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Microbially induced carbonate precipitation by ureolysis is a biomineralization process that has been adapted by various microorganisms in different natural environments. This widespread natural phenomenon can be employed in numerous civil engineering and soil stabilization applications. In the present study, the potential of indigenous soil urease-producing bacteria as potential agents for soil stabilization methods was investigated. Assessment of the eight active urease-producing bacterial species isolated from the farm soil samples has demonstrated that all the isolates were Gram-positive rod-shaped bacteria with promising characteristics such as the formation of endospore which is essential for bacterial survival in harsh conditions within the soil environment. The pH profile and growth profile of the isolates were studied and urease activity was measured by the phenol hypochlorite assay method. Two isolates designated isolate O6w and isolate O3a were selected based on the highest urease activity recorded at 665 U/mL and 620 U/mL, respectively, and they were able to increase and sustain alkaline culture condition (pH 8.71 ± 0.01 and 8.55 ± 0.01) which was suitable for CaCO3 precipitation. The isolates were identified based on 16S ribosomal RNA sequencing to be Bacillus cereus (O6w) and Bacillus paramycoides (O3a). This current study suggested that indigenous soil ureolytic bacteria are potential raw material for the biotreatment of soils stability.

Fusarium fruit rot caused by fusarium species pathogens can be considered a threat to economic loss. The use of synthetic antifungals to control the pathogens has failed to the development of resistance of pathogens. Previous studies reported that plant extracts may contain various bioactive constituents that are able to control pathogen growth. Hence, the aims of this study are to examine the inhibition activity of some plant fresh extracts on mycelial growth and morphological changes of Fusarium oxysporum, F. proliferatum, and f. solani, causal agents of Fusarium fruit rot. Aqueous extract of selected plants was evaluated for their inhibition activity against all the three fusarium pathogens under in vitro condition using poisoned food bioassay. Averrhoa bilimbi fruit extract demonstrated a highly significant effect against mycelial growth of the pathogens with fungal inhibition percentage of 80.51% for F. oxysporum, 61.28% for F. proliferatum and 58.97% for f. solani. The results showed that the highest formation of 100% extract has lowered the conidia concentration and germination percentage of F. oxysporum (35.43%), F. proliferatum (47.61%), and f. solani (38.67%) compared to the control. Significant shriveled mycelia were observed via scanning electron microscope (SEM) on the pathogens treated with a. bilimbi extract, indicating morphological changes occurred in the cell membrane compared to the control in which the mycelia were in normal form. This innovation, which can be prepared and applied at home, has the potential as an eco-friendly and a benign approach to control fruit rot pathogen.

The rising concern regarding the negative impact of synthetic pesticides has led to the search for alternative means of pest control. Vermicomposting the mixture of oil palm empty fruit bunch and neem (Azadirachta indica) leaves, with the latter known to have pesticidal value, is therefore of great interest and significance to be studied. The present study was conducted to evaluate the chitinase, protease and β-1,3-glucanase activity of neem leaf-based vermicompost as an indication of its biocontrol properties. The total microbial population of different composition of the vermicompost was also investigated. The results showed that at 10% neem composition, an increment in microbial population, chitinase and protease activities was observed in the end product. A higher concentration of neem exerted a suppressive effect on the microbial population as well as enzymatic activity. This study suggested that the addition of an appropriate composition of neem leaves as one of the raw materials for vermicomposting would potentially enhance the performance of vermicompost as biofertilizer as well as biopesticide.

Crepidium amplectens and Crepidium lowii are reported as two newly recorded Crepidium species belong to sect. Commelinodes from Peninsular Malaysia. The taxonomic affinity is discussed and validated involving another two of their complexes, Crepidium micranthum and Crepidium rheedei subsp. rheedei, to which they share some similar characteristics of the leaves and flowers. The comparative study was conducted based on a comprehensive description and clear photographic evidence of the macro-morphology and surface micro-morphology of the labellar and gymnostemium parts. This paper emphasizes the usefulness of diversity and distribution of ultrastructures and epicuticular sculpturing of the reproductive parts in delimiting the confusable and sympatric Crepidium species, and a brief note on the nectar or reward secretion in the genus with inference on the ecological adaptation and pollination strategies. Macro-morphologically, C. amplectens differs from C. lowii in having the leaves smaller, greenish and widely ovate. Individually, both species can be differentiated from C. micranthum based on the smaller flower and shorter labellum auricles, and inconspicuous gymnostemium stelidia. Micro-morphologically, C. amplectens is also distinguishable from C. lowii by having different diversity and distribution of the epicuticular ornamentation and papillae on the labellum and gymnostemium. Also, amongst the four studied species, only C. lowii has nectarostomata, occurring on the fovea or nectary.

Background: Rural and urban people in Nigeria made use of medicinal plants as their curative measures, based on their ancient belief that propounded the authenticity of these plants in treating ailments. Materials and methods: Data were collected through semi-structured questionnaires across the study area (3senatorial districts),The collected plant species were authenticated and given voucher numbers, and the demographic data was subjected to Chi-square (x2) comparisons using SPSS version 22. Results: The major family among the surveyed plants, were Fabaceae (dominant), followed by Anacardiaceae and Combretaceae. Moreover, out of 97 respondents, 81 were male (83.5%) and 16 were female (16.5%). It was observed that respondents that were 41-50 years were many into practice ( p = 0.13. The majority of the respondents as compared using chi-square across the parameters, were illiterates (p = 0.06), and very few had a formal training or exposed to workshops (p = 0.02), while the majority inherited and utilized herbal medicine practice as their main sources of income (p = 0.04). The fidelity levels ranged as follows: For diarrhea (18.5% -100%), dysentery (11.11- 45%), pile (11.11 – 50%) and, ulcer covered 9.1% -100%. The ailments were in the range of 0.69 – 0.75 factors of informant consensus. Plant species with RFC values of 0.34, 0.27 and 0.21 as well as those with 0.1, were regarded to have the highest RFC values. Conclusion: Information collected were mainly on cases of gastrointestinal ailments and first of its kind on the use of medicinal plants in Yobe State, Nigeria.

