Acute Post-Traumatic Locked Knee - An Unmasking of a Rare Knee Disorder

@#Locked knees are commonly caused by meniscal tears, floating osteochondral bodies, ruptured anterior cruciate ligament (ACL) stump, or other mechanical origins in the knee. Some locked knees occur spontaneously, while in most cases, by a preceding knee trauma. Locked knees are rarely caused by a pathological growth in the knee. More unusually is the occurrence of locked knee caused by a pre-existing pathological entity after a traumatic event. We report a rare case of locking in the knee by a pre-existing knee condition presented only after trauma to the knee. This case emphasizes that locking in the knee can be caused by a pathology that may be asymptomatic until it is revealed by a traumatic event.

Detection of Pathogenic Leptospira in Rats and Phylogenetic Analysis Using Outer Membrane Lipoprotein Lipl32 Gene at Two Major Public Markets

@#Introduction: Limited studies have been documented on the presence of pathogenic Leptospira in public markets serving the community in sub-districts of Selangor. The aim of this study was to detect the presence of pathogenic Leptospira in rats using a gene encoding an outer membrane lipoprotein LipL32. Methods: Polymerase chain reaction (PCR) was performed using LipL32 primers on sixty kidney samples of rats trapped at two locations of study; Pasar Borong Selangor in Seri Kembangan and Pasar Basah Bandar Baru Bangi in Bangi. Results: Out of 60 samples analysed, 36.7% were positive for the presence of LipL32. All positive samples highly matched (>94%) nucleotide sequence for LipL32 of pathogenic Leptospira and related to the pathogens through phylogenetic analysis. Conclusion: The detection of LipL32 indicates the potential presence of pathogenic Leptospira species at public markets. Although only 60 rats were successfully trapped, the rats are mobile and might further transmit the pathogenic organisms to other areas.

Assessment of Pathogenicity of Community-Acquired MRSA Isolates in Mice-Induced Peritonitis

@#Introduction: Methicillin-Resistance Staphylococcus aureus (MRSA) is known as a major nosocomial pathogen in healthcare. However, it has now spread in the community known as community-acquired MRSA (CA-MRSA). Thus, the survival and pathogenicity of CA-MRSA isolates were assessed using in vivo peritonitis model with comparison to ATCC-MRSA. Two CA-MRSA isolates; CA-MRSA1 and CA-MRSA2 that were isolated from healthy population, were studied and compared. Methods: Mice were assigned into 4 groups and injected intraperitoneally with ATCC-MRSA, CA-MRSA1 or CA-MRSA2, respectively. Sterile Dulbecco’s Phosphate-Buffered Saline (DPBS) represents negative control. Mice were observed twice daily, 0-72 hours of post-infection. Any signs of distress were recorded for severity score and survival analyses. Mice were euthanised at 72 hours post-inoculation or by referring to the Peritonitis Severity Scoring (PSS) system. Organs of interest, peritoneal lavage and abscess were processed for bacterial counts. Tissue samples were analysed for histopathological scores. Results: All mice inoculated with MRSA showed clear signs of illness with peritonitis symptoms of p<0.001 and comparable PSS scores were recorded in all infected mice groups. Intraperitoneal injection of lethal dose of MRSA resulted in significant death of ATCC-MRSA (p<0.05) and CA-MRSA-infected mice (p<0.01), compared to the un-infected. Bacterial burden was significantly high in all samples harvested from mice challenged with CA-MRSA2 compared to ATCC-MRSA except in abscess and lung. Significant liver necrosis and spleen inflammation were observed in CA-MRSA1, and lung inflammation in ATCC-MRSA-infected mice. Conclusion: Nasal carriage CA-MRSA isolates from a healthy population has the potential to cause peritonitis with comparable severity as ATCC-MRSA.

