Method and quantity of liquid formulation of phosphobacteria required for seed inoculation

The survival of vegetative and sporulated cells of the Bacillus cultures on the seeds of the crop plants was tried in different combinations. One milliliter inoculum with 1 mL adhesive combination or sterile water showed better results followed by 1.5 mL inoculum with 0.5 mL adhesive or sterile water. The population of 5.5x10(5) cfu seed(-1) on black gram, 10.5x10(5) cfu seed(-1) on soybean and 6.5x10(5) cfu seed(-1) on maize were observed after 12 h of incubation in 1 mL sporulated inoculum mixed with 1 mL of rice gruel. The sporulated inoculum along with rice gruel favoured the adherence of the regenerated cells as rice gruel is rich in nutrient content.     

Microbiological quality of drinking water from dispensers in roadside restaurants of Bangladesh

The microbiological status of water from dispensers in different roadside restaurants of Dhaka city and Savar area was analyzed in this study. Seven samples from Dhaka and 8 samples of Savar were checked. The heterotrophic plate count was in a range of 1.0 x 10(3) CFU mL(-1) to 2.0 x 10(4) CFU mL(-1) (from new bottles), 1.0 x 10(3) to 1.5 x 10(4) CFU mL(-1) (after dispensation), and 1.5 x 10(3) CFU mL(-1) to 1.0 x l0(5) CFU mL(-1) (from serving glass). In several of the samples, the heterotrophic plate count was higher than the count in water from new bottle or after dispensation, suggesting added contamination from the serving glass. 80% of the samples were contaminated with total and fecal coliform bacteria, which render these waters unacceptable for human consumption. The samples were found to contain gram negative bacteria like E coli, Shigella sp., Klebsiella sp., Enterobacter sp., Pseudomonas sp., and Salmonella sp., which are potential pathogens and thus pose a serious threat to public health. This study elucidates the importance of monitoring the bottling companies and the restaurants and put them under strict regulations to prevent future outbreak of any water borne diseases caused by consumption of dispensed water.     

Antiviral activities of sulfated polysaccharides isolated from Sphaerococcus coronopifolius (Rhodophytha, Gigartinales) and Boergeseniella thuyoides (Rhodophyta, Ceramiales)

Water-soluble sulfated polysaccharides isolated from two red algae Sphaerococcus coronopifolius (Gigartinales, Sphaerococcaceae) and Boergeseniella thuyoides (Ceramiales, Rhodomelaceae) collected on the coast of Morocco inhibited in vitro replication of the Human Immunodeficiency Virus (HIV) at 12.5 μg/mL. In addition, polysaccharides were capable of inhibiting the in vitro replication of Herpes simplex virus type 1 (HSV-1) on Vero cells values of EC₅₀ of 4.1 and 17.2 μg/mL, respectively. The adsorption step of HSV-1 to the host cell seems to be the specific target for polysaccharide action. While for HIV-1, these results suggest a direct inhibitory effect on HIV-1 replication by controlling the appearance of the new generations of virus and potential virucidal effect. The polysaccharides from S. coronopifolius (PSC) and B. thuyoides (PBT) were composed of galactose, 3,6-anhydrogalactose, uronics acids, sulfate in ratios of 33.1, 11.0, 7.7 and 24.0% (w/w) and 25.4, 16.0, 3.2, 7.6% (w/w), respectively.     

Bioassay evaluation of the entomopathogenic fungi, Beauveria bassaina Vuellemin against eggs and nymphs of Bemisia tabaci Gennadius (Homoptera: Aleyrodidae)

