Advancements in Life Sciences (E-Journal, University of the Punjab)
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Characterization of antibiotic resistant gene in Staphylococcus aureus isolated from surgical wounds
Get PDFBackground: In the field of surgery wound infections have been a problem. Staphylococcus aureus, Escherichia coli, Proteus species, Klebsiella species, Streptococcus species, Enterobacter species, Pseudomonas species and species of staphylococci are frequently isolated from wound. This study was carried out to determine the prevalence of different pathogens in surgical wounds and their antimicrobial susceptibility patterns.Methods: Pus swabs from each patient was collected aseptically, and inoculated on culture media. Isolates were characterized, identified, and their antibiotic susceptibility patterns were determined using the Kirby-Bauer diffusion method.Results: Out of 300 surgical wound specimens analyzed, 208 samples were positive culture among which Staphylococcus aureus was most frequent pathogen with 103 (49.51%). The other 33 (15.86%) isolates were Pseudomonas aeruginosa followed by 27 (12.98%) isolates ofKlebsiella 25 (12.01%) isolates of Proteus and 20 (9.61%). isolates of E.coli respectively. The antibiotic susceptibility of Staphylococcus aureus was checked.Conclusions: It was observed that 103 isolates of S. aureus were resistant to Augmentin 65.04% to Sparfloxacin 59.22% to Amikacin 57.28% to Cefuroxime 57.28% to cefotaxime 56.31% to Ceftazidime 53.39% to Fusidic acid 48.54% to Ciprofloxacin 39.80% to Methicillin 37.86% to Gentamicin 28.15% to Meropenem 13.59% to Imipenem 5.82%. PCR assay for the detection of clinically relevant antibiotic resistance gene of S. aureus was done. Fragments of mecA (encoding methicillin resistance) gene were amplified and commercially sequenced which showed insertion at three sites; 480-481: T, 484-485: T, 464-465: G
Gene Profiling for Invertase Activity: Assessment of Potato Varieties for Resistance towards Cold Induced Sweetening
Get PDFBackground: Potato is the most important staple food in the world. Cold-induced sweetening occurs when potatoes are stored at low temperature for longer period of time. Due to non- enzymatic Millard reaction it causes unwanted changes in colour, taste and in flavor when fried and roasted at high temperature. However, long-term cold storage is mandatory to keep an adequate supply of potatoes throughout the year. The cause of cold-induced sweetening is invertase enzyme. Methods: Five potato varieties (Hermes (A) Lady Rosetta (B) Oscar (C) Kuroda (D) and Multa (E)) were investigated for invertase activity during two month cold storage at 4°C. Crude protein was extracted by PD Midi Trap G25 column technique. Quantification of mRNA expression was employed through QPCR. Determination of sucrose, reducing sugars and organic acids was simply done by 80% ethanol method and concentration were find out by using HPLC with already set standards. The correlation between invertase enzyme, sugar content and mRNA expression was calculated through Statistical methods.Results: Significant activity of invertase was observed at 4ºC with up to 6.3 nmol/min/mg of protein in the type-1 & 4 (cv. Hermes and Kuroda); 2.5 times less in type 2 (Rosetta) and 3.5 times less in type 3 (Multa) when compared with same at 4ºC. In addition, malic acid concentration was found positively correlated with invertase activity at 4ºC as compared with its concentration at harvesting time. However, citric acid and oxalic acid concentrations were independent of invertase enzyme activity. The transcript level of invertase enzyme was found significantly high in potato tubers stored at 4ºC in result 1 & 4 type, less in result type 2(C) and negligible in result 3(E) potato variety when revealed through reverse transcription PCR.Conclusions: In conclusion, Oscar (C) and Multa (E) were found more resistant to CIS at 4ºC storage and may be used for future variety improvement programs for CIS resistant through breeding and molecular approaches
Characterization of mercury resistant and growth promoting Enterobacter sp. from rhizosphere to use as a biofertilizer
Get PDFBackground: Mercury occurs naturally in environment, it is heavy metal that exists in three chemical forms like elemental mercury, inorganic mercury and organic mercury. All forms of mercury are problematic for living organism. Currently, the contamination of agricultural land and water systems with mercury has become one of the major environmental issues. The cheapest mode to remove mercury metal and its other forms from the ecosystem is the use of microorganisms.Methods: In this study, initially bacterial species were isolated and purified from nodule like structures on roots and stems of plants on MacConkey agar medium. Further screening for resistance to mercury was done on N- agar medium supplemented with different concentration of HgCl2 (20, 30, 40 and 50 µg/mL). Well plate method was used for the determination of bacterial strains having maximum ability to detoxify mercury. Selected bacterial strains were subjected to different biochemical tests for characterization and other metabolic tests were also performed to characterize their capabilities.Results: All strains were highly resistant to HgCl2 at the concentration of 20 µg/mL and moderately resistant at 30 µg/mL. Bacterial strain S-2 was moderately resistant and S-3 was least resistant at 40 µg/ml whereas S-2 was least resistant at 50 µg/ml. Selected bacterial strains were positive for nitrogen fixation and protease production, negative for phosphate solubilization but only S-1 was positive for hydrogen cyanide (HCN). Bacterial species were molecular characterized by 16S rDNA sequencing as Enterobacter cloacae (KJ857483, KJ857484 and KJ857485: NCBI GeneBank).Conclusion: Selected Enterobacter sp. exhibiting multiple characteristics can be used as biofertilizer in mercury polluted land for sustainable agriculture
