X107 CFU Research Articles

Sacubitril/valsartan alleviates sepsis-induced myocardial injury in rats via dual angiotensin receptor-neprilysin inhibition and modulation of inflammasome/caspase 1/IL1β pathway

Sepsis is a life-threatening situation that ultimately affects cardiac function, leading to cardiomyopathy and myocardial injury as a result of uncontrolled response to infection.Till now, there is limited effective treatment to rescue those cases. Thus, novel therapeutic strategies should be identified to achieve better outcomes for septic patients. For the first time, we aimed to evaluate the effect of sacubitril/valsartan (Sac/Val) on sepsis-induced cardiac injury. Wistar male adult albino rats were randomly divided into four groups; Group I received the vehicle; Group II was given the vehicle plus 1 ml saline containing viable Escherichia coli (E. coli) (2.1 × 109 cfu) by intraperitoneal (i.p.) injection on the 1st and 2nd days; Group III received i.p. injection as group II plus oral administration of Sac/Val (30 mg/kg/day) and Nitro- ω-L-arginine (L-NNA) (25 mg/kg/day) for 7 days. Group IV was administered i.p. injection as group II plus oral administration of Sac/Val (30 mg/kg/day) for 7 days. Our data (n = 10) revealed successful induction of sepsis as it showed a significant increase in the measured cardiac enzymes, malondialdehyde (MDA), angiotensin II (Ang II), neprilysin, inflammasome, caspase 1, interleukin (IL)1β, and caspase 3 with cardiac histopathological changes, but there was a significant decrease in the antioxidants and blood pressure (BP). Co-administration of Sac/Val could obviously improve these changes. Interestingly, L-NNA given group showed a decrease in the cardioprotective effect of Sac/Val. Sac/Val could ameliorate sepsis induced cardiac damage via inhibition of Ang II and neprilysin with anti-inflammatory, anti-oxidant and anti-apoptotic properties.

Microbial load and aflatoxin content of street foods sold around Federal University of Technology, Akure (FUTA) campus

The microbial density and diversity, as well as the presence of aflatoxin in selected street vended-foods sold in FUTA campus was investigated in this study. Seven selected food samples viz; cooked rice, raw rice, garri, eba, peanut, suya and pap, were purchased from vendors at four popular locations around the school campus. The microbial load and diversity in the foods were analyzed using standard methods. The bacterial counts ranged from 3.1 x 104 CFU/ml to 4.9 x105 CFU /ml, with peanut sample having the highest count and cooked rice having the lowest count. Bacterial isolates belonging to eleven genera (Staphylococcus, Streptococcus, Campylobacter, Listeria, Bacillus, Clostridium, Salmonella, Klebsiella, Pseudomonas, Enterobacter, Escherichia) were obtained from the food samples. Mean Fungal counts of food samples ranged from 3.2 x 104 to 1.29 x106 SFU/ml. The fungi isolated include species of Aspergillus, Penicillium, Fusarium, Cladosporium, Cryptococcus and Candida. The high count of microorganisms coupled with the presence of Salmonella typhimurium and aflatoxins in some of the food samples portend danger to the consumers of such foods.

Bio-Prospecting Xylose-Utilizing, Exopolysaccharide (EPS)-Producing Bacteria and EPS Quantification through Submerged Fermentation using Xylose as the Major Carbon Source

