Digestion of rice straw and oil palm fronds by microflora from rumen and termite bacteria, in vitro

The digestion and Volatile Fatty Acid (VFA) production from rice straw and oil palm fronds by cellulolytic bacteria isolated from the termite Coptotermes curvignathus were investigated. The bacteria were Acinetobacter strain Raminalimon, Enterobacter aerogenes strain Razmin C, Enterobacter cloacae strain Razmin B, Bacillus cereus strain Razmin A and Chryseobacterium kwangyangense strain Cb. Acinetobacter strain Raminalimon is an aerobic bacterium, while the other species are facultative anaerobes. There were significant differences (p<0.05) among the bacteria for Dry Matter (DM) lost and acetic acid production from rice straw and Acinetobacter strain Raminalimon showed the highest activity. The facultative bacteria C. kwangyangense strain Cb (cfu mL(-1) 231 x 10(-6), OD: 0.5), E. cloacae (cfu mL(-1) 68 x 10(-7), OD: 0.5) and E. aerogenes (cfu mL(-1) 33 x 10(-7), OD: 0.5) were used for digestion study with the rumen fluid microflora. The in vitro gas production technique was applied for the comparative study and the parameters measured were pH, gas (volume), dry matter lost, acetic acid, propionic acid and butyric acid concentrations. pH was not significantly (p<0.05) different among the five treatments. The bacterium C. kwangyangense strain Cb showed the highest activity (p<0.05) for DM lost, acetic acid, propionic acid and butyric acid production from rice straw when compared to the other bacterial activities. There was no significance (p<0.05) difference between the three bacteria for the dry matter lost of oil palm fronds but the production of Volatile Fatty Acids (VFA) was significantly (p<0.05) high in the treatment which was inoculated with C. kwangyangense strain Cb. The Gen Bank NCBI/EMBL accession numbers for the bacterial strains are EU332791, EU305608, EU305609, EU294508 and EU169201.     

Phytoplasmas associated with disease of coconut in Malaysia: phylogenetic groups and host plant species

Coconut palm ( Cocos nucifera ), oil palm ( Elaeis guineensis ), Bermudagrass ( Cynodon dactylon ) and Madagascar periwinkle ( Catharanthus roseus ) with symptoms indicative of phytoplasma disease were collected from different locations in Malaysia. PCR assays employing phytoplasma universal rRNA gene primers P1/P7 alone or P1/P7 followed by R16F2n/R16R2 detected phytoplasmas in eight out of 20 Malayan Red Dwarf (MRD), nine out of 12 Malayan Yellow Dwarf (MYD) and 12 out of 12 Malayan Tall (MT) coconut palms displaying coconut yellow decline symptoms. Positive detections were also obtained from six out of six oil palm seedlings showing symptoms of yellowing and necrosis, from 10 out of 10 Bermudagrass samples with white leaf symptoms, and from eight out of eight periwinkle plants showing phyllody, virescence, little leaf, proliferation and foliar yellowing. Phytoplasmas were not detected in any of the symptomless plants tested. Sequencing and phylogenetic analysis of PCR products determined that phytoplasmas infecting both MRD and MT coconuts and Bermudagrass in Serdang, Selangor State, were all members of the 16SrXIV ' Candidatus Phytoplasma cynodontis' group, whereas isolates in periwinkle in Serdang were all members of the 16SrI ' Ca. Phytoplasma asteris' group. However, the phytoplasmas detected in MYD coconuts and oil palms from Banting, Selangor State, and in periwinkle from Putrajaya were collectively very similar (99%), but shared <97·5% similarity with 16S rDNA sequences of all other known phytoplasmas, indicating that they represent a novel taxonomic group. Thus, at least two phylogenetically distinct phytoplasmas are associated with the coconut yellow decline syndrome in Malaysia, both of which were also detected in other plant species.     

Characterization of <Emphasis Type="Italic">Pectobacterium carotovorum</Emphasis> causing a new soft rot disease on okra in Malaysia

Dark brown, necrotic pods with extensive water-soaked lesions caused by plant pathogenic bacteria were found on okra plants in different fields in Malaysia in 2010. PCR amplification of the pectate lyase (pel) gene and amplification of the 16S–23S rRNA (ITS) with G₁ and L₁ primers produced 434-, 535- and 570-bp fragments, respectively. From the similarity between the results of biochemical tests and their equivalency with standard bacteriological sources, PCR-based pel gene, and RFLP analysis of the ITS-PCR products, all isolates were identified as Pectobacterium carotovorum. This is the first report of P. carotovorum in okra from Malaysia.     