Metabolic changes in plants may serve as an early signal for infection in response to pathogen intrusions. In this study, metabolomic fingerprinting analysis was conducted on banana plantlets inocu...

Gallic acid and catechin are the most abundant phenolic and flavonoid contents found in all plant extracts. The contents and the bioassay-guided fractionating substances of the Sclerocarya birrea (A. Rich) Hochst (Anacardiaceae) fraction played vital roles. The goals of the study were to determine the contents of some useful medicinal plants and the bioassay-guided fractionation substances of S. birrea fraction compounds capable of acting against Salmonella isolate using LC-MS/LC-HRMS (Dionex ultimate 3000 RS UPLC with Thermo Scientific Q Exactive Orbitrap Hybrid Tandem Mass Spectrometer). The Folin–Ciocalteu reagent procedure and flavonoid content determination were conducted spectrophotometrically. Bioassay-guided fractionation, chronological partitioning, and screening of the antibacterial action against Salmonella typhi were performed. The ethyl acetate fraction extracts of S. birrea stem (bark) extract were analyzed using LC-MS/LC-HRMS. The gallic acid content increased tremendously in Vachellia nilotica (L.) P.J.H. Hurter and Mabb (Fabaceae) pod extracts with curve fitting (R2 = 0.9958). Catechin content increase was significantly increased in S. birrea stem (bark) extracts followed by that of V. nilotica pod extracts with curve fitting (R2 = 0.9993); they were all significantly different in the Guiera senegalensis J.F. Gmel. and the Leptadenia lanceolata (Poir.) Goyder leaves extracts at p value S. birrea stem (bark) ethyl acetate fraction extract was the MIC, where no MBC was recorded and susceptible to the positive control with the highest inhibition zone, followed by the ethyl acetate fraction extract at 10 mg/ml (9.7 ± 0.0) at Turkey’s p < 0.0001. Vidarabine is one of the novel compounds, specifically having antimicrobial actions, found in the S. birrea stem (bark). Reasonable amounts of phenolic and flavonoid contents determined the actions of the individual plant extract.

Scurrula ferruginea is a type of mistletoe from the Loranthaceae family found in Southeast Asia and distributed in tropical regions, is known to have healing effects for many disorders. Preservation of the bioactive metabolites is dependent on the sample preparation and drying methods as well as extraction solvents used. Thus, the present study was carried out to investigate the effect of different drying methods and extraction solvents on the total phenolic and the antioxidant activities of S. ferruginea leaves. The leaves of S. ferruginea were air and oven (60°C) dried and extracted with aqueous, organic and aqueous-organic solvents. The total phenolic content (TPC) and total flavonoid content (TFC) were used to gauge the phenolic antioxidants extracted, while the antioxidant activity was evaluated by measuring the scavenging effect on 2,2- diphenyl-1-picrylhydrazyl (DPPH) radical and via the ferric reducing antioxidant power (FRAP) assay. The optimum conditions obtained were oven drying, 80% acetone with values of 3.47 mg/10 g DW (extract yield), 171.29 mg GAE/1 g DW (TPC), 31.91 mg QE/1 g DW (TFC), 93.2% (DPPH), 26.41 mg TE/1 g DW (FRAP) and 7.41 μg/mL (IC50). The present findings suggested that sample preparation variability using different drying methods and solvents for extraction play a crucial role in enhancing the efficiency of extraction and efficacy of the antioxidant compounds in S. ferruginea leaf extracts.

Plants play vital roles in many health care systems, be it rural or an urban community. Plants became familiar as medicine due to the primordial ideologies and believed. Several plant parts served as medicines to so many ailments including gastrointestinal ailments, due to the fact that their active ingredients are powerful against the microbes as well as healing so many physiological abnormalities. The principal antimicrobial components were used to inhibit the growth of microbes (S. aureus, E. coli, Salmonella spp, B. cereus, and B. subtili,), as well as most of the recognized compounds in most plants were aromatic or saturated organic compounds which enabled the plants to be active against the gastrointestinal microbes. The commonly used diluents were; water, methanol and Di methyl sulphate oxides to ascertain the level of activity of the plants. As such, plant materials in one way or the other are very active when dealing with microbes due to their active ingredients or the phytoconstituents. Most of the microbes identified in many reviewed researches were enteric bacterial species, by which divided into both gram negative and gram positive bacterial isolates, they differ in their cell components, which are the main targets of bioactive constituents to deal with any bacteria. However, certain parasites contributed towards the production of ailments for their survival and causing havoc to the hosts and sometimes be mutualistic.