Study’s Novelty/ Excerpt This study identified bacterial isolates from diverse environments capable of utilizing xylose for exo-polysaccharide (EPS) production, an area with limited prior research. Potential EPS producers were isolated and identified as Staphylococcus, Enterobacter, Kocuria, Klebsiella, Enterococcus, Serratia, and Burkholderia species using a salt-based medium with xylose as the primary carbon source. The highest EPS production was observed in Enterobacter cloacae D1, E. cloacae D2, Klebsiella oxytoca D2, and K. oxytoca G1, ranging from 0.04 g/L to 2.0 g/L, highlighting these isolates' potential for optimization and application in various industries. Full Abstract Many microorganisms are capable of producing Exo-polysaccharides (EPS) while utilizing simple sugars and hexoses. These EPS found applications in various fields, such as agricultural biotechnology, pharmaceuticals, textiles, and food industries. However, there is a lack of studies on EPS-elaborating bacteria that can utilize pentoses like xylose. Therefore, the utilization of alternative carbon sources for EPS production has become a focus of recent research. This study aimed to prospect bacteria that can utilize xylose for EPS production. Samples from agricultural soil, dump sites, saline soil, cement-contaminated soil, fresh cow milk, cow dung, and yogurt were serially diluted and cultured in a salt-based medium with xylose as the primary carbon source. Slimy and mucoid colonies were selected as potential EPS-producing isolates and identified morphologically and biochemically using the VITEK 2 Automated identification system. The quantification of EPS production by these isolates was conducted through submerged fermentation with xylose as the sole carbon source. The mean heterotrophic bacterial count of xylose-utilizing bacteria ranged from 2.1x106 CFU to 3.5x108 CFU per gram of the samples analyzed. The slimy and mucoid colonies were identified as members of the genera Staphylococcus, Enterobacter, Kocuria, Klebsiella, Enterococcus, Serratia, and Burkholderia. The quantities of EPS produced by the isolates ranged from 0.04 g/L to 2.0 g/L, with E. cloacae D1, E. cloacae D2, K. oxytoca D2, and K. oxytoca G1 elaborating the highest amount of EPS. Bacterial isolates capable of utilizing xylose for EPS production were obtained from various sources, showing potential for further optimization

A Rapid and Robust DNA Extraction Method for PCR-Based Diagnosis of V. cholerae

Cholera, caused by _V. cholerae_, needs rapid diagnosis because of its threat for rapid spread. Recent molecular diagnosis by PCR assay is more advantageous than the conventional methods. The bottleneck in PCR diagnosis lies in the delay in template DNA extraction of _V. cholerae_, which is the basic requirement. This study describes a simple, less expensive, and rapid template DNA extraction method to lessen the total time for PCR diagnosis of _V. cholerae._ To obtain template DNA, our method involves boiling of _V. cholerae_ suspension in distilled water (takes 18-24 hours) instead of boiling the suspension in broth medium, which is a lengthy process (takes 72 hours). The validation of our template DNA was conducted using 40 _V. cholerae_ O1 strains that were confirmed previously, and its usefulness was checked on 20 clinical strains of _V. cholerae_ O1 isolated from acute diarrhea patients; results were compared with those of conventional template DNA. The analysis of our template DNA in simplex and quadruplex PCR assays revealed the presence of _ctxA_, _tcpA _(El Tor), _rfb _(O1), _ToxR_, _ctxB_, _zot_, _ace,rst _, and _OmpW _genes in _V. cholerae _O1. The comparison of the results of PCR assays using the template DNA from both sources showed equal results and matched correctly. The sensitivity of our template DNA was 1.5x103 CFU per assay. Our template DNA extraction method is simple, less expensive, and rapid, and it can be employed for early diagnosis of _V. cholerae_ during outbreaks, in research laboratories, and in hospital setups where infrastructures are available.

RHIZOSPHERE MICROBIOME, SOIL ENZYMES AND GROUNDNUT YIELD AS INFLUENCED BY ADDITION OF BIOCHAR AND FERTILIZERS IN IRRIGATED ALFISOL OF NORTH COASTAL ANDHRA PRADESH

The field experiment was conducted in irrigated Alfisols of North coastal Andhra Pradesh tostudy the effect of biochar and chemical fertilizers on rhizosphere microbiome, soil enzymes andyield performance of groundnut crop (variety K-6) during rabi, 2018-19. Biochar application tosoil significantly increased soil bacterial, fungal and actinomycetes population. At pod developmentstage, the highest number of bacterial count (39.0 x106 CFU g-1 soil) was observed in T5 (100%RDF + biochar @ 6 t ha-1) which was on par with all the biochar applied treatments (T3, T4, T6, T7,T8). The Biochar applied @ 6 t ha-1 (T5 & T8) significantly increased the bacterial population ascompared to non-biochar applied treatments (T1 & T2). Fungal and actinomycetes populationfollowed the similar trend of bacterial count. Soil urease activity was significantly superior inbiochar applied treatments (T3, T4, T5, T6, T7, T8) compared to non biochar applied treatments (T1and T2). With increased rates of biochar application the urease activity markedly increased insoil. Similar trend was noticed with respect to dehydrogenase, acid phosphatase and alkalinephosphatase enzymes in soil in response to addition of biochar. At pod development stage, thehighest leaf area index (3.16) was recorded with 100% RDF + biochar @ 6 t ha-1(T5) which wassignificantly higher than T1 (control), T2 (100% RDF) and T6 (75% RDF + biochar @ 2 t ha-1). Thehighest drymatter accumulation of 2951 kg ha-1 at peg penetration and 6428 kg ha-1 at poddevelopment was observed in T5 (100% RDF + biochar @ 6 t ha-1) which was on par with T3(100% RDF + biochar @ 2 t ha-1), T4 (100% RDF + biochar @ 4 t ha-1), T8 (75% RDF + biochar @6 t ha-1) treatments. Groundnut pod yield was highest (4019.58 kg ha-1) in treatment received100% RDF + biochar @ 6 t ha-1, which was on par with T4 (100% RDF + biochar @ 6 t ha-1) and T8(75% RDF + biochar @ 6 t ha-1).