Proteomic profiling of Acinetobacter baumannii ATCC 19606 and Malaysian isolated clinical strain AB-13 using 2-dimensional gel electrophoresis

Aims@#Acinetobacter baumannii has been identified as one of the six most pathogenic bacteria that is the cause of most hospital bacterial infections according to Infectious Disease Society of America (IDSA). These nosocomial pathogens are notorious worldwide due to its ability in causing lethal infections among immunocompromised patients and its resistance to many strong antibiotics. This study aims to compare the expressed proteins of two A. baumannii strain, ATCC 19606 and a pathogenic clinically isolated strain known as AB-13. @*Methodology and results@#AB-13 clinically strain was isolated from the lower respiratory tract of a patient with pneumonia. In this study, the proteomic profile of both ATCC 19606 and AB-13 are produced using 2-dimensional gel electrophoresis. The total protein contents were extracted, quantified and separated using 2-DE with a pH range of 4-7 to acquire the proteomic profile for comparison. The final analytical gel was analysed using Delta2D software and among the 324 protein spots successfully resolved, 10 spots exhibited signs of differential expression with 7 spots found to be downregulated and 3 spots upregulated (p< 0.01). These differences could signify the evolution AB-13 has undergone as it acquires traits ultimately aiding in its survivability, antimicrobial resistance and pathogenicity within varied environments especially during infections.@*Conclusion, significance and impact of study@#These findings support the presence of variation in AB-13 from a proteomic perspective, highlighting the pathogen’s evolution improving survivability and pathogenicity, warranting in-depth exploration towards understanding A. baumannii virulence and pathogenicity.

Distribution of virulence genes and the molecular epidemiology of Streptococcus pyogenes clinical isolates by emm and multilocus sequence typing methods

@#Background: Streptococcus pyogenes has a variety of virulence factors and the predominant invasive strains differ according to specific emm types and geographical orientation. Although emm typing is commonly used as the gold standard method for the molecular characterisation, multilocus sequence typing (MLST) has become an important tool for comparing the genetic profiles globally. This study aimed to screen selected virulence genes from invasive and non-invasive clinical samples and to characterise the molecular epidemiology by emm typing and MLST methods. Materials and Methods: A total of 42 S. pyogenes isolates from invasive and non-invasive samples collected from two different tertiary hospitals were investigated for the distribution of virulence factors and their molecular epidemiology by emm and multilocus sequence typing methods. Detection of five virulence genes (speA, speB, speJ, ssa and sdaB) was performed using multiplex polymerase chain reaction (PCR) using the standard primers and established protocol. Phylogenetic tree branches were constructed from sequence analysis utilised by neighbour joining method generated from seven housekeeping genes using MEGA X software. Results: Multiplex PCR analysis revealed that sdaB/speF (78.6%) and speB (61.9%) were the predominant virulence genes. Regardless of the type of invasiveness, diverse distribution of emm types/subtypes was noted which comprised of 27 different emm types/subtypes. The predominant emm types/subtypes were emm63 and emm18 with each gene accounted for 11.8% whereas 12% for each gene was noted for emm28, emm97.4 and emm91. The MLST revealed that the main sequence type (ST) in invasive samples was ST402 (17.7%) while ST473 and ST318 (12% for each ST) were the major types in non-invasive samples. Out of 18 virulotypes, Virulotype A (five genes, 55.6%) and Virulotype B (two genes, 27.8%) were the major virulotypes found in this study. Phylogenetic analysis indicated the presence of seven different clusters of S. pyogenes. Interestingly, Cluster VI showed that selected emm/ST types such as emm71/ST318 (n=2), emm70.1/ST318 (n=1), emm44/ST31 (n=1) and emm18/ST442 (n=1) have clustered within a common group (Virulotype A) for both hospitals studied. Conclusion: The present study showed that group A streptococcci (GAS) are genetically diverse and possess virulence genes regardless of their invasiveness. Majority of the GAS exhibited no restricted pattern of virulotypes except for a few distinct clusters. Therefore, it can be concluded that virulotyping is partially useful for characterising a heterogeneous population of GAS in hospitals.