Toxicity, analgesic and sedative potential of crude extract of soil-borne phytopathogenic fungi Aspergillus flavus
Get PDFBackground: Aspergillus flavus is one of the most abundant mold present around the world. The present study was conducted to investigate the acute toxicity, analgesic and sedative effect of the crude extract obtained from soil borne fungi A. flavus.Methods: The fungi was isolated from soil samples and identified morphologically and microscopically. The growth condition i.e. media, temperature, pH, and incubation period were optimized. In these optimized growth condition, A. flavus was grown in batch culture in shaking incubator. Crude contents were extracted by using ethyl acetate solvent. Crude secondary metabolites were screened for acute toxicity, analgesic and sedative effect.Results: Upon completion of the experiment, blood was collected from the tail vein of albino mice, and different haematological tests were conducted. White blood cells counts displayed a slight increase (10.6× 109/L) above their normal range (0.8–6.8 × 109/L), which may be due to the increment in the number of lymphocytes or granulocytes. However, the percentage of lymphocytes was much lower (17.7%), while the percentage of the granulocytes was higher (61.4%) than its normal range (8.6–38.9%). A reduction in the mean number of writhing in the different test groups was caused by the application of the crude ethyl acetate extract through the i.p. route at different doses (50, 100, and 150 mg/kg body weight). The results of our investigation showed the EtOAc extract of A. flavus can cause a significant sedative effect in open field.Conclusion: It was concluded from the present study that the A. flavus has the potential to produce bioactive metabolites which have analgesic and sedative effect
Successful DNA Profiling for Identification of burnt Families from their bones using AmpFℓSTR Identifiler® Plus Kit
Get PDFBackground: DNA profiling plays a vital role in the identification of dead bodies during mass disasters. Severe fragmentation, decomposition, burning and intermixing of the remains can occur in the mass disasters. DNA analysis faces many challenges especially when the dead bodies are completely decomposed or burnt. This report presents the identification of 32 completely burnt individuals including three families from their remains in a bus using AmpFlSTR Identifiler Plus® Kit and AmpFlSTR Y-filer® Kit.Methods: DNA was extracted from provided remains of burnt bodies and reference samples by organic extraction procedure. The extracted quantity of DNA was calculated on ABI SDS7500 real time PCR with Quantifiler® Human DNA Quantification Kit (Applied Biosystems). DNA samples of 32 completely burnt individuals including three families were amplified using AmpFlSTR Identifiler Plus® Kit and AmpFlSTR Y-filer® Kit. The genotyping of these amplified samples was performed on ABI 3130xl Genetic Analyzer.Results: The resulting data obtained from Genetic Analyzer was analyzed using GeneMapper ID software version 3.2 (Applied Biosystems). Seventeen burnt individuals including 3 burnt families were identified with the help of 16 autosomal STRs and 6 were identified through Y-STR analysis by allele sharing of their provided reference samples of parents and brothers respectively.Conclusion: For the identification of unknown individuals particularly burnt deceased victims, STR analysis has become the gold standard in forensic science. Successful DNA profiling through the amplification of STR markers of AmpFlSTR Identifiler Plus® Kit proved to be very helpful in identifying the remains of burnt individuals even in the presence of inhibition observed in the Real Time PCR
Is the war on terror induced-post traumatic stress disorder; the cause of suicide attack? An approach from psycho-cognitive and neurobiological perspective
Get PDFUnderstanding suicide attack is one of the highly complicated problems in the field of psychological disorders. Post-traumatic stress disorder (PTSD), may occur in individuals subjected to traumatic assaults like terrorism, warfare, sexual abuse or natural disaster. Individual's living within war affected area often develops PTSD, which may consequently leads to cognitive and memory impairment. The war induced PTSD patient, is under the influence of severe stress; terror and helplessness as it manipulate and retrieve the past trauma as a current threat. Substantial evidences support that PTSD patients are more prone to varying degree of neurological and psychological complications. In this correspondence, we wish to highlight the biological consequence of suicide attack from the perspective of war induced PTSD. Earlier research also supports that PTSD and suicide have some common basis like alterations in hypothalamic pituitary-adrenal axis, nitric oxide and catecholamine like norepinephrine and serotonin level. Thus it is important to uncover the risk of PTSD due to war on terror with precision towards suicide attack and minimize