Enhancing iron bioavailability in lentil grains circumventing the phytate barrier

The study aimed at examining the effect of Iron (Fe) management strategies on the redistribution of Fe fractions in soil and on the Fe bioavailability and phytic acid (PA) content in lentil grains. A pot experiment was conducted in Completely Randomized Design with 8 treatments replicated thrice. The treatments were Control (no fertilizer); Recommended dose of fertilizers (RDF) i.e. 20:40 (N: P2O5); RDF + Fe (basal application @10 kg/ha); RDF + Fe (foliar@ 1% (w/v) solution); RDF + Fe (basal + foliar); RDF + FeSB (biopriming with Fe solubilizing bacteria (Enterobacter cloacae strain BAU3) @ 5 mL/kg seed (1 x109 cfu mL−1)); RDF + Fe (seed nutripriming with 1% (w/v) solution); and RDF + (biopriming + nutripriming). Fe source for the treatments was Fe-EDDHA chelate. Higher Fe release from these fractions was observed under biopriming + nutripriming with water soluble Fe of 21.09 and 20.82 mg kg−1, respectively for the biopriming + nutripriming and sole biopriming treatments; and organically bound Fe of 230.95 and 211.36 mg kg−1 for the same treatments. These fractions contributed toward higher Fe uptake which was highest under biopriming + nutripriming. Values were statistically significant for biopriming and nutripriming and basal + foliar treatment. Lowest PA values in lentil grains as well as PA/Fe ratio were estimated under basal + foliar and biopriming + nutripriming treatments. Combined basal and foliar application of Fe esp. in chelated form and use of seed priming strategies have immense potential in enhancing the Fe bioavailability in grains from soil, seed or foliage.

Defining a metagenomic threshold for detecting low abundances of Providencia alcalifaciens in canine faecal samples.

Acute haemorrhagic diarrhoea syndrome (AHDS) in dogs is a condition of unknown aetiology. Providencia alcalifaciens is suspected to play a role in the disease as it was commonly found in dogs suffering from AHDS during a Norwegian outbreak in 2019. The role of this bacterium as a constituent of the canine gut microbiota is unknown, hence this study set out to investigate its occurrence in healthy dogs using metagenomics. To decrease the likelihood of false detection, we established a metagenomic threshold for P. alcalifaciens by spiking culture-negative stool samples with a range of bacterial dilutions and analysing these by qPCR and shotgun metagenomics. The detection limit for P. alcalifaciens was determined and used to establish a metagenomic threshold. The threshold was validated on naturally contaminated faecal samples with known cultivation status for P. alcalifaciens. Finally, the metagenomic threshold was used to determine the occurrence of P. alcalifaciens in shotgun metagenomic datasets from canine faecal samples (n=362) collected in the HUNT One Health project. The metagenomic assay and qPCR had a detection limit of 1.1x103 CFU P. alcalifaciens per faecal sample, which corresponded to a Cq value of 31.4 and 569 unique k-mer counts by shotgun metagenomics. Applying this metagenomic threshold to 362 faecal metagenomic datasets from healthy dogs, P. alcalifaciens was found in only 1.1% (95% CI [0.0, 6.8]) of the samples, and then in low relative abundances (median: 0.04%; range: 0.00 to 0.81%). The sensitivity of the qPCR and shotgun metagenomics assay was low, as only 40% of culture-positive samples were also positive by qPCR and metagenomics. Using our detection limit, the occurrence of P. alcalifaciens in faecal samples from healthy dogs was low. Given the low sensitivity of the metagenomic assay, these results do not rule out a significantly higher occurrence of this bacterium at a lower abundance.