Sociodemographic and Psychological Factors as Predictors of Body Mass Index-for-Age (BAZ) among Adolescents in Sibu, Malaysia

@#Introduction: Understanding childhood obesity becomes vital as a tremendous increase in the prevalence of overweight/obesity among children and adolescents was observed. This cross-sectional study aimed to investigate the associations between sociodemographic and psychological characteristics with body-mass-index-for-age (BAZ) among adolescents in Sibu, Sarawak. Methods: A total of 375 students (32.0% males and 68.0% females; 15-17 years old) at four randomly selected public secondary schools were recruited. A questionnaire on sociodemographic and psychological characteristics (self-efficacy for physical activity, weight management, and nutrition, body discrepancy score, and sociocultural pressure to be thin) were used to gather information. Body weight and height were also assessed. Results: Around 18.6% respondents were found to be overweight/obese while nearly 5.0% were categorized as thin. In multiple linear regression, three significant predictors, namely body discrepancy score, being Iban (Reference: Chinese) and sociocultural pressure to be thin explained 45.1% of the variance in BMI-for-age z-score. Conclusion: Future interventions on adolescent body weight management should consider incorporating sociodemographic and psychological factors such as the development of positive body image, uniqueness in cultural value, and management of perceived sociocultural pressures to increase their effectiveness

Multilocus Sequence Typing Analysis of Invasive and Non-Invasive Group B Streptococcus of Hospital Origin in Malaysia

@#The aim of this study was to study the genotype of a hospital collection of Group B Streptococcus (GBS) from invasive and non-invasive sites. Fifty-one pre-characterised human of GBS were re-identified and further analysed by multilocus sequence typing (MLST) in relation to previously published serotypes. Fifteen sequence types (ST) were found with ST1 being the most predominant. ST1 was also associated with majority of the invasive isolates. The genotypic distribution patterns of GBS in this study were largely in agreement with previous reports from other countries indicating the tendency of certain genotypes to prevail in human infection settings.

The PUTRA Press-fit System as An Alternative Femoral Fixation Technique for Posterior Cruciate Ligament Reconstruction – A Technical Note and Outcome Evaluation

Abstract@#We describe our technique of single-bundle PCL reconstruction using a bony femoral press-fit system. 9 patients underwent PCL reconstruction using our pressfit system. The surgical procedure is described in detail. Post-operatively, 5 patients were available for assessment and review. Four patients gave a final Lysholm score of 92 – 100%. Three patients gave a Hospital for Special Surgery (HSS) score of excellent, one fair and one poor. Two patients gave a reduced Tegner activity score post-operatively while the other three gave a similar score post-operatively. Assessment using KT-1000 revealed four patients with a side-to-side difference of less than 3 mm (average side-to-side difference, 1.87 mm), while one patient exhibited a side-to-side difference of 5.8 mm. We believe that our technique enhances tunnel healing through usage of a bone-plug fixation and provides a cheap alternative for graft fixation on the femoral side in PCL reconstruction.

Genotypic and phenotypic characterization of methicillin resistance determinants and β-lactamase in Staphylococcus species

Aims: To characterize the genotypic distribution of mec complex, bla complex, methicillin-resistance level (cefoxitinMIC) and β-lactamase activity in carriage methicillin-resistant Staphylococcus species for a potential correlation. Methodology and results: Biochemical test, 30 µg cefoxitin diffusion disc test, cefoxitin E-test, mec and bla complexes distributions, Pbp2a and β-lactamase assays were conducted to characterize phenotypic and genotypic of MRSA and MRCoNS in our collection. Phylogenetic tree was constructed using MEGA6 software to trace the diversity of blaZ gene of MRSA and MRCoNS. Sixteen MRSA and nineteen MRCoNS were identified by biochemical tests followed by 30 µg cefoxitin antibiotic disc susceptibility test and mecA gene screening. Twenty nine isolates carry complete mecA genes (2.1 kb), incomplete mec regulator (negative or truncated) and positive Pbp2a assay for both MRSA and MRCoNS. Only MRCoNS SC177 isolate with cefoxitin MIC of 32 µg/mL carries complete mec complex. Thirty-one of thirty-five isolates carry complete bla complex (blaZ, blaRI, blaI) with 10 MRSA produce strong β-lactamase and cefoxitin MIC of ≥12 µg/mL. Only 4 MRCoNS with cefoxitin MIC of ≤8 µg/mL produce strong β-lactamase. The diversity of blaZ gene was demonstrated by phylogenetic analysis and unusual amino acid mutation at position 145 for MRSA SA60 isolate may compromise its β-lactamase activity with low cefoxitin MIC level (2 µg/mL). Conclusions, significance and impact of the study: Isolates that carry complete complete mecA gene were largely consistent with the expression of Pbp2a. Nevertheless, there is no clear correlation of mec regulator genes in relation to cefoxitin-MIC in both methicillin resistant (MR) Isolates that carry Staphylococcus species. On the other hand, various expression level of β-lactamase may correlate with cefoxitin-MIC level in MRSA as compared to MRCoNS.