The soil environment is teeming with living organisms full of biological life and is one of the most abundant and diverse ecosystems on earth. However, due to unsustainable agriculture practices, the fertile topsoil is often lost; the soil becomes useless for agriculture. Therefore, we cannot lose sight of the fundamental role of the soil biological entities in maintaining the sustainability of the soil, and the role of organism’s biodiversity in the soil to counter against various stress. Therefore, the main objective of this work is detecting diversity and density of soil invertebrates between a cultivated field (Ladang 2) and uncultivated field (CEFS), Results showed that the highest abundance of invertebrates was recorded at CEFS 834 with 3.7 m2 density and the lowest was in Ladang 2: 225 with 1.2 m2 density, belonging to 35 different morphospecies from 4 Phylum, 4 subphyla, 6 class, 2 subclass, and 19 orders. A total of 15 morphospecies were collected in the cultivated field Ladang 2, while 25 morphospecies were collected in the uncultivated filed CEFS. Out of 25 morphospecies found in this area, 10 were exclusive to this environment. Annelida, Amphipoda, Isopoda, Collembola, and Hymenoptera were the dominant taxa in CEFS, comprising: 8.15%, 10.55%, 17.98%, 14.14%, and 26.97% respectively. But Acarina, Coleoptera, and Insect larva 39.68%, 5.95%, and 3.97% respectively were the dominant taxa in Ladang 2. The diversity was highest in the CEFS (H = 2.29) and lowest in the Ladang2 field (H = 1.74). Unfortunately, many aspects of the soil invertebrates have not been documented in Malaysia. For example, why they become dominant in a certain location and what is the impact on that particular ecosystem. This study showed that it is essential to maintain natural habitat because agricultural practices have a major influence on soil invertebrate diversity and hopefully through this study able to suggest sound agriculture management to safeguard the soil ecosystems. Due to the limited information about soil invertebrates in Malaysia and the limited scope of this study, it is premature to make any conclusion and therefore warrant further studies.

The floral micromorphology of critically endangered Paphiopedilum Pfitzer [P. barbatum (Lindl.) Pfitzer, P. callosum var. sublaeve (Rchb.f.) P.J.Cribb and P.niveum(Rchb.f.) Stein] were analyzed concerning either infrageneric taxonomy or physioecological demands. The first two species are phylogenetically close and superficially identical but occur with distinct phytogeographical distributions in the region. The third species is a phylogenetically distant congener that inhabits limestone areas in the northern part of Peninsular Malaysia. Using scanning electron microscopy (SEM), we investigated the surface of the dorsal sepal, synsepal, lateral petals, pouch or labellum, and staminode. Amongst the investigated features were epicuticular waxes, epicuticular ornamentation, trichome distribution and type, pustular glands, and papillae. Our study supports the distinction of P. barbatum from P. callosum var. sublaeve, which belong to subgenus Paphiopedilum, and from P. niveum, a species belonging to subgenus Brachypetalum, a separated monophyletic clade. Comparatively, P. barbatum has Type III non-glandular trichomes on the margin of its lateral petals, which are absent in P. callosum var. sublaeve. Paphiopedilum callosum var. sublaeve and P. niveum are distinguishable from P. barbatum by a confined distribution of papillae. The epicuticular ornamentation and distribution of trichomes on staminode discriminate between P. barbatum and P. callosum var. sublaeve and differentiates them from P. niveum. Compared to P. barbatum and P. niveum, stomata in P. callosum var. sublaeve were superficial with prominently raised guard cells. From the physioecological view, the absence of glandular trichomes, and the low occurrence of papillae and stomata on the floral parts explain the unscented flowers of P. barbatum and P. callosum var. sublaeve. A combination of the features examined is taxonomically valuable for delimitation of the species at the infrageneric level, although the diagnostic characters are far inadequate for a generic taxonomic revision. A study with a more extensive sampling from the three subgenera of Paphiopedilum, including subgenus Parvisepalum, is anticipated to elucidate the level of variation of the analyzed microcharacters. Key Words: physioecological importance, Peninsular Malaysia, subgenus Brachypetalum, subgenus Paphiopedilum, scanning electron microscopy, taxonomic delimitation Abstract. The floral micromorphology of critically endangered Paphiopedilum Pfitzer [P. barbatum (Lindl.) Pfitzer, P. callosum var. sublaeve (Rchb.f.) P.J.Cribb and P.niveum(Rchb.f.) Stein] were analyzed concerning either infrageneric taxonomy or physioecological demands. The first two species are phylogenetically close and superficially identical but occur with distinct phytogeographical distributions in the region. The third species is a phylogenetically distant congener that inhabits limestone areas in the northern part of Peninsular Malaysia. Using scanning electron microscopy (SEM), we investigated the surface of the dorsal sepal, synsepal, lateral petals, pouch or labellum, and staminode. Amongst the investigated features were epicuticular waxes, epicuticular ornamentation, trichome distribution and type, pustular glands, and papillae. Our study supports the distinction of P. barbatum from P. callosum var. sublaeve, which belong to subgenus Paphiopedilum, and from P. niveum, a species belonging to subgenus Brachypetalum, a separated monophyletic clade. Comparatively, P. barbatum has Type III non-glandular trichomes on the margin of its lateral petals, which are absent in P. callosum var. sublaeve. Paphiopedilum callosum var. sublaeve and P. niveum are distinguishable from P. barbatum by a confined distribution of papillae. The epicuticular ornamentation and distribution of trichomes on staminode discriminate between P. barbatum and P. callosum var. sublaeve and differentiates them from P. niveum. Compared to P. barbatum and P. niveum, stomata in P. callosum var. sublaeve were superficial with prominently raised guard cells. From the physioecological view, the absence of glandular trichomes, and the low occurrence of papillae and stomata on the floral parts explain the unscented flowers of P. barbatum and P. callosum var. sublaeve. A combination of the features examined is taxonomically valuable for delimitation of the species at the infrageneric level, although the diagnostic characters are far inadequate for a generic taxonomic revision. A study with a more extensive sampling from the three subgenera of Paphiopedilum, including subgenus Parvisepalum, is anticipated to elucidate the level of variation of the analyzed microcharacters. Key Words: physioecological importance, Peninsular Malaysia, subgenus Brachypetalum, subgenus Paphiopedilum, scanning electron microscopy, taxonomic delimitation

