradient ultracentrifugation of the virus-containing intestinal homogenate yielded two opalescent bands corresponding to the buoyant densities of 1.14-1.15 and 1.18-1.20 g/ml, respectively. Coronaviral particles from intestinal contents or the sucrose density gradient preparation were mainly spherical in shape and had envelope and central depression. They were surrounded by a fringe of regularly spaced petal-shaped projections attached to the particles by a short stalk. Purified viruses hemagglutinated rabbit erythrocytes with a titer of 16. Major protein bands of purified viruses analyzed by SDS-PAGE were located at 200, 100-110, 50-60, and 30-35 kDa. The patterns of protein bands were consistent with those of Minnesota or Quebec turkey coronavirus isolates. A 568 bp nucleotide fragment of turkey coronavirus spike protein gene was amplified from RNA of inoculated turkey embryo intestine or purified virus. Sequence analysis of the 568 bp PCR product revealed high degree of identity with the corresponding spike protein gene sequence of human and bovine coronaviruses. The results indicated that turkey coronavirus was associated with turkey poults with acute enteritis.     

Identification of a novel Bacillus species isolated from Calculus Bovis

A novel Bacillus species of Calculus Bovis (cow gallstone) was isolated and identified in this study. Morphological features, bacterial fatty acid analysis using a microbial identification system, carbon source utilization profiles using Biolog system and 16S rDNA sequencing were employed to identify the isolated bacterium. This isolated bacterium was observed to be Gram‐negative, aerobic growing, rod‐shaped and short chain. The results of bacterial fatty acid analysis and physiological characteristics were not matched to the database. The main fatty acids found in the bacterium were 65.96% branched chain saturated fatty acids (iso C_11:0, anteiso C_11:0 and iso C_13:0～anteiso C_19:0). The bacterium oxidized 35 carbon sources and weakly responded with 49 of the 95 different carbon sources analyzed with the Biolog identification system. Based on 16S rDNA sequence analysis, this bacterium was classified as a novel Bacillus species.     

Seroepidemiology and genotyping of hepatitis E virus in Singapore reveal rise in number of cases and similarity of human strains to those detected in pig livers

Hepatitis E virus (HEV) causes 20 million infections worldwide yearly, of which only about 3.3 million are symptomatic. In developed Asian countries, HEV strains detected in human sera and in food sources were genetically similar, suggesting that indigenous HEV infections may be largely food‐borne. To assess the burden of hepatitis E in Singapore, we performed a seroepidemiologic study of the infection. Additionally, we carried out HEV genotyping on archived, residual HEV IgM‐positive serum samples collected between 2014 and 2016 (n = 449), and on pig liver samples (n = 36) purchased from wet markets and supermarkets. Our study shows a rise in hepatitis E incidence (IgM) from 1.7 to 4.1 cases per 100,000 resident population from 2012 to 2016 and an increase in hepatitis E IgG positivity rate among residents from 14% in 2007 to 35% in 2016. Other findings also suggest the epidemiology of hepatitis E in Singapore has shifted, from it being mainly a disease imported from the Indian subcontinent, to one that is now increasingly prevalent in our resident population. Genotypes obtained from 143 human samples identified the majority to be genotype 3 (n = 121), 21 to be genotype 1 and one to be genotype 4. Further phylogenetic analyses suggest genotype 3a to be the cause of indigenous infections in residents, which showed genetic similarity to the genotype 3a strains detected in pig livers. This link between the strains in the majority of human samples and those in pig livers consumed by the public suggests a possible food‐borne route of HEV infection in Singapore.