Possible mechanisms of antinociception of methanol extract of Melastoma malabathricum leaves

ABSTRACT Melastoma malabathricum L., Melastomaceae, has been traditionally used to relieve diverse pain-related ailments. The objectives of the present study were to determine the antinociceptive activity of methanol extract of M. malabathricum leaves and to elucidate the possible mechanisms of antinociception involved using various rats' models. The extract (100, 250, and 500 mg/kg) was administered orally 60 min prior to subjection to the respective test. The in vivo acetic acid-induced abdominal constriction, formalin-induced paw licking, and hot plate tests were used as the models of nociception to evaluate the extract antinociceptive activity. Further studies were carried out to determine the role of opioid and vanilloid receptors, glutamate system and nitric oxide/cyclic guanosine phosphate (NO/cGMP) pathway in modulating the extract antinociceptive activity. From the results obtained, M. malabathricum exhibited significant (p < 0.05) antinociceptive activity in all the chemical- and thermal-induced nociception models. Naloxone (5 mg/kg), a non-selective opioid antagonist, failed to significantly affect the antinociceptive activity of MEMM when assessed using the abdominal constriction-, hot plate- and formalin-induced paw licking-test. M. malabathricum also significantly (p < 0.05) reversed the nociceptive response in capsaicin- and glutamate-induced paw licking test. Furthermore, only L-arginine (a nitric oxide precursor) alone, but not, NG-nitro-L-arginine methyl esters (L-NAME; an inhibitor of NO synthase), methylene blue (MB; an inhibitor of cGMP), or their combination thereof, significantly (p < 0.05) block the antinociceptive activity of M. malabathricum. In conclusion, M. malabathricum exerted a non-opioid antinociceptive activity at the central and peripheral levels partly via the inhibition of vanilloid receptors and glutamatergic system, and activation of the NO-mediated/cGMP-independent pathway.

The emergence of Acinetobacter baumannii ST-6 multidrug resistant: A resilient biofilm producer, Malaysia

Aims: The aim of this research is to explore the presence of multidrug-resistance (MDR) Acinetobacter baumannii strains isolated from hospitalized patients in a tertiary-care center, Subang Jaya, Selangor, Malaysia and to compare their genotypic and phenotypic characteristics. Methodology and results: Clonal relationships were determined by multilocus sequence typing (MLST) and biofilm forming ability was evaluated by using 2, 3 - bis (2 - methoxy - 4 - nitro - 5-sulfophenyl) - 5 - [(phenylamino) carbonyl] - 2H-tetrazolium hydroxide (XTT) reduction assay in microplates and Congo red agar method (CRA). Four virulence genes coding for A. baumannii pilus usher-chaperone assembly protein, csuE gene; outer membrane protein, ompA gene; biofilm poly-β-1, 6-Nacetylglucosamine (PNAG) synthesis protein, pgaA gene; and acinetobactin-mediated iron acquisition protein, bauA gene were searched for in a collection of strains. Antimicrobial resistance against 11 antibiotics were studied by broth microdilution method. Seventeen A. baumannii clinical strains were isolated and MLST showed that the strains belonged to 5 distinct sequence types (STs), namely, ST-6, ST-265, ST-324, ST-325 and ST-432. Fiftythree percent of the strains were resistant to 4 or more antibiotics. Twelve strains produced biofilm and out of them, 4 were strong biofilm producer, besides, these strong biofilm producers were MDR strains and belongs to ST-6. In addition, all strains were ompA positive, biofilm producing strains were csuE and pgaA positive and only strong biofilm producing strains were bauA positive. Conclusion, significance and impact study: Our study demonstrates that the ST-6 strains in Malaysia could represent MDR, capable of forming strong biofilm and possess csuE, ompA, pgaA and bauA genes, virulence characteristics that probably help the bacteria to persist and cause infection.