The influence of plant components on microbiological and oxidative processes in dry fish and plant products during storage

A pressing issue in the food industry is providing the population with adequate nutrition. According to the concept of healthy eating, a person needs to increase the consumption of plant foods without reducing the consumption of foods containing complete protein. We have created dry fish and plant products, in the form of food concentrates, containing all the necessary macro- and micronutrients. The purpose of the research was to study the influence of plant components on the microbiological and oxidative processes of dry fish and fish-plant products during storage. To achieve the intended goal, the tasks of determining microbiological indicators and oxidation products at the beginning and end of the storage period were solved. Storage of dry fish and plant products was carried out for 12 months in polymer packaging at a temperature of plus 20±5℃. All analyses were carried out using generally accepted standard methods. As a result of the studies, it was established that the microbiological indicators of a dry fish-plant product, due to the addition of vegetables and corn flour, were higher than those of a dry fish product (0.5x104 and 0.2x104 CFU per 1 g, respectively), but during storage the bacterial contamination of dry of fish and plant products decreased from 0.5x104 to 0.1x104 CFU per 1 g of product. Also, plant components affected the oxidative processes in dry fish and vegetable products. The acid number in samples of dry fish product increased from 0.71 to 2.67 mg of KOH, and in samples of dry fish-plant product – from 0.65 to 1.87 mg of KOH after 12 months of storage. Thus, plant components inhibited the oxidation process and maintain the quality of the product during storage.

Effect of Agro-Waste on Bioremediation of Soil Polluted with Spent Hydrocarbon in Keffi, Nasarawa State, Nigeria

Petroleum hydrocarbon is a major environmental pollutant throughout the world. This study was aimed at the effect of agro-wastes on bioremediation of soil polluted with spent hydrocarbon in Keffi. Hydrocarbon contaminated soils were collected from three (3) different auto mechanic workshops in Keffi, Nasarawa State Nigeria. The agro wastes used as stimulants were corncobs, fruits peels and melon shells. The hydrocarbon utilization was determined using gravimetric analysis. The hydrocarbon utilization bacteria were isolated and identified using standard microbiological method. The hydrogen ions (pH) of the contaminated soil sample were 4.5. pH of corncobs (CB) was 6.2, melon shells (MS) was 5.6 and fruits peels (FP) was 7.6. The value of total nitrogen for contaminated soil was 0.45 %, when amended with CB was 6.2%, MS was 1.26% and FP was 3.56%. Organic carbon in contaminated soil prior to study was 5.80 %, when amended with CB was 9.03%, MS was 7.55% and FP was 10.12%. The Phosphorous in contaminated soil was 8.41 mg/kg, when amended with CB was 13.40 mg/kg, MS was11.82mg/kg and FP was 15.61 mg/kg. Effect of time in reduction of hydrocarbon from contaminated soil amended with CB recorded highest after 40days (44.98 mg/kg) and the least (10.31 mg/kg) after 10days. Contaminated soil amended with CB the highest reduction was at 30 ℃ (48.92 mg/kg) and the least was at 35℃ (19.21mg/kg). From FP the highest reduction was at 30℃ (48.04 mg/kg). The highest degrading bacteria count was recorded from Keffi garage mechanic workshop (4.04 x106 cfu g/ml) and the lowest was from pyanko road mechanic workshop (1.18 x106 cfu g/ml). The highest bacteria isolated from none amended contaminated soil was Flavobacterium sp (33.3 %) and the lowest was Bacillus subtilis and Proteus sp (33.3%). From amended soil with CB the highest were Corynebacteria spp and Flavobacterium sp (100%) and the lowest were Bacillus subtilis, Pseudomonas flourescens, and Proteus sp (66.6%). From contaminated soil amended with MS the highest bacteria isolated were Flavobacterium sp and Corynebacteria sp (33.3%) and the lowest were Pseudomonas flourescens and Proteus sp (33.3 %). From contaminated soil amended with FP the highest bacteria isolated were Pseudomonas flourescens and Proteus sp (100%) and the lowest were Bacillus subtilis, Flavobacterium sp and Corynebacteria sp (66.6%). It is recommended that the environmental agencies in the country give consideration to agricultural waste products for bioremediation of hydrocarbon polluted soil in Nigeria