This study was carried out to determine the lethal effect of the entomopathogenic fungi, Beauveria bassaina Vuell. on eggs, young and old nymphs of the whitefly, Bemisia tabaci Genn. Mortality percentage was significantly differed based on stage of B. tabaci and conidial concentrations of B. bassina. Average of the infection level to insect was very low particularly in eggs with only 4.49%, even with higher conidial concentrations (6 x 10(6) conidia mL(-1)). Whereas, it was higher with 1st and 2nd instars (42.045%) and 3rd and 4th instars (35.93%). Three parameters was assessed with B. tabaci eggs, namely; egg infection, egg hatchability and crawlers emergence. Egg mortality percentages averaged 1.2, 4.27 and 8.0% with fungal concentration 2 x 10(6), 4 x 10(6) and 6 x 10(6) conidia mL(-1), respectively. Daily infection percentages were varied depend upon the conidial concentration where the highest infection rate of eggs was occurred with 6 x 10(6), followed by 4 x 10(6) conidia mL(-1). Egg hatch was very high, while the mortality among the emerged crawlers was neglectable compared with the check. Efficiency of B. bassaina on whitefly nymphs also was varied based on the insect instar and fungal concentration. Mortality percentages were obviously higher to young nymphs (1st and 2nd instars) than to older ones (3rd and 4th instars). The results indicated that nymphs were highly susceptible to fungal treatment compared with eggs. Additionally, pathogenicity and virulence of B. bassaina against B. tabaci immatures was not indicated by LC50 only, but also, by the time in days (LT50) required to achieve 50% mortality of an insect.     

Cloning and expression of coxsakievirus B3 viral protein-1 in E. coli

BACKGROUND: Viral protein-1 (VP1) is a major capsid protein of Coxsakievirus B3 (CVB3) that plays an important role in directing viruses towards permissive cells and acts as a main antigenic site of the virus in eliciting of host immune response, hence it seems VP1 can be considered as a vaccine candidate against CVB3 infection. In this study, cDNA of VP1 was prepared, cloned into pET expression vector and the recombinant protein (VP1) was over expressed in E. coli. METHODS: The viruses were grown in suspension cultures of Vero cells with an input virus multiplicity of 10-50 plaque-forming units/cell. After observing complete cytopathic effect, the total RNA (cells and virus) was prepared for RT-PCR and by using specific primers, VP1 cDNA was amplified and ligated into pET vectors (32 a and 28 a). The recombinant vector was transferred into competent E. coli (BL-21) and after selection of proper colony, which carried correct cDNA within the vector; cells were cultured and induced with isopropyl B-D-thiogalactopyranoside, in order to express protein (VP1). The cultures were tested for presence of VP1 by SDS-PAGE and Western-Blotting analysis. RESULTS: Molecular techniques such as PCR which showed exact defined size of the VP1 (819 bp), restriction digestion and finally immunoblot analysis of over expressed protein; all confirmed the correct cloning and expression of VP1 in this research. CONCLUSION: In this research, full length of VP1 as major capsid protein of CVB3 was over expressed in E. coli which, can be used for further studies, including neutralizing antibody production against CVB3.     

Evaluation of different media and methods of cultivation on the production and viability of entomopathogenic fungi, Verticillium lecanii (Zimm.) Viegas

For mass production of V. lecanii, three types of cultivation methods including liquid, solid and diphasic production systems were investigated. In the liquid state of production, six media were tested in stationary culture conditions. Among the six media tested, Molasses Yeast Broth (MYB) supported maximum sporulation (8.33 x 10(8) spores mL(-1)) and biomass production (746 mg/100 mL). In the MYB, 4% molasses concentration was found to produce highest spore count (8.56 x 10(8) spores mL(-1)) and biomass (776 mg/100 mL) followed by 5 and 6% molasses. Among the six solid substrates tested, rice grains supported highest spore production (1.14 g/100 g). In diphasic state of production, combination of MYB and rice grains produced the greatest amount of spores, (1.70 g/100 g). Results of this study indicated that diphasic method using MYB as liquid medium and rice as solid substrate are the best method and media for mass production of V1-7 isolate of V. lecanii.     

Role of bacteria isolates in the spoilage of fermented African oil bean seed ugba