Arthroscopic Mumford Procedure Utilizing the Anteromedial and Neviaser Portals – A Pilot Cadaveric Study on Neurovascular Structures at Risk

Introduction: Degenerative disorder involving the acromioclavicular joint (ACJ) is quite common especially in the elderly. One of the surgical modalities of treatment of this disorder is the Mumford Procedure. Arthroscopic approach is preferred due to its reduced morbidity and faster post-operative recovery. One method utilizes the anteromedial and Neviaser portals, which allow direct and better visualization of the ACJ from the subacromial space. However, the dangers that may arise from incision and insertion of instruments through these portals are not fully understood. This cadaveric study was carried out to investigate the dangers that can arise from utilization of these portals and which structures are at risk during this procedure. Materials and Methods: Arthroscopic Mumford procedures were performed on 5 cadaver shoulders by a single surgeon utilizing the anteromedial and Neviaser portals. After marking each portals with methylene blue, dissection of nearby structures were carried out immediately after each procedure was completed. Important structures (subclavian artery as well as brachial plexus and its branches) were identified and the nearest measurements were made from each portal edges to these structures. Results: The anteromedial portal was noted to be closest to the suprascapular nerve (SSN) at 2.91 cm, while the Neviaser portal was noted to be closest also to the SSN at 1.60 cm. The suprascapular nerve was the structure most at risk during the Mumford procedure. The anteromedial portal was noted to be the most risky portal to utilize compared to the Neviaser portal. Conclusion: Extra precaution needs to be given to the anteromedial portal while performing an arthroscopic distal clavicle resection in view of the risk of injuring the suprascapular nerve of the affected limb.

E-stamping: growth, opportunities and challenges in India.

Indian financial industry is witnessing a revolution. It is not just the metro cities and urban areas that are fueling the online scene in India but the tier 2-3 cities where there is internet penetration but non availability of electronic financial facility is also adding to growth.The growth of internet and other related technologies has enabled a new format called E-stamping to emerge. The e- stamping definition includes using the internet way to pay non judicial Stamp to the government. Estamping is short for electronic stamping and has been active on the internet as early as 2008. G2C (government to consumer) is another way e- stamping is described. The Government of India has mandated the Stock Holding Corporation of India (SHCIL) as central record keeping agency for e-Stamping. The CRA will also appoint authorised collection centres (ACC) & travelling vendor who issue certificate to their clients at their counters. The authorised collection centre would be Bank & Financial institutions, law firms, chartered accountant firms, professionals. E stamping has established itself as a viable option for old traditional stamping. This paper attempts to throw some lights on the growth, opportunities and challenges for e stamping in Indian context.

Exceptional Case of Omphalocele (Exomphalos Major).

An unregistered case of a 27-year-old multipara with 8 years married life was admitted on 3/11/09 at 10.30 pm with complaints of tightness of abdomen, backache and draining per vagina and delivered a fetus of 19 weeks in cephalic presentation with omphalocele (i.e., stomach, liver, bowel and bladder protruding into the proximal part of umbilical cord) on 5/11/09 at 9.00 pm at Sameena Maternity and Nursing Home, Hyderabad, Andhra Pradesh.