the detriment followed by it. PTSD through the development of depression, irritability and anger, is one amongst the major causes of suicide attack.In order to clarify the underlying psychological mechanism, there is a pressing need to address it from different aspects like disease causing synaptic plasticity and abnormal brain development. PTSD is a reaction to past traumatic events. For instance, the danger of perceived threat due to witness of deaths in a war, may develop a constellation of properties that may leads to PTSD. Usually, it initiates a sequence of behavioral and cognitive changes that can be anticipated to reduce the perceived threat. However, the consequences of perceived threat lead to cognitive changes and thus maintain a devastating disorder. Appraisals of such memory not only generate situational fear but also the avoidance, which leads to enhanced trepidation and over-activity. For example, a person exposed to a road side traumatic accident may avoid driving; for having an impractical faith that it may happen again, thereby affecting its social life.Patients suffering from PTSD due to war may interpret that the trauma will persist for long time and thus he is no longer safe. He or she may suffer from depression, irritability, anger outbursts, emotional numbing, flashbacks, and nightmares [1]. It has been reported that numbing is an ordinary reaction to traumatic events. Individuals should realize that it is a normal aspect of the recovery process, otherwise it can lead to permanent changes which may worsen their physical or psychological well-being [2-4]. Children experiencing the PTSD, usually underwent alterations in hypothalamic-pituitary-adrenal axis, catecholamine and norepinephrine, which results in pathological and detrimental brain development [5]. Interestingly, the inhibition of nitric oxide in hippocampus by antidepressant has promising outcome to alleviate the PTSD symptoms [6]. While, the augmented level of plasma nitrates in depressive patients are found to be associated with suicide attempts [7].Figure 1The intention of a suicide attack is to kill a large crowd or bringing mass destruction, even with the notion that he will die in this act. Suicide bombings termed as "suicide bombing" constitute an overall 4% of terrorist attacks, which dates back to the beginning of the 19th century. In most of the modern suicide terrorist is used against non-combatants for the accomplishment of impact on political situation. Although a suicide attack aims to annihilate a primary target, however it can be used as a weapon of psychological warfare to affect the large public population. The main target of this action is not those who are killed but those witnessed it [8]. Intriguingly, it is reported that the level of brain-derived neurotropic factor (BDNF) is directly associated with suicide while indirectly it can affect PTSD. Also, defining body of research proved that altered level of serotonin in dorsal raphe nucleus, amygdala, median raphe, frontal cortex, hypothalamus and hippocampus is associated with aggressive behavior and suicide [9]. PTSD-induce symptoms like impulsivity, violence, suicide attempts, depression, panic, and anxiety can also be associated with altered serotonin levels [10].ConclusionThe mechanism of suicide attack is still highly debated; and need much more to address. Since, PTSD through the development of depression, irritability and anger, accomplish various physiological and cognitive changes in the brain, so it might be one of the causes which increases the susceptibility of acceptance for being a volunteer to suicide attack? This discussion was put forward; as some war induced PTSD patients among temporary displaced people in different regions of the world showed strong willingness for suicide attack as a counter revenge of war. Therefore, we also need to address suicide attack from the perspective of psychological disorders like war induced PTSD
Effects of chronic mild stress on apomorphine induced behavioral sensitization in different brain regions of rats in relation to serotonin change
Get PDFBackground: The impacts of unpredictable stressors have influence on neurochemical and behavioral parameters in laboratory animals. Stress induced behavioral changes particularly those associated with anxiety like behavior may activate topographically organized mesolimbic cortical serotonergic system. This study was designed to investigate the influence of unpredictable stress on behavioral and neurochemical parameters in apomorphine treated rats.Methods: Initially, the animals were divided into two groups as Unstressed and stressed (uncontrollable chronic mild stress or UCMS). Both groups of animals were subdivided into two groups; i.e. saline and apomorphine administrated animals at dose 1.0 mg/kg. Behavioral manipulations was observed by monitoring the locomotor activity and exploratory activity. Neurochemical estimation of 5-hydroxytryptamine (5-HT) was done by High performance liquid chromatography (HPLC). Animals were decapitated 24hr post apomorphine injection and different regions of brain (dorsal and ventral striatum), of animals were collected and stored at -70°C.Results: This preclinical study showed that the UCMS induced hypophagia were promoted in apomorphine administrated animals. Apomorphine induced hyperlocomotion were more prominent in unstressed animals than that of stressed groups. It implies that apomorphine is effective in the retrieval from UCMS induced depressive symptoms in rats. Neurochemical study showed decreased level of 5-HT in unstressed animals than stressed animals in response to apomorphine administration.Conclusion: This study, therefore establish the relation between stress and addiction at behavioral as well as neurochemical level to better understand the idea whether intolerable stress promotes addiction