Study was carried out to determine the spoilage association of traditionally fermented ugba, product of African oil bean seed. Samples were collected from three markets within Enugu metropolis in Eastern Nigeria. Microbial population of ugba was isolated, characterized and identified. Effect of time on product quality, total viable cell count and individual growth pattern of isolates was evaluated. Microorganisms isolated were Proteus sp., Klebsiella sp., Staphylococcus epidermidis, E. coli, Bacillus licheniformis and Bacillus subtilis. Initial viable cell count increased from 4.6 x 10(8) -6.2 x 10(8), 6.0 x 10(9) -6.9 x 10(9) and 3.9 x 10(8) -5 x 10(8) cfu g(-1) by the 4th day for samples from Mayor, Kenyatta and Garki markets, respectively. Organoleptic changes in texture and colour were seen to be a factor of time of storage. A decline in growth of Proteus sp., E. coli, Klebsiella sp. and S. epidermidis by the 4th day of storage was observed for all samples. Viable cells of B. licheniformis increased from 6.1 x 10(7) -17.7 x 10(7) cfu g(-1), 13.9 x 10(8) -20.0 x 10(8) and 8.7 x 10(7) -15.5 x 10(7) by the 4th day of storage for samples from Mayor, Kenyatta and Garki markets respectively. Growth of B. subtilis increased from 9.2 x 10(7) -19.9 x 10(7), 14.9 x 10(8) -21.2 x 10(8) and 11.5 x 10(7) -17.2 x 10(7) cfu g(-1) for samples from Mayor, Kenyatta and Garki markets respectively. Our results indicate that ugba spoilage is primarily a result of the continued activity of African oil bean seed fermentative organisms B. subtilis and B. licheniformis.     

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.     

Diploid and tetraploidSolanum chacoense genotypes that synthesize leptine glycoalkaloids and deter feeding by Colorado potato beetle

A selection (8380-1) fromSolatium chacoense Bitter (2n=2x=24) accession PI 458310 that synthesizes the leptine glycoalkaloids was compared in growth chambers with tetraploid (2n=4x=48) genotypes derived from tissue culture of 8380-1 leaf expiants for plant growth habit, leaf glycoalkaloid content, and effect on the development of Colorado potato beetle [Leptinotarsa decemlineata (Say)] larvae. The plants of the 4x regenerant genotypes were more vigorous with larger, more oval shaped leaflets than 8380-1 plants. The leaf concentrations of leptines and total glycoalkaloids were significantly lower (about 34%) in the 4x genotypes than in 8380-1. The proportion of leptines in the total glycoalkaloid content was nearly the same (about 80%) in both ploidy groups. In leaf-disk feeding tests, the development of Colorado potato beetle neonate larvae was not significantly different for the 2x and 4x genotypes. Both groups significantly slowed development compared with development on cv. Kennebec leaf disks. The 8380-1 selection and a group of 4x 8380-1 regenerant genotypes are maintained in the Vegetable Laboratory, Beltsville Agricultural Research Center, Beltsville, MD 20705 and are available for distribution.     

Influence of the Structural Features of Carrageenans from Red Algae of the Far Eastern Seas on Their Antiviral Properties

The structural diversity and unique physicochemical properties of sulphated polysaccharides of red algae carrageenans (CRGs), to a great extent, determine the wide range of their antiviral properties. This work aimed to compare the antiviral activities of different structural types of CRGs: against herpes simplex virus type 1 (HSV-1) and enterovirus (ECHO-1). We found that CRGs significantly increased the resistance of Vero cells to virus infection (preventive effect), directly affected virus particles (virucidal effect), inhibited the attachment and penetration of virus to cells, and were more effective against HSV-1. CRG1 showed the highest virucidal effect on HSV-1 particles with a selective index (SI) of 100. CRG2 exhibited the highest antiviral activity by inhibiting HSV-1 and ECHO-1 plaque formation, with a SI of 110 and 59, respectively, when it was added before virus infection. CRG2 also significantly reduced the attachment of HSV-1 and ECHO-1 to cells compared to other CRGs. It was shown by molecular docking that tetrasaccharides—CRGs are able to bind with the HSV-1 surface glycoprotein, gD, to prevent virus–cell interactions. The revealed differences in the effect of CRGs on different stages of the lifecycle of the viruses are apparently related to the structural features of the investigated compounds.     