Application of Trichoderma sp in pineapple biomass composting

Background: Pineapple litter has great potential as an organic source and if it can be decomposed properly it can turn into organic fertilizer. Pineapple biomass is difficult to decompose due to high lignin and cellulose content which slow down the decomposition process. Thus the addition of auxiliary microbes such as decomposer fungi is thought to increase the rate of decomposition. Aim: The aim of this research is determine the effect of Trichoderma sp. inoculum grown on rice substrate on pineapple litter composting Method: This study was in a completely randomized design (CRD) method consisting of 6 treatments with two stages of testing, there are fungal inoculum productivity test and application on pineapple litter. The spore density and viability (CFU) values were calculated for the inoculum that was incubated for 14 days to determine the productivity level of the inoculum. The pineapple litter given the inoculum is then subjected to a decomposition process for seven weeks. Pineapple litter compost was analyzed at the fourth and seventh week with the parameters observed were C-organic content and C/N ratio. Result: The results showed the spore density was 2.0 x107 spores / ml and the CFU value was 10.6x108 cfu / ml with 100% viability. The best compost had the lowest C-organic and C/N ratio value, there are P3 compost obtained 12,5% C-organic and P2 compost obtained 9,6 C/N ratio. Conclusion: The use of rice media as an inoculum medium for the Fungi Trichoderma sp. can increase the level of productivity of Fungi Trichoderma sp. with a viability percentage of 100% at 14 days of inoculum age.

Boophilin D1, a Kunitz type protease inhibitor, as a source of inhibitors for the ZIKA virus NS2B-NS3 protease

Arboviruses are a global concern for a multitude of reasons, including their increased incidence and human mortality. Vectors associated with arboviruses include the mosquito Aedes sp., which is responsible for transmitting the Zika virus. Flaviviruses, like the Zika virus, present only one chymotrypsin-like serine protease (NS3) in their genome. Together with host enzymes, the NS2B co-factor NS3 protease complex are essential for the viral replication cycle by virus polyprotein processing. To search for Zika virus NS2B-NS3 protease (ZIKVPro) inhibitors, a phage display library was constructed using the Boophilin domain 1 (BoophD1), a thrombin inhibitor from the Kunitz family. A BoophilinD1 library mutated at positions P1–P4′ was constructed, presenting a titer of 2.9x106 (cfu), and screened utilizing purified ZIKVPro. The results demonstrated at the P1–P4’ positions the occurrence of 47% RALHA sequence (mut 12) and 11.8% RASWA sequence (mut14), SMRPT, or KALIP (wt) sequence. BoophD1-wt and mutants 12 and 14 were expressed and purified. The purified BoophD1 wt, mut 12 and 14, presented Ki values for ZIKVPro of 0.103, 0.116, and 0.101 μM, respectively. The BoophD1 mutant inhibitors inhibit the Dengue virus 2 protease (DENV2) with Ki values of 0.298, 0.271, and 0.379 μM, respectively. In conclusion, BoophD1 mut 12 and 14 selected for ZIKVPro demonstrated inhibitory activity like BoophD1-wt, suggesting that these are the strongest Zika inhibitors present in the BoophD1 mutated phage display library. Furthermore, BoophD1 mutants selected for ZIKVPro inhibit both Zika and Dengue 2 proteases making them potential pan-flavivirus inhibitors.

Assessment of Soil Properties Using Geospatial Techniques at DDUCE-OF Farm

The purpose of this study was to identify the geographic variability and mapping of different soil properties using geospatial techniques at DDUCE-OF Farm. For this purpose, surface soil samples (0-15 cm depth) were collected with the help of GPS at definite locations of research farm area at DDUCE-OF, CCSHAU, Hisar during rabi, 2021. These samples were analyzed in the laboratory for various physico-chemical and biological properties. Soil properties of DDUCE-OF farm revealed that soils are sandy loam to loam in texture having pH ranged from 7.15 to 8.65 and electrical conductivity (EC) ranged from 0.30 to 2.75 dS/m. The soil organic carbon (SOC) content was observed medium to high (0.41 to 0.94%).The available N was low (126 to 196 kg/ha), P ranged from 8.50 to 23.5 kg/ha (from low to high in content) and available K was found medium to high and ranged from 128.5 to 554.0 kg/ha. The available S ranged between 60 to 725 mg/kg and was observed sufficient. The soil microbiological bacterial count status ranged between 4.6x103-9.8x109 CFU count per g of soil in various blocks of organic farm. The DTPA-extractable micronutrients Zn (1.00-4.47 mg/kg), Fe (7.26-19.92 mg/kg), Mn (3.88-17.77 mg/kg) and Cu (0.93-4.64 mg/kg) were sufficient in amount and heavy metal contents (Ni, Pb, Cd, Cr, As) were found below their permissible limit. The study concluded that the soil mapping and survey is significant because it aids in the evaluation of soil qualities and their application in organic farming.