Effect of aqueous extract of Dicranopteris linearis leaves against paracetamol and carbon tetrachloride-induced liver toxicity in rats

The present study aimed to determine the hepatoprotective activity of Dicranopteris linearis L. [family Gleicheniaceae] leaf aqueous extract [DLAE] using two models of liver injury in rats. Rats were divided into ten groups [n=6] and received dH2O [negative control], 200 mg/kg silymarin [positive control] or DLAE [50, 250 and 500 mg/kg] orally once daily for 7 consecutive days and on the 8th day subjected to the hepatotoxic induction either using carbon tetrachloride [CCl[4]] or paracetamol [PCM]. The bloods and livers were collected and subjected to biochemical and microscopical analysis. From the data obtained, only the highest dose of DLAE significantly [p<0.05] reduced the ALP, ALT and AST levels in CCl[4]-and PCM-induced hepatotoxic rats while the other doses caused significant [p<0.05] reduction only in the levels of ALT and AST. The histological results obtained were in line with the biochemical analysis wherein reduction in the CCl[4]- and PCM-induced tissue formation of necrosis, steatosis and inflammation occurred in a dose-dependent manner. In conclusion, the DLAE possesses hepatoprotective activity, which could be attributed to its free radicals scavenging and antioxidant activities, and high flavonoids content. Thus, in-depth studies regarding the hepatoprotective activity of DLAE are warranted

Stages of change to increase fruit and vegetable intake and its relationships with fruit and vegetable intake and related psychosocial factors

BACKGROUND/OBJECTIVES: Understanding individual's intention, action and maintenance to increase fruit and vegetable intake is an initial step in designing nutrition or health promotion programs. This study aimed to determine stages of change to increase fruit and vegetable intake and its relationships with fruit and vegetable intake, self-efficacy, perceived benefits and perceived barriers. SUBJECTS/METHODS: This cross-sectional study was conducted among 348 public university staff in Universiti Putra Malaysia. A pre-tested self-administered questionnaire and two days 24-hour diet recall were used. RESULTS: Half of the respondents (50%) were in preparation stage, followed by 43% in action/maintenance, 7% in pre-contemplation/contemplation stages. Respondents in action/maintenance stages had significantly higher self-efficacy (F = 9.17, P < 0.001) and perceived benefits (F = 5.07, P < 0.01) while respondents in pre-contemplation/contemplation and preparation stages had significantly higher perceived barriers (F = 4.83, P < 0.05). Perceived benefits tend to outweigh perceived barriers pre-ceding to taking action. Self-efficacy is important in motivating individuals to increase fruit and vegetable intake as self-efficacy and perceived barriers crossed over between preparation and action/maintenance. Respondents in action/maintenance stages had the highest adjusted mean serving of fruit and vegetable intake (F = 4.52, P < 0.05) but the intake did not meet recommendation. CONCLUSION: Intervention strategies should emphasize on increasing perceived benefits and building self-efficacy by providing knowledge and skills to consume a diet high in fruits and vegetables in order to promote healthy changes in having high fruit and vegetable intake.

Prevalence of macrolide resistance and in vitro activities of six antimicrobial agents against clinical isolates of Streptococcus pneumoniae from a multi-center surveillance in Malaysia

The in vitro activities of 6 antimicrobial agents against clinical isolates of Streptococcus pneumoniae (pneumococci) were investigated and the erythromycin minimum inhibitory concentrations (MICs) were correlated with the two major macrolide resistance determinants, mef(A) and erm(B). MICs of commonly used antibiotics as well as the presence of macrolide resistance determinant genes in all isolates were tested. Seventy one pneumococcal isolates collected at Institute for Medical Research (IMR) were included in this study. Phenotypic characterization, MIC determination using E-test strips and polymerase chain reactions for antibiotic resistance determination were included. Among the isolates, 25 (35.2%) isolates were erythromycin susceptible, 3 (4.2%) were intermediate and 42 (60.6%) were resistant. Fifty three isolates (74.7%) were found with mef(A) alone, 15 (21.1%) isolates with erm(B) + mef(A) combination and 3 (4.2%) isolates with none of the two genes. The in vitro activity of penicillin, amoxicillin clavulanic acid, ceftriaxone and cefotaxime is superior to trimethoprim-sulfamethoxazole and erythromycin. In conclusion, pneumococcal isolates in this study were highly susceptible to penicillin with very low MICs. However, a very high prevalence rate of erythromycin resistance was observed. Erythromycin resistant S.pneumoniae isolates with both mef(A) and erm(B) showed very high MICs ≥256 μg/mL.