3-D Culture of Marine Sponge Cells for Production of Bioactive Compounds

Production of sponge-derived bioactive compounds in vitro has been proposed as an alternative to wild harvest, aquaculture, and chemical synthesis to meet the demands of clinical drug development and manufacture. Until recently, this was not possible because there were no marine invertebrate cell lines. Recent breakthroughs in the development of sponge cell lines and rapid cell division in improved nutrient media now make this approach a viable option. We hypothesized that three-dimensional (3-D) cell cultures would better represent how sponges function in nature, including the production of bioactive compounds. We successfully cultured sponge cells in 3-D matrices using FibraCel^® disks, thin hydrogel layers, and gel microdroplets (GMDs). For in vitro production of bioactive compounds, the use of GMDs is recommended. Nutrients and sponge products rapidly diffuse into and out of the 3-D matrix, the GMDs may be scaled up in spinner flasks, and cells and/or secreted products can be easily recovered. Research on scale-up and production is in progress in our laboratory.     

Sulfated Polysaccharides from Seaweed Strandings as Renewable Source for Potential Antivirals against Herpes simplex Virus 1

Herpes simplex virus 1 (HSV-1) remains a prominent health concern widespread all over the world. The increasing genital infections by HSV-1 that might facilitate acquisition and transmission of HIV-1, the cumulative evidence that HSV-1 promotes neurodegenerative disorders, and the emergence of drug resistance signify the need for new antiviral agents. In this study, the in vitro anti-herpetic activity of sulfated polysaccharides (SPs) extracted by enzyme or hot water from seaweeds collected in France and Mexico from stranding events, were evaluated. The anti-herpetic activity evaluation of the semi-refined-polysaccharides (sr-SPs) and different ion exchange purified fractions showed a wide range of antiviral activity. Among them, the sr-SPs from the Rhodophyta Halymenia floresii showed stronger activity EC₅₀ 0.68 μg/mL with SI 1470, without cytotoxicity. Further, the antiviral activity of the sr-SPs evaluated at different treatment schemes showed a high EC₅₀ of 0.38 μg/mL during the viral adsorption assays when the polysaccharide and the virus were added simultaneously, whilst the protection on Vero cell during the post-infection assay was effective up to 1 h. The chemical composition, FTIR and ¹H NMR spectroscopic, and molecular weights of the sr-SPs from H. floresii were determined and discussed based on the anti-herpetic activity. The potential utilization of seaweed stranding as a source of antiviral compounds is addressed.     

Anti HSV-1 Activity of Halistanol Sulfate and Halistanol Sulfate C Isolated from Brazilian Marine Sponge Petromica citrina (Demospongiae)

The n-butanol fraction (BF) obtained from the crude extract of the marine sponge Petromica citrina, the halistanol-enriched fraction (TSH fraction), and the isolated compounds halistanol sulfate (1) and halistanol sulfate C (2), were evaluated for their inhibitory effects on the replication of the Herpes Simplex Virus type 1 (HSV-1, KOS strain) by the viral plaque number reduction assay. The TSH fraction was the most effective against HSV-1 replication (SI = 15.33), whereas compounds 1 (SI = 2.46) and 2 (SI = 1.95) were less active. The most active fraction and these compounds were also assayed to determine the viral multiplication step(s) upon which they act as well as their potential synergistic effects. The anti-HSV-1 activity detected was mediated by the inhibition of virus attachment and by the penetration into Vero cells, the virucidal effect on virus particles, and by the impairment in levels of ICP27 and gD proteins of HSV-1. In summary, these results suggest that the anti-HSV-1 activity of TSH fraction detected is possibly related to the synergic effects of compounds 1 and 2.     

Effects of different concentrations of bovine follicular fluid and estrous cow serum on development of murine 2-cell embryos