Influence of Azospirillum brasilense on micro-propagated sugarcane seedlings

Sugarcane is an essential crop in the Brazilian economy. The adoption of techniques in the acclimatization of plants in vitro contributes to success in the field. The present work aims to evaluate the effect of Azospirillum brasilense on the acclimatization of sugarcane seedlings. The experiment was conducted with six treatments and twelve replications. There were two control treatments without inoculation, one with N fertilization and one without, and four with A. brasilense (2.0x108 CFU) at different doses (19.92mL + N fertilization, 9.84mL, 19.92mL, and 39.84mL). Evaluations were seedling height (SH), stem diameter (SD), number of shoots per tube (NS), number of leaves (NL), fresh aerial weight (AFM), root fresh weight (RFM), total chlorophyll (TC), foliar nitrogen (FN) and visual microscopic study of the roots. The results showed that the recommended dose of A. brasilense + N fertilization increased by 36% in seedling height, 29% in stem diameter, 85% in aerial fresh mass, 21% in total chlorophyll, 142% in foliar N, and also increased root hairs and the presence of microbial biofilm. Therefore, the bacterium A. brasilense can be used in association with nitrogen fertilizers or alone, where it obtained great benefits in the increase of biomass.

EFFICACY OF SALINE WASH PLUS VANCOMYCIN/TOBRAMYCIN-DOPED PVA COMPOSITE (PVA-VAN/TOB-P) IN A MOUSE POUCH INFECTION MODEL IMPLANTED WITH 3D-PRINTED POROUS TITANIUM CYLINDERS

The efficacy of saline irrigation for the treatment of periprosthetic infection (PJI) is limited in the presence of infected implants. This study evaluated the efficacy of vancomycin/tobramycin-doped polyvinyl alcohol (PVA)/ceramic composites (PVA-VAN/TOB-P) after saline irrigation in a mouse pouch infection model.3D printed porous titanium (Ti) cylinders (400, 700 and 100 µm in pore size) were implanted into mice pouches, then inoculated with S. aureus at the amounts of 1X103 CFU and 1X106 CFU per pouch, respectively. Mice were randomized into 4 groups (n=6 for each group): (1) no bacteria; (2) bacteria without saline wash; 3) saline wash only, and (4) saline wash+PVA-VAN/TOB-P. After seven days, pouches were washed out alone or with additional injection of 0.2 ml of PVA-VAN/TOB-P. Mice were sacrificed 14 days after pouch wash. Bacteria cultures of collected Ti cylinders and washout fluid and histology of pouch tissues were performed.The low-grade infection (1X103 CFU) was more significant in 400 µm Ti cylinders than that in Ti cylinders with larger pore sizes (700 and 1000 µm (p<0.05). A similar pattern of high-grade infection (1X106 CFU) was observed (p<0.05). For the end wash, the bacteria burden (0.49±0.02) in saline wash group was completely eradicated by the addition of PVA-VAN/TOB-P (0.005±0.001, p<0.05).We noticed that 400 µm Ti cylinders have the highest risk of implant infection. Our data supported that the effect of saline irrigation was very limited in the presence of contaminated porous Ti cylinders. PVA-VAN/TOB-P was biodegradable, biocompatible, and was effective in eradicating bacteria retention after saline irrigation in a mouse model of low grade and high-grade infection. We believe that PVA-VAN/TOB-P represents an alternative to reduce the risk of PJI by providing a sustained local delivery of antibiotics.