Diuron is an alternative biocide suggested to replace organotin in formulating antifouling paints to be applied on water-going vessels hull. However, it is potentially harmful to various non-targeted marine organisms due to its toxic properties. Present study aimed to isolate, screen and identify the potential of Diuron-degrading bacteria collected from the marine sediments of Port Klang, Malaysia. Preliminary screening was conducted by exposing isolated bacteria to 430ng/L (background level), followed by 600ng/L and 1000ng/L of Diuron concentrations. Nine bacteria colonies survived the exposure of the above concentrations. However, only two strains can tolerate to survive up to 1000μg/L, which were then characterised and identified using phenotypic tests and the standard 16S rRNA molecular identification. The strains were identified as Comamonas jiangduensis SZZ 10 and Bacillus aerius SZZ 19 (GenBank accession numbers: KU942479 and KU942480, respectively). Both strains have the potential of Diuron biodegradation for future use.

Chitinase 42 kDa produced by Trichoderma harzianum has been proven as a prime compound to be excreted onto the hyphae of the pathogen causing localised cell wall lysis at the point of interaction. This finally initiate the process of the host cell becomes empty of cytoplasm, disintegrates and shows a rapid collapse. This study investigates the existence of N-glycan linked mannose in chitinase 42 kDa produced by the Malaysian T. harzianum strain BIO10671. The chitinase 42 kDa from T. harzianum BIO10671 was initially purified using anion exchange chromatography prior to a series of experiments such as immunoblotting against the chitinase 42 kDa antibody, lectin staining for detecting any terminal linked mannose, and galactofuranose detection to determine the presence of galatofuranose components in glycoproteins. The enzyme purification harvested about 12-fold of chitinase 42 kDa from T. harzianum BIO10671 with strong indication of the presence chitinase 42 kDa presence on SDS-Page. This was confirmed by immunoblotting with a strong response around 42 kDa after overnight incubation in chitinase 42 kDa antibody suggesting that the gene for chitinase 42 kDa was greatly expressed in this strain. There are no intervation of galatofuranose on any of the terminal mannose in chitinase 42 kDa as shown by negative results on samples treated with or without endoglycosidase-H and lectin staining. Therefore, it can be concludeed that glycosylation occurred in the chitinase 42 kDa from T. harzianum 42 kDa was not in the form of N-glycan linked mannose as expected.

Graphical abstract, Polycyclic Aromatic Hydrocarbons (PAHs) are complex and widely distributed environmental pollutants that can affect living ecosystems. This study was conducted to rapidly degrade phenanthrene and pyrene representing low and high molecular weight of PAHs, respectively. Cronobacter sakazakii MM045 (KT933253) was identified from used engine oil of contaminated soil. PAHs biodegradation was carried out using 2,6-dichlorophenol indophenol (DCPIP) assay. Biodegradation influencing factors including agitation, temperature, pH, inoculums volume and salinity were enhanced using Response Surface Methodology (RSM) by Central Composite Design (CCD). Phenanthrene and pyrene biodegrading metabolites were identified using gas chromatography mass spectrophotometer (GCMS). • Initial biodegradation indicated 75.2% and 54.3% phenanthrene and pyrene degraded by C. sakazakii MM045 within 24 h. After CCD optimisation, 100% degradation was achieved for each of the phenanthrene and pyrene, resulting in the formation of intermediate metabolites. • The identified phenanthrene metabolites were 3,4-dihydroxyphenathrene, phthalic acid, pyruvic acid, acetic acid and oxalic acid. Pyrene intermediates comprised pyrene cis-4,5-dihydrodiol, 3,4-dihydroxyphenanthrene, phthalic acid, pyruvic acid, acetic acid and lactic acid. • Cronbacter sakazakii MM045 was proven to be rapid and effective in degrading PAHs within 24 h despite the unavailability of existing literatures on PAHs biodegradation.

Background Vibriosis is an important bacterial disease of cultured marine fishes worldwide. However, information on the virulence and antibiotic resistance of Vibrio spp. isolated from fish are scarce. This study investigates the distribution of virulence associated genes and antibiotic resistance patterns of Vibrio spp. isolated from cage-cultured marine fishes in Malaysia. Results A total of 63 Vibrio spp. isolated from 62 cultured marine fishes in various geographical regions in Peninsular Malaysia were analysed. Forty-two of the isolates (66.7%) were positive for all chiA, luxR and vhpA, the virulence genes produced by pathogenic V. harveyi. A total of 62 Vibrio isolates (98%) had tlh gene of V. parahaemolyticus, while flaC gene of V. anguillarum was detected in 43 of isolates (68%). Other virulence genes, including tdh, trh, hlyA and toxR vc were absent from any of the isolates. Multiple antibiotic resistance (MAR) was exhibited in all strains of Harveyi clade, particularly against ampicillin, penicillin, polypeptides, cephems and streptomycin. The MAR index ranged between 0.06 and 0.56, and 75% of the isolates have MAR index of higher than 0.20. Host species and geographical origin showed no correlation with the presence of virulence genes and the antibiotic resistance patterns of Vibrio spp. Conclusions The study indicates that majority of Vibrio spp. isolated from cultured marine fishes possess virulence genes, but were not associated with human pathogen. However, the antibiotics resistance is a real concern and warrants ongoing surveillance. These findings represent an updated knowledge on the risk of Vibrio spp. to human health, and also provides valuable insight on alternative approaches to combat vibriosis in cultured fish.