Murine 2-cells embryos were isolated from murine oviducts at laboratory and transferred into Ham's F-10 medium containing 0.1 mg mL(-1) streptomycin and 100 IU mL(-1) penicillin G and supplemented with 3 mg mL(-1) bovine serum albumin (BSA) or different concentrations of bovine follicular fluid (bFF) and estrous cow serum (ECS). Significantly higher (p<0.05) > or =4-cell embryos were developed when embryos were cultured 20% bFF (84.33%) comparing to 10 and 15% bFF (48.33 and 69.33%) as well as 3 mg mL(-1) BSA (65.66%). Morula rates were also lower in 10% bFF (22.33%) comparing to the other groups and were similar in 15 and 20% bFF (62.66 and 72.33% morula rates) as well as BSA containing media (55.33%). The highest (p<0.05) blastocyst rates were obtained in medium containing 20% bFF (64.33%) and the lowest belonged to 10% bFF (15%) comparing to 15% bFF (33.66%) or 3 mg mL(-1) BSA. When embryos were cultured in ECS, no significant different was observed in different culture media (76.66, 72.33, 82.5 and 65.66% > or =4-cell embryos in 10, 15 and 20% bFF and 3 mg mL(-1) BSA, respectively). Morula and blastocyst rates were also similar in all groups (32.33, 41.66 and 66.25 and 55.33% morula rates and 15.33, 27, 44.50 and 29.66% blastocyst rates for 10, 15 and 20% bFF and 3 mg mL(-1) BSA, respectively). The results of the present study demonstrated that 20% bFF could be substituted for BSA when in vitro culture of murine embryos is carried.     

大豆菌核病流行预测研究

１９８８－１９９２年，在黑龙江省北部研究了与大豆菌核病发病率显著相关的因子。在菌核史较重的田块中，大豆菌核病的发病率和大豆开花期间的气象因子，表现了显著的相关。经电子计算机进行多元逐步回归分析，建立了大豆菌核病年发病程度拟合方程：Ｙ＝－１２８．３２８＋０．１１５５１ｘ１＋４．０６０６４ｘ２－０．１２５６９ｘ３，复相关系数Ｒ＝０．９９９９９３，经检验，理论值和实际调查值拟合很好。通过大豆盛花期间田间     

近地表臭氧浓度升高对不同东北品种大豆产量形成及品质的影响

为明确大气臭氧浓度升高对不同东北大豆品种产量形成和品质的影响,本研究以环境大气臭氧浓度为对照,利用开顶式气室模拟大气臭氧浓度升高40 nL·L^-1,选取3个推广面积较大的大豆品种绥农4号、绥农8号和东生1号,研究在高臭氧浓度下不同大豆品种产量形成和品质的差异。结果表明:臭氧浓度升高对大豆产量形成具有明显的负作用,3个大豆品种的产量平均降低33%。不同大豆品种产量对臭氧浓度升高的响应存在明显差异,产量降低幅度:绥农4号(-41%)>东生1号(-36%)>绥农8号(-23%)。利用逐步回归法分析发现大气臭氧浓度升高条件下大豆籽粒大小(x_1)和单株总荚数(x_2)变化可以用于估计大豆产量变化,两者与大豆产量的回归方程:y=-3.639+0.098x_1-0.285x_2。此外,大气臭氧浓度升高对大豆品质产生明显影响,3个大豆品种籽粒中蛋白质浓度均显著提高2.8%～4.6%(P<0.05),而脂肪含量则显著降低4.2%～7.8%(P<0.05)。     

氧化应激对Vero细胞的损伤及EGCG的保护作用

研究了氧化应激对肾细胞Vero细胞的损伤及EGCG的保护作用。用MTT、吖啶橙染色和DNA凝胶电泳等方法研究了H2O2和Cr6+应激对Vero细胞的损伤,及EGCG对凋亡细胞的保护作用及其机理。结果表明,H2O2和Cr6+剂量效应地抑制Vero细胞活性,IC50分别为175.6和9.8mgL-1;其中50 mgL-1 H2O2和400 mgL-1/2h Cr6+可诱导Vero细胞凋亡。0~60 mgL-1 EGCG有效抑制H2O2和Cr6+应激引起的细胞活性下降,且40 mgL-1 EGCG显著抑制细胞凋亡。对于Cr6+所诱导的细胞凋亡,EGCG的保护作用与EDTA和Vc的协同作用效果相当,表明清除活性氧和络合金属离子都有助于减轻Cr6+应激损伤。可见,EGCG通过清除活性氧和络合离子有效地保护了肾细胞免受应激损伤,这对易遭受活性氧损伤的肾脏无疑具有一定的临床价值。     

Antiviral Potential of Sea Urchin Aminated Spinochromes against Herpes Simplex Virus Type 1