The effectivity of probiotic containing Bacillus spp. applicated in the rearing of mud crab Scylla tranquebarica larvae to produce crablet

Probiotic bacteria in larvae rearing is expected to suppress pathogenic bacteria and improve water quality. The research aimed to determine the effectivity of probiotic bacteria containing Bacillus subtilis and B. licheniformis on larval rearing of purple mud crab Scylla tranquebarica to produce crablet. The larvae were stocked in nine units of conical fiber tank, fed Rotifers, Brachionus plicatilis, and nauplii Artemia sp. The probiotics tested, namely, A). RICA-4, containing the bacterium Bacillus subtilis, B). RICA-5 containing B. licheniformis, C). A combination of probiotics RICA-4 and RICA-5. The administration of each probiotic with a density of 108 CFU g−1 (5 mg L−1) was given to the water media for larvae rearing. The lowest nitrite and Total Organic Matter in the zoea stage were found in treatment B. At the megalopa stage, treatment B’s lowest TOM concentration occurred and was significantly different (p<0.05) from treatment C. On the other hand, the highest population of Vibrio spp (5.5x103 CFU mL−1) was obtained in treatment A. In contrast, treatment C had the highest bacteria population (4.8x104 CFU mL-1). On day 19, the megalopa in treatment B was more abundant than in treatments A and C causing crablet production in treatment B (280.5±47.50 ind. tank −1) was significantly higher (p<0.05) than crablet production in treatment C (196.5±29.50 ind. tank −1) and A (161.0±21.00 ind. tank −1). Therefore, improving water quality in treatment B may have an important factor in enhancing crablet production.

Real Time Paper Based Detection of Streptococcus bovis using Chromogenic Substrate in Resource Constrained Environments

Healthcare management faces a variety of difficulties in Circumstances with limited resources. Accurate diagnosis is the first step in disease prevention and therapy that works However, the diagnostic tools that are available in the economically advanced world are frequently of little utility in underdeveloped nations including Pakistan. The gram-positive bacterium Streptococcus bovis is an opportunistic pathogen that can cause various infections from superficial skin infections to severe and potentially fatal invasive diseases. Objective: To design a platform in environments with limited resources for the quick detection of Streptococcus bovis. Methods: A paper based analytical device (PAD) has been created with enrichment in sterile PYP broth for 1.5 hours and used for the detection of alkaline phosphatase activity using the chromogenic substrate Para-Nitrophenyl Phosphate (PNPP), the specie could be found in clinical samples. A coagulase test and sample Gram staining was conducted with the test. Qualitative detection was evaluated by visual detection while quantitative analysis is carried out using Image J software. Results: Alkaline Phosphatase ( S. bovis) reacts with the PNPP substrate (5.7 mM) in the presence of PAD. Concentrations below 4.5x104 cfu mL resulted in the detection of a color change. The micro PADs were incubated at 37 °C for 3–4 hours before reaction. Colored product (yellow) indicated the presence of S.bovis. Conclusion: Within 2 hours including enrichment time the test may identify Streptococcus bovis up to 104 CFUmL-1

Dose-response relationship of Ralstonia solanacearum and potato in greenhouse and in vitro experiments.

Ralstonia solanacearum is the causative agent of bacterial wilt of potato and other vegetable crops. Contaminated irrigation water contributes to the dissemination of this pathogen but the exact concentration or biological threshold to cause an infection is unknown. In two greenhouse experiments, potted potato plants (Solanum tuberosum) were exposed to a single irrigation with 50mL water (non-invasive soil-soak inoculation) containing no or 102 - 108 CFU/mL R.solanacearum. The disease response of two cultivars, Kondor and HB, were compared. Disease development was monitored over a three-month period after which stems, roots and tubers of asymptomatic plants were analyzed for latent infections. First wilting symptoms were observed 15 days post inoculation in a plant inoculated with 5x109 CFU and a mean disease index was used to monitor disease development over time. An inoculum of 5x105 CFU per pot (1.3x102 CFU/g soil) was the minimum dose required to cause wilting symptoms, while one latent infection was detected at the lowest dose of 5x102 CFU per pot (0.13 CFU/g). In a second set of experiments, stem-inoculated potato plants grown in vitro were used to investigate the dose-response relationship under optimal conditions for pathogen growth and disease development. Plants were inoculated with doses between 0.5 and 5x105 CFU/plant which resulted in visible symptoms at all doses. The results led to a dose-response model describing the relationship between R.solanacearum exposure and probability of infection or illness of potato plants. Cultivar Kondor was more susceptible to brown-rot infections than HB in greenhouse experiments while there was no significant difference between the dose-response models of both cultivars in in vitro experiments. The ED50 for infection of cv Kondor was 1.1x107 CFU. Results can be used in management strategies aimed to reduce or eliminate the risk of bacterial wilt infection when using treated water in irrigation.