Cross‐infectivity of oil palm by Phytophthora spp. isolated from perennial crops in Malaysia

Bud rot disease or “Pudricion del cogollo” (PC) of oil palm is a major constraint on production in Colombia and neighbouring countries such as Brazil, Costa Rica, Ecuador, Nicaragua, Panama, Peru and Surinam. To date, there are no documented reports of Phytophthora disease of oil palm in South‐East Asia. This research, therefore, was conducted to determine the pathogenic potential of Phytophthora palmivora and Phytophthora nicotianae on oil palm using both in vitro and nursery inoculation experiments. In vitro inoculation of both P. palmivora and P. nicotianae on immature oil palm leaflets caused discoloration within 2 days of inoculation and incubation at 25 ± 1.5°C, 100% RH. Similarly, in nursery trials, lesions formed on the buds (unopened leaflets) 3 days after inoculation with P. palmivora or P. nicotianae zoospore suspensions. No lesions developed on untreated leaflets in either in vitro or nursery inoculation experiments. Phytophthora spp. were re‐isolated from leaflet lesions and confirmed as the inoculated pathogens.     

Influence of sheep grazing on soil chemical properties and growth of rubber (Hevea brasiliensis) in Malaysia

In a rubber estate in Kelantan, Malaysia, sheep grazing increased N, P, Ca and Mg levels and soil pH. Foliar N, P, Ca, Mg and Na also increased with grazing, but K decreased. The girth of rubber trees under grazing had a higher increment than in the ungrazed areas.     

Potential use of selected citrus rootstocks and interstocks against HLB disease in Malaysia

Huanglongbing (HLB), also known as citrus greening disease, has spread to and is seriously threatening the citrus industry in more than 40 countries. The HLB pathogen is a phloem limited bacteria, Candidatus Liberibacter spp. There is little information pertaining to the effects of different citrus rootstocks and interstocks against HLB in Citrus reticulata. The objective of this study was to evaluate the beneficial effects of different combinations of citrus rootstocks and interstocks against HLB using side grafting. There were no symptoms of HLB when C. grandis was used as rootstock with C. hystrix as the interstock and vice versa six months after inoculation. However, C. Liberbribacter asiaticus was detected in the scion using second PCR amplification. A high rate of disease severity was observed when C. aurantum was used as rootstock and C. aurantifolia as the interstock and vice versa. This study showed that C. Liberibacter asiaticus can be detected by conventional PCR and characteristics of their detrimental effects include low rate of vegetative growth and reduction of dry matter, root dry matter, plant height and stem diameter.     

The study of seroprevalence of hepatitis E virus and an investigation into the lifestyle behaviours of the aborigines in Malaysia

Malaysia is a non‐endemic country for hepatitis E virus (HEV) infection. However, seroprevalence as high as 50% among samples of aboriginal people were reported over two decades ago. A total of 207 samples collected from seven aboriginal villages in rural settlements across two states in Malaysia were analysed for anti‐HEV IgG and IgM by an enzyme‐linked immunoassay. Following the detection of anti‐HEV seroprevalence, we organized health outreach to inform and educate the community. Qualitative interviews were conducted with individuals tested positive for anti‐HEV antibodies. Data derived from interviews and observations were used to investigate possible lifestyle behaviours associated with HEV infection. Anti‐HEV IgG was detected in six samples (5.9%) from the village of Dusun Kubur. Qualitative inquiry and observation study revealed poor dietary and household hygiene, contaminated food and water, contact with animal faeces, unsanitary and domestic waste disposal, and wildlife reservoirs could be the contributing factors for transmission and acquisition of HEV infection. Investigation during health outreach is important to provide insights for future empirical research and implementation for improvement of lifestyle behaviours among the aborigines. Managing the risk of HEV infection in the aborigines may reduce the risk of HEV transmission to the local communities.     

Characterization of Malaysian Pectobacterium spp. from vegetables using biochemical, molecular and phylogenetic methods

In August 2011, vegetable crops showing symptoms of maceration and water soaked lesions on their tuber, leaf, and fruit were collected from four major vegetable growing states in Malaysia including Pahang, Johor, Melaka and Selangor. The majority of the causal organisms isolated from infected tissues (52 strains) were identified as Pectobacterium spp. based on PCR amplification of the pectate lyase (pel) gene and amplification of the 16S-23S rRNA (ITS) with G1 and L1 primers. Physiological and biochemical assays divided Malaysian Pectobacterium species into two main groups: Pectobacterium wasabiae and Pectobacterium carotovorum subsp carotovorum. Partial sequence of PCR product from reaction of putative Pectobacterium spp. with 16S rRNA confirmed the results obtained from physiological and biochemical assays used for identification of the bacterium. Application of specific primers such as Eca1F/Eca2r, Br1f/L1r, EXPCCF/EXPCCR, and also ITS-PCR following by RFLP by restriction enzyme (RsaI) successfully differentiated Malaysian P. wasabiae and P. carotovorum subsp carotovorum isolates from other species and subspecies of Pectobacterium. Phylogenetic analysis of Malaysian isolates with housekeeping genes (mdh, gapA) grouped Malaysian P. carotovorum subsp carotovorum and P. wasabiae in the same cluster with P. carotovorum subsp carotovorum (Ecc380) and P. wasabiae (SCRI488) respectively.