Blood disease is a destructive bacterial infection that causes severe yield loss to the banana industry. Ideally, an environmental friendly yet practical approach is necessitated in the search for effective treatment against the disease. Endogeic earthworms are soil biota that help in improving soil physico-chemical and biological properties and thuspromote plant health. The present study assessed the effect of Pontoscolex corethrurus on banana infected by blood disease. The effect of earthworms was evaluated through observations on stem and root morphology, anatomy and total phenolic contents (TPC). P. corethrurus was inoculated into polybags planted with banana plantlets; inoculation of blood disease bacterium (BDB) into the roots of the plantlets was done using a drenching method. Stem and root sections of the plantlets were preserved in FAA for histological study upon harvest. The remaining sections were freeze-dried for TPC analysis. Root sections of plantlets infected with BDB showed lower root biomass compared to the control and earthworm-inoculated plantlets. Observations under microscope also showed that tissue necrosis at the vascular bundles of infected roots were more severe compared to the earthworm-inoculated and unaffected plantlets. Plantlets inoculated with earthworms had the highest root TPC, followed by the unaffected plantlets and plantlets infected with BDB. Although infected plantlets with earthworm inoculation showed disease symptoms, the disease severity was slightly less. The results suggested the potential role of P. corethrurus in improving plant health for disease management and sustainable agriculture.

Blood disease is a wilt disease that greatly affects the banana cultivation. Besides chemicals, the utilization of natural soil-biological entity may provide an alternative in searching for effective and environmental friendly approach against the disease. Endogeic earthworms are known as soil-engineers that modify soil properties that may favor or suppress different microbial populations. In the study, the effect of endogeic earthworm, Pontoscolex corethrurus on blood disease bacterium (BDB) was evaluated through in-vitro study. Earthworm casts, mucus and worm-worked soil collected were inoculated onto tetrazolium chloride (TZC) medium seeded with BDB at different concentrations. Molecular identification using 16S rRNA primers was then performed to identify the microbial isolates that inhibited BDB. Phylogenetic tree was constructed to determine the similarity among the isolates with Pseudomonas sp. sequences deposited in the GenBank. The results showed that bacterial colonies from earthworm mucus inhibited BDB at 103 and 105 cell/ml. However, no inhibition was observed at higher BDB concentration (107 cell/ml). Similarly, earthworm cast and worm-worked soil did not show inhibition towards BDB. Bacterial isolates obtained from earthworm mucus were classified into the genus Pseudomonas. The study suggested the potential role of P. corethrurus in enhancing the growth of beneficial microorganism in remediating blood disease.

A possible tributyltin (TBT)-degrading bacterium isolated from contaminated surface sediment was successfully identified as Klebsiella sp. FIRD 2. It was found to be the best isolate capable of resisting TBT at a concentration of 1000 μg L(-1). This was a concentration above the reported contaminated level at the sampling station, 790 μg L(-1). Further studies revealed that the isolate was Gram negative and resisted TBT concentrations of up to 1500 μg L(-1) in a Minimal Salt Broth without the addition of any carbon source within the first 48 h of incubation. It is expected that additional work could be conducted to check the degradation activity of this new isolate and possibly improve the degradation capacity in order to contribute to finding a safe and sustainable remediation solution of TBT contamination.

Streptococcus agalactiae is one of the most important pathogens infecting tilapia worldwide. In this study, we determined the serotype, sequence type (ST), virulence gene profile and pathogenicity of S. agalactiae isolated from cultured fish. A collection of 256 S. agalactiae that were previously isolated from outbreaks of streptococcosis in red hybrid tilapia in Peninsular Malaysia were studied. The genotype and ST of the isolates were determined by molecular serotyping of the capsular polysaccharide (cps) gene clusters, and multi-locus sequence typing (MLST). The virulence gene profiles were constructed using m-PCR for 14 standard virulence genes with published primers. Two serotypes, with different associated STs, were identified in this study: serotypes Ia ST7 and III ST283. Serotype Ia ST7 lacks lmb, scpB, pavA, fbsB, cyl, bca, cspA and bac genes, which were present in serotype III ST283. The LD50–240h of S. agalactiae in red hybrid tilapia for serotypes Ia ST7 and III ST283 were 8.7 × 103 CFU/mL and 6.3 × 103 CFU/mL, respectively. The histopathological findings were consistent with meningitis, necrosis of hepatocytes, coagulative tubular necrosis and hypocellular of the spleen. Histopathological scores revealed significant differences (p

This study investigated the environmental factors associated with the presence of Vibrionaceae in economically important cage-cultured tropical marine fishes: the Asian Seabass Lates calcarifer, snapper Lutjanus sp., and hybrid grouper Epinephelus sp. Fish sampling was conducted at monthly intervals between December 2016 and August 2017. The body weight and length of individual fish were measured, and the skin, eye, liver, and kidney were sampled for bacterial isolation and identification. Water physicochemical parameters during the sampling activities were determined, and the enumeration of total Vibrionaceae count was also conducted from water and sediment samples. Nine species of Vibrio were identified, including V. alginolyticus, V. diabolicus, V. harveyi, V. campbellii, V. parahaemolyticus, V. rotiferianus, V. furnissii, V. fluvialis, and V. vulnificus. Photobacterium damselae subsp. damselae was also identified. A total of 73% of the isolated Vibrio belonged to the Harveyi clade, followed by the Vulnificus clade (5.5%) and Cholera clade (0.6%). Highest occurrence of Vibrio spp. and P. damselae subsp. damselae was found in hybrid grouper (72%), followed by Asian Seabass (48%) and snapper (36%). The associations of Vibrio spp. and P. damselae subsp. damselae with the host fish were not species specific. However, fish mortality and fish size showed strong associations with the presence of some Vibrio spp. On average, 60% of the infected cultured fish exhibited at least one clinical sign. Nevertheless, inconsistent associations were observed between the pathogens and water quality. The yearlong occurrence and abundance of Vibrionaceae in the environmental components indicate that they might serve as reservoirs of these pathogens.