Detection of bacterial load in drinking water samples by 16s rRNA ribotyping and RAPD analysis
Get PDFBackground: Safe and healthy drinking water is inaccessible to more than 20% of the world population. Among some major risks to safety of potable water, contamination with pathogenic microorganisms is the most alarming and harmful Therefore, it is needed to develop and implement fast and accurate methods for the detection of bacterial contamination in water. Methods: Biological analysis of drinking water samples obtained from nine different collection points of Lahore city was carried out and total of six different bacterial strains were isolated. Biochemical characterization was done under standard laboratory conditions. Molecular identification of these isolates was done by using random amplified polymorphic DNA (RAPD) analysis. Results: The drinking water sample collected from Punjab University showed highest bacterial count 1066/0.5 ml of drinking water while residential area of University of the Punjab contained least number of bacterial counts i.e., 38/0.5 ml of drinking water. Amplification patterns of isolates SZ1, SZ3, SZ4 and SZ6 obtained by RAPD were found similar to genus Bacillus. While, SZ2 and SZ5 had unique amplification patterns identical to Bacillus megaterium. All the six bacterial strains were tested for the presence of protease, lipase, cellulase, and amylase. Strain SZ2 gave positive result for all of them except amylase.Conclusion: Tube well water of Punjab University area of Lahore is safe for drinking purpose except admin block tube. It is recommended to monitor the bacteriological load of drinking water at regular intervals in order to control water borne bacterial diseases
Phytochemical Analyses for Antibacterial Activity and Therapeutic Compounds of Convolvulus arvensis L., Collected from the Salt Range of Pakistan
Get PDFBackground: Pharmaceutical world is currently facing a major issue of bacterial resistance against antibiotics. For the past few years, plants being an excellent source of phytochemicals have replaced many traditional antibacterial agents. Keeping this in view, the current study was conducted to detect the antibacterial activity and the presence of various phytochemicals in Convolvulus arvensis leaves and stem extracts.Methods: Eight different organic and aqueous extracts of Convolvulus arvensis L. (C. arvensis) leaves and stem were prepared and their antimicrobial activity against 13 clinically important bacterial species was investigated using the disc diffusion assay. Phytochemical screening of the aqueous extracts of C. arvensis was performed using qualitative tests. Results: All the thirteen bacterial species tested in this study were found sensitive to the stem and leaf extracts of C. arvensis. Highest antimicrobial activity was observed against Escherichia coli while among all the extracts butanol extract was the most effective antimicrobial agent. Phytochemical analyses using organic and aqueous extracts confirmed the presence of various secondary plant metabolites of therapeutic value i.e. saponins, flavonoids, steroids, tannins, phlobatannins, cardiac glycosides and coumarins.Conclusion: Our results indicate that C. arvensis plant contains some phytochemicals with antimicrobial affects. These chemicals possess great potential to be used as advanced therapeutic compounds against broad range of pathogenic bacteria
Estimation of Different Biochemical Intensities in Drinking Water from Eastern Region of Lahore City
Get PDFBackground: Lahore is city of over 8 million population with consumption of over 350 million gallons of fresh water per day. The present study was undertaken to determine the suitability of ground water from different areas of Mughalpura Lahore which is being used for drinking purposes.Methods: The ionic concentration of TDS (total dissolved solids), TSS (total suspended solids), calcium, magnesium, sulphate, chloride, carbonate, bicarbonate and alkalinity in the collected water samples was determined. The concentration of heavy metals like Cu, Cr, Zn, Cd, As, Pb and Fe was estimated by applying American Society for Testing and Materials (ASTM) methods. The EC (electrical conductivity), pH, BOD (biological oxygen demand) and COD (chemical oxygen demand) values of the water were also measured. Data was statistically analyzed through analysis of variance technique.Results: Increasing concentrations of heavy metals like Cu, Cr, Zn, Cd, As, Pb, Fe and TDS, TSS, calcium, magnesium, sulphate, chloride, carbonate, bicarbonate and alkalinity were obtained. The observed values of EC, BOD and COD were also high in range. Only the pH value obtained was in the range of WHO standards.Conclusion: It was concluded from correlation among various chemical components in water like Ca+2, Mg+2, HCO3- and SO4-2 that this water is not safe for drinking according to WHO standards. The correlation among alkalinity, pH, Ca+2, Cl-, EC, HCO3-, SO4-2, TH and TSS further suggested that the samples with higher amount of these components must not be used for drinking purposes by animals and human