Herpes simplex virus type 1 (HSV-1) is one of the most prevalent pathogens worldwide requiring the search for new candidates for the creation of antiherpetic drugs. The ability of sea urchin spinochromes—echinochrome A (EchA) and its aminated analogues, echinamines A (EamA) and B (EamB)—to inhibit different stages of HSV-1 infection in Vero cells and to reduce the virus-induced production of reactive oxygen species (ROS) was studied. We found that spinochromes exhibited maximum antiviral activity when HSV-1 was pretreated with these compounds, which indicated the direct effect of spinochromes on HSV-1 particles. EamB and EamA both showed the highest virucidal activity by inhibiting the HSV-1 plaque formation, with a selectivity index (SI) of 80.6 and 50.3, respectively, and a reduction in HSV-1 attachment to cells (SI of 8.5 and 5.8, respectively). EamA and EamB considerably suppressed the early induction of ROS due to the virus infection. The ability of the tested compounds to directly bind to the surface glycoprotein, gD, of HSV-1 was established in silico. The dock score of EchA, EamA, and EamB was −4.75, −5.09, and −5.19 kcal/mol, respectively, which correlated with the SI of the virucidal action of these compounds and explained their ability to suppress the attachment and penetration of the virus into the cells.     

Buthionine sulfoximine inhibits cytopathic effects and apoptosis induced by infection with AIK-HDC strain of measles virus

BACKGROUND: Measles virus (MV) is a highly contagious agent which causes a major health problem in developing countries. We studied the effect of buthionine sulfoximine (BSO) on the replication of an AIK-HDC strain of MV and its induced apoptosis in Vero cell lines. METHODS: In this study, toxicity of BSO on Vero cells was investigated first, resulted in determination of sub-lethal or non-toxic concentration zone of BSO for cells. Next, anti-viral effect of BSO at various time limits was evaluated and virus titer was determined at each stage either as 50% tissue culture infective dose (TCID) 50 or by plaque assay method. Using specific anti-measles IgG, anti-viral effect of BSO on MV replication cycle was evaluated through indirect immunofluorescence assay, meanwhile presence of viral RNA was investigated by RT-PCR and gel electrophoresis. RESULTS: According to the experiments, BSO, at concentration of 50 microM, markedly inhibited the cytopathic effect (CPE) induced by MV. BSO also significantly inhibited apoptosis induced by MV. BSO either influences replication of MV genome, or may inhibit virion formation. CONCLUSION: These results suggest that the inhibition of CPE and apoptosis by BSO induced by MV may be associated with the effect of BSO on viral RNA genome. Therefore, it is suggested that MV infections can induce apoptosis through the activation of a common pathway that can be inhibited by BSO.     

Internalisation of microbes in vegetables: microbial load of Ghanaian vegetables and the relationship with different water sources of irrigation

The occurrence of pathogens in the internal parts of vegetables is usually associated with irrigation water or contaminated soil and could pose risk to consumers as the internalised pathogens are unaffected by external washing. This study was carried out to assess the rate of internalisation of microbes in common Ghanaian vegetables. Standard microbiological methods were employed in microbial enumeration of vegetables collected at the market and farm levels, as well as irrigation water and soil samples. The overall mean counts of vegetables were 4.0 x 10(3) cfu g(-1); 8.1 x 10(2) cfu g(-1); 2.0 x 10(2) cfu g(-1); 3.5 x 10(2) cfu g(-1) for total bacteria, coliform counts, faecal coliform counts and yeast counts, respectively. The rate of internalisation of coliforms in vegetables irrigated with stream/well water was 2.7 times higher than those irrigated with pipe water. The mean coliform counts (4.7 x 10(7) cfu g(-1)) and faecal coliform counts (1.8 x 10(6) cfu g(-1)) of soil samples were similar to those of stream water suggesting both sources exerted similar contamination rates on the vegetables. Generally, there were no significant variations between the rates of internalisation of microbes at the market and farm levels at p < 05, indicating that internalisation of microbes in the vegetables mainly occurred at the farm level. The study has shown that microbial contamination of vegetables in Ghana is not limited to the external surface, but internal vegetable parts could harbour high microbial loads and pose risk to consumers. Safety practices associated with the commodity should therefore not be limited to external washing only. There is the additional need of heating vegetables to eliminate microbes both externally and internally before consumption.