Bacterial wilt disease has caused a serious threat to the banana industry in Malaysia. The disease is always associated with Moko disease caused by Ralstonia solanacearum. The bacterium forms irregular round, creamy colonies with red centres when cultured on Kelman’s tetrazolium chloride (TZC) medium. Moko disease has been interchangeably identified as blood disease as both of them cause similar wilting symptoms on banana. Therefore, the present study was carried out to detect the occurrence of bacterial wilt in Selangor through comprehensive procedures on morphology, molecular, pathogenicity and specificity tests. Isolation of bacteria was conducted on infected plant samples (fruit stalk, fruit pulps, stem and root) collected in the field by plating on TZC medium. Bacteria colonies that are Gram-negative, oxidase-positive, and obligate aerobe were chosen for further identification through polymerase chain reaction (PCR) using 759F and 760R primers. Molecular identification showed the bacteria that colonized the infected plants is confirmed as BDB, which is very closely related to R. solanacearum. After Koch’s Postulate and pathogenicity test on tomato, wilting symptoms were found on all banana plantlets after two weeks. The bacterium did not cause any wilting symptom on tomato plantlets. The present study revealed the distribution of blood disease in Selangor, after the first report in Perak. It also suggested that both morphology and DNA-based identification are necessary for the detection of the pathogen that causes bacterial wilt in banana. © 2016 Friends Science Publishers

Women in Katsina State, Nigeria have been using medicinal plants to cure various ailments associated with maternal health since time immemorial; however the use of such plants was never documented. In this study an ethnobotanical survey was conducted to document medicinal plants used for traditional maternal healthcare in Katsina State, Nigeria. Semi structured questionnaire method was used to interview 300 respondents (50 from 2 Local Government Areas of each of the 3 Senatorial Districts) comprising of herbalists, Traditional Birth Attendants (TBAs), Traditional Medical Practitioners (TMPs), House Wives, Farmers and others. Medicinal plants belonging to 101 genera distributed among 50 families were documented. Most of the reported plants belong to the Fabaceae (22.52%), Asteraceae (7.21%) Malvaceae (5.41%) and Anacardiaceae (4.51%) families. Acacia nilotica (L) Delile and Gueira senegalensis J.F. Gmel had the highest Relative Frequency of Citation (RFC) and Fidelity Level (FL) of 0.93; 100% and 0.92; 100%, respectively. Among the 18 categories of ailments, headache, navel pain, postpartum hemorrhage, and postpartum wound healing had the highest Informant Consensus Factor (ICF) of 1.00 each. Most of the reported plants (68.47%) were herbs and shrubs and about 84.68% of the surveyed plants were wild. Leaves were the most frequently used (32.14%) plant's part. Most of the medications (32%) were prepared as decoctions and preparations are mostly administered orally (84.68%). Scientific validation of the biological properties of the surveyed plants is highly advocated and cultivation of medicinal plants to minimize the pressure on wild species is also recommended.

Dendrobium mizanii, a new species to science belonging to Dendrobium Sect. Calcarifera was discovered in a summit region of a disturbed montane forest in Setiu, Terengganu, and named after His Majesty Sultan of Terengganu, Tuanku Mizan Zainal Abidin. A description, illustration, field and comparison with the closely related species D. crocatum from Peninsular Malaysia and D. doloissumbinii from Borneo are provided here.

Tributyltin (TBT) is a species of organotin compound (OTC), used as antifouling biocide in boat and ship paints to prevents the attachment of marine organism on their hull surfaces. Tributyltin was found to be very toxic to a variety of targeted and non-targeted organisms and has high persistence in sediments even after the total global ban by the International Maritime Organization (IMO) in 2008. Therefore, there is an urgent need to clean up TBT-polluted environments after the global banning due to the significant risks it poses to the human and aquatic organisms for its slow degradation rate. In selecting bioremediation agents, indigenous bacteria were documented to be of great potentials compared to non-indigenous. In this study, comparison was made between a bacterial isolate Klebsiella sp. FIRD 2, isolated from TBT-contaminated surface sediment and Pseudomonas specie isolated from non-TBT-contaminated soil. Previously, we isolated, screened, and identified Klebsiella sp. FIRD 2 as a TBT-resistant bacterium from TBT-contaminated surface sediment of Kong Kong Laut, Johor, Malaysia. The isolate was able to resist TBT up to 1500 μg/L without addition of carbon source in minimal salt medium (MSM). Pseudomonas sp., isolated from non-TBT-contaminated soil was tested in MSM treated with different concentration of TBT. The bacterium did not endure to survive in TBT-treated media without addition of carbon source; thus, the strain has no ability to utilize TBT as source carbon. Growth of Pseudomonas sp. was observed in MSM treated with TBT at concentration of 500 μg/L and 1000 μg/L along with addition of glucose as carbon source. No growth of Pseudomonas sp. was observed in MSM with higher TBT concentration even with additional of carbon source. This study equally endorses the potentials of indigenous bacteria in bioremediation of TBT contamination.