The Influence of Storage on Structural Damage of Vegetable-Tanned Goat Skin by Microbes

This study aimed to determine the influence of storage on the structural damage of vegetable-tanned goat skin by microbes. The storage time was divided into three treatments, namely treatment P0 (0 months, storage), treatment P1 (6 months, storage), and treatment P2 (12 months, storage). The observation parameters for investigation included microbe contamination, pH levels, and levels of oil/fat. This study was conducted with Analysis Variance Completely Randomized Design (ANOVA) and Duncan Multiple Range Test (DMRT) to determine the differences between treatments. In contrast, analysis regression was used to obtain the optimum level. The research found that the highest amount of microbial contamination on the leather occurred at month 12, with the amount of microbial contamination reaching 4.46x103 CFU. The best storage for pH levels occurred at month 14 and pH 5.16. The skin with the best oil content is in the third month, with an oil/fat content of 7.97%.

Management of Postharvest Anthracnose Disease of Papaya (Carica papaya) Using Antagonistic Microorganisms

The antagonistic potential of microbial agents isolated from different sources like compost, rhizosphere soil, healthy fruits surface were investigated against Colletotrichum gloeosporiodes under In vitro and In vivo. The results of In vitro evaluation of 22 isolates of antagonists against Colletotrichum gloeosporiodes by dual-culture test showed Trichoderma harzianum UHSF-15 (with 62.50% inhibition of radial growth), Bacillus amyloliquifaciens UHSB-BCA-15 isolated from banana rhizosphere soil (50.00% inhibition) and Bacillus amyloliquifaciensUHSB-BCA-16 isolated from kiwi fruit surface (47.85% inhibition) to be most effective. In vivo screening of five effective microbial agentsselected from In vitro studies revealed that, Bacillus amyloliquifaciens UHSB-BCA-15to be most effective with lowest per cent disease index of 13.89% followed by Bacillus amyloliquifaciensUHSB-BCA-16 with 16.67% and carbendazium at 0.1% with 20.83% at 9d after storage. The higher per cent disease index of 58.33% was recorded in untreated control fruits. In another experiment where C. gloeosporoidesspore suspension (1x106 CFU mL-1) was challenge inoculated on fruits, the fruits treated with biocontrol agent Bacillus amyloliquifaciens UHSB-BCA-15 at concentrations of 1x108 CFU mL-1 for 5 minutes was found to be effective in reducing postharvest infection to a greater level with 33.33% disease index followed by Bacillus amyloliquifaciens UHSB-BCA-16 with 50.00% disease index and carbendazium at 0.1% with 52.77% disease index compare to control (91.66% disease index) at 9d after storage.

Ecosystem Biology of Soil Polluted with Spent Engine Oil

Soil communities, the microorganisms, are the initial and primary recipients of hydrocarbons or any xenobiotics incidented on the soil. This study evaluated the biology of soil organisms, ex-situ; the hydrocarbonclastics and non-hydrocarbonclastics, following an exposure to spent engine oil. The result demonstrated that at low and moderate contaminations of 1.5 and 2.5% w/w, hydrocarbons reduced the microbial population from control, 1.28 x109 cfu, to 3.42 x108 and 3.06 x 108cfu on week one respectively. However, at increased contaminations of 3.5% w/w, hydrocarbons from large oil spills weaken microbial ability to degrade the added hydrocarbons as the prejudicial nature reduced the population to 1.62 x 108cfu on week one. Overall, across the weeks, there was insurgence of hydrocarbon-degrading organisms, the hydrocarbonclastics, which on the other hand induced a limitation in microbial diversity. The isolated hydrocarbonclastic microorganisms in the descending order of their biodegradation potentials are: Pseudomonas aeruginosa, Arthrobacter, Alealigena, Corynebacter, Flavo-bacterium, Archromobacter, Micrococcus, Norcardia and Myco-bacterium. Thus within the ecosystem they devised metabolic diversity with increased catabolic properties primarily aimed at biodegrading the xenobiotics; making them hydrocarbonclastic organisms.