Vermicomposting is proven to be an effective way for nutrient cycling, converting large quantities of organic waste into value added product (vermicast). In order to harness the potential of earthworms in vermicomposting, selection of earthworm species that are able to consume large quantity of waste, and moreover, produce vermicast with high nutrient content is important. This experiment was carried out to compare the efficacy of Perionyx excavatus (PE) and Eudrilus eugeniae (EE) in vermicomposting rice straws. Ten earthworms were introduced into each vermibin containing grinded rice straw. The vermicast produced was collected periodically. The experiment was terminated when 70% of rice straw had decomposed. The plant nutrient contents and humic acids in vermicast were analysed. Vermicast of PE contained higher concentrations of total and available N, P, K and Mg while EE vermicast contained higher total and available Ca. Humic acid content was also found to be higher in EE vermicast. EE took 134 days while PE took 171 days to complete vermicomposting, thus, plant nutrient content generated per day in vermicast EE was higher compared to PE. Using EE in vermicomposting would contribute significantly towards practicing sustainable agricultural by recycling large amount of organic waste rice straw into value added high plant nutrient content vermicast. Key words: Earthworm biomass, decomposition rate, humic acid, agronomy.

The palm oil industry generates empty fruit bunch (EFB) in large quantities, which need to be optimally exploited to obtain maximum benefits from their uses. Vermicomposting of EFB with neem leaves is of great interest, as the end product might function as biofertiliser as well as biopesticide. Therefore, vermicomposting of different ratios of EFB and neem leaves using Eudrilus eugeniae as a composting agent was conducted. The results showed that the vermicompost produced was lower in pH, total organic carbon and the C/N ratio, but higher in humic acid content compared to the earthworm free control. The addition of neem leaves as a mixture of EFB-based vermicompost contributed to the enrichment in some important plant nutrients, such as total and available N, P, K and Ca. Results suggested that the addition of 10% neem leaves would enhance the earthworm biomass as well as nutrient contents, protease and phosphatase activity in vermicompost. Key words: Vermicomposting, empty fruit bunch, neem leaf, chemical properties, enzyme activity.

A study on fern family Gleicheniaceae was carried out for Peninsular Malaysia. A total of 14 species including 10 varieties in 4 genera was identified and recorded, genus Dicranopteris (4 species, 7 varieties), Diplopterygium (5 species), Gleichenia (3 species) dan Sticherus (3 species, 3 varieties). These ferns were collected from the fringes of the Lowland Dipterocarp Forest (LDF), Hill Dipterocarp Forest (HDF), Lower Montane Forest (LMF) and Upper Montane Forest (UMF). Four species and 3 varieties were found to be widely distributed over 3 types of vegetation, the LDF, HDF and LMF. Three species were distributed over higher elevation from the HDF, LMF to UMF. One variety, Dicranopteris linearis var. altissima was only recorded from the LDF, whilst Diplopterygium giganteum and Gleichenia dicarpa were recorded only in the UMF. Dicranopteris linearis and Sticherus truncatus were found to be the most widely distributed G. Rusea • L. Y. Chin • H. Omar • M. Muskhazli • Y. Umi Kalsom Biology Department, Faculty of Science, Universiti Putra Malaysia, 43400 UPM Serdang, Selangor, Malaysia Haja Maideen School of Environmental and Natural Resources Science, Faculty of Science and Technology, Universiti Kebangsaan Malaysia, 43600 UKM, Bangi, Selangor, Malaysia G. Rusea ( ) Biology Department, Faculty of Science, Universiti Putra Malaysia, 43400 UPM Serdang, Selangor, Malaysia e-mail: rusea@science.upm.edu.my species in this family whereby they could be found in all vegetation types. All species of Gleicheniaceae are terrestrial sun loving ferns and grew well on poor clay soils or poor nutrient soils. In fact, these ferns required small amount of mineral substances for growth. Members of this family mostly form thickets in open places, to which they are adapted to by their creeping rhizomes and the indefinite growth in length of scrambling or climbing fronds.

The chitinolytic system of Trichoderma sp. is complex with several types of chitinase working together simultaneously to enhance its mycoparasitism. However, chitinase enzyme has not been studied extensively for Trichoderma longibrachiatum although it has been proven to have significant effect as biocontrol. This study was conducted in the interest to purify chitinase 33kDA from T. longibrachiatum T28 and also to assess chit33 expression patterns. Chitinase activity was determined and the peak of chitinase activity was observed at 36 hours. Chitinase enzyme from the culture filtrate of T. longibrachiatum was precipitated with 80% acetone followed by anion- exchange chromatography using Neobar AQ. Only one chitinase band at 33kDa in size has been detected and analysis of Southern Blotting has confirmed the present of at least a single copy of chitinase 33kDA gene (chit33) for T. longibrachiatum strain T28. Assessment of chit33 expression showed that the expression was strongly affected by substrate specificity; where

A comprehensive study on the orchid diversity in Penang Hill, Penang, Malaysia was conducted from 2004 to 2008 with the objective to evaluate the presence of orchid species listed by Curtis (J Strait Br R Asiat Soc 25:67–173, 1894) after more than 100 years. A total of 85 species were identified during this study, of which 52 are epiphytic or lithophytic and 33 are terrestrial orchids. This study identified 57 species or 64.8% were the same as those recorded by Curtis (1894), and 78 species or 66.1% of Turner’s (Gardens’ Bull Singap 47(2):599–620, 1995) checklist of 118 species for the state of Penang including 18 species which were not recorded by Curtis (1894) and the current study but are actually collected from Penang Hill. A comparison table of the current findings against Curtis (1894) and Turner (1995) is provided which shows only 56 species were the same in all three studies. The preferred account for comparison was Curtis’ (1894) list as his report was specifically for the areas around Penang Island especially Penang Hill, Georgetown and Ayer Itam areas. Our study reveals that about 50% of Curtis’ collection localities have been converted to residential areas and agricultural land, and this probably explains the decreasing numbers of species found in the current study especially for the terrestrial species as epiphytic species have better adaptation capabilities towards environmental changes. Seven species were identified as new records to Penang Hill as they were not recorded by Curtis (1894). None of the three species recorded as endemic to Penang by Turner (1995) was recollected during the current study, of which only Zeuxine rupestris was in Curtis’ (1894) list. Overall, we concluded that Penang hill harbours at least 136 species of orchids of which 78 species or 57.4% were recollected in this study. This also indicates that this area is still suitable for orchid growth even though it is surrounded by rapid development and mass conversion of forested land into fruit orchards and residential area. The designation of Penang Hill as a Permanent Forest Reserve would better guarantee the survival of some orchid species unless human interventions and climatic changes occur.

Currently the development of green chemistry approach with the use of biomaterial-based activities of microbial cells in the synthesis of various nanostructures has attracted a great attention. In this study, we report on the use of bacterium, Bacillus cereus as a biotemplating agent for the formation of zinc oxide nanoparticles with raspberry- and plate-like structures through a simple thermal decomposition of zinc acetate by maintaining the original pH of the reaction mixtures. Possible mechanism on the formation of the nanostructures is proposed based on the surface chemistry and biochemistry processes involved organic-inorganic interactions between zinc oxide and the microbial cells.

Citinase 42 kDa produced by Trichoderma harzianum has been proven as a prime compund to be excreted onto the hyphae of the pathogen causing localised cell wall lysis at the point of interaction. This finally initiate the process of the host cell becomes empty of cytoplasm, disintegrates and shows a rapid collapse. This study investigates the existence of N-glyan linked mannose in chitinase 42 kDa from T.harzianum BIO10671 was initially purified using anion exchange chromatography prior to a series of experiments such as immunoblotting against the chitinase 42 kDa antibody, lectin staining for detecting any terminal linked mannose and galactofuranose detection to determine the presence of galactofuranose components in glycoproteins. The enzyme purification varvested about 12-fold of chitinase 42 kDa after overnight incubation in chitinase 42 kDa antibody suggesting taht the gene for chitinase 42 kDa was greatly expressed in this strain.There are no intervation of galactofuranose on any of the terminal mannose in chitinase 42 kDa as shown by negative results on samples treated with or without endoglycosidase-H and lectin staining. Therefore, it can be concluded that glycosylation occured in the chitinase 42 kDa from T.harianum 42 kDa was not in the form of N-glycan linked mannose as aspected.

Aims: Traditionally, crustacean wastes have been managed by using acid and alkali which leads to major environmental issue. However, over the recent years microbial fermentation has gained its way whereby producing similar effects as chemical treatment and a higher quality product can be obtained. Extracellular protease from Bacillus subtilis was used further by optimizing its culture medium to enhance protease production. Methodology and Results: The culture media was optimized with 4 various sources; Shrimp Crab Shell Powder (SCSP), nitrogen sources, inorganic salts, and carbon sources. It was found that culture media supplemented with 9% SCSP, 3% yeast extract, 1% sodium chloride and 9% glucose augmented protease activity up to 565.80 ± 19.41 U/mL compared to the un-optimized media (170.57 ± 6.75 U/mL). By using this optimized media, the ability and efficiency of B. subtilis in a period of 6 days was investigated whereby acid treated shrimp shells (ATSS) and raw shrimp shell powder (RSSP) were used in substitution of SCSP. In a period of 6 days, the protein content in both ATSS and RSSP was found to have been removed up to 60% and 42% respectively. However deproteinization was found to be more efficient in RSSP with the ratio of tyrosine to protein remained constantly high throughout the 6 days period. Conclusion, significance and impact of study: A better, more efficient and environmental friendly method is continuously being improvised to manage shrimp wastes with the use of microbes.

Earthworms are soil dwellers that have profound effects on soil ecosystem. Their feeding and burrowing activities help to incorporate soil particles with organic matter and redistribute them back into the soil via casts. The casts produced enhance microbial activities in soil that promote nutrient cycling. The present study was carried out to examine the physical, chemical and biological properties of casts produced by soil dwelling earthworm, Metaphire tschiliensis tschiliensis. Prior to inoculation of earthworms, the soil was incubated for two weeks at room temperature with moisture content of 20%. Worm worked soil and earthworm casts produced were collected for analysis after one month. Our results demonstrated that earthworm casts contained higher organic matter (9.84 ± 0.60%) and humic acid (1.33 ± 0.25%) compared to bulk soil. Total N, Ca, S, Al and Zn contents were also found to be higher in earthworm casts. Besides that, earthworm casts exhibited higher colony forming units (CFU), indicating the presence of higher microbial population. The study suggested that cast produced by M. tschiliensis tschiliensis could improve the physical and chemical properties of the soil. It also contained higher microbial population compared to soil without earthworm.