Staphylococcus aureus Nasal Carriers Among Medical Students in A Medical School

Staphylococcus aureus is usually considered a colonizer but can result in infections under favourable conditions, especially in the healthcare setting. Healthcare workers can be colonized by S. aureus, and may transmit them to patients under their care. We conducted a cross sectional study to determine the prevalence of S. aureus nasal carriers among medical students in Universiti Putra Malaysia (UPM) (from January to June 2011). Our study involved 209 medical students comprising of 111 and 97 preclinical and clinical students respectively. A selfadministered questionnaire was distributed and nasal swabs were collected. Upon identification, the antibiotic susceptibility of the isolates was examined followed by categorical analysis (Chi-square and Fisher’s exact tests) with factors associated with S. aureus nasal carriage. Twenty one (10%) S. aureus strains were isolated from 209 nasal swab samples. 14 isolates were from pre-clinical students while the remaining seven were from clinical students. There was no significant association between gender, ethnicity, health status, skin infection and students’ exposure to hospital environment with S. aureus nasal carriage (p>0.05). Nineteen (90.5%) isolates were resistant to penicillin and there was also no significant association between penicillin resistant and the students’ groups. One (5.3%) isolate was resistant to erythromycin. There was no methicillin-resistant S. aureus isolated in this study.

Pneumococcal replicative state in relation to its adherence capacity to A549-cell line: A preliminary in vitro analysis

This study was to compare the replication capacity of pneumococcal isolates (serotypes 1, 7F, 19F and 23F) with their adherence pattern to monolayer cells (A549). For standardization purposes, all isolates showed a normal growth curve in both bacteriological (THB + 0.5% yeast extract with and without 2% FBS) and cell culture media (RPMI + 2% FBS). In the former media, a shorter lag phase was observed for isolate serotypes 1 and 7F in presence of serum while in the later; growth yield was lower for all isolates with stationary phase approaching OD600 of 0.01 as compared to 1.0 in bacteriological media. In the replicative analysis at different growth phases of the isolates in cell culture media, growth capacity at 3 h post-incubation was frequently twice as that at 1 h, and that at early-log phase was frequently higher than that at mid-log phase at both post-incubation times. Adherence was frequently the least at early-log phase although the isolates were in the most active state of replication to increase the number of pneumococcal cells to adhere. At mid- and late-log phases, pneumococcal adherence was frequently higher although the replication was reduced. This study marks the potential correlation between pneumococcal growth fitness and adherence capacity whereby the later may not be superior during the early growth phase.

Characterization and antimicrobial activities of two Streptomyces isolates from soil in the periphery of Universiti Putra Malaysia

This study was to assess the identification and antimicrobial activities of two actinomycete isolates. The two isolates designated as B8 and C2, were isolated from a patch of soil in the peripheral area of Universiti Putra Malaysia by streaking on starch casein agar after standard serial dilution procedures. Their antimicrobial activities were first evaluated against eight clinical laboratory strains namely Bacillus sp., Enterococcus sp., Escherichia coli, Klebsiella sp., Pseudomonas sp., Salmonella sp., Staphylococcus aureus, and Staphylococcus epidermidis by perpendicular streak method on Mueller Hinton and Tryptic Soy agar. In both media, a broad-spectrum antibacterial activity was observed for both isolates, with B8 against all the test bacteria and C2 against five of them (Bacillus sp., E. coli, Pseudomonas sp., S. aureus and S. epidermidis). Re-assessment against E. coli ATCC 25922 and S. aureus ATCC 25923 strains by similar method showed antibacterial activities by isolate B8 against both ATTC strains while C2 only against S. aureus ATCC 25923. Streptomyces griseus ATCC 10137 was included in the later experiment and showed antibacterial activity against both ATCC strains. Subsequently, the two isolates were identified by PCR/sequencing techniques and phylogenetic analysis to be Streptomyces species (>93% homology based on 16S rRNA and rpoB genes). Characterization on cultural characteristic and viable count at different temperatures (37ºC and 28ºC), on different microbiological media (AIA, ISP-2, MHA, NA, PDA and TSA), were performed. More morphological features were observed on ISP-2 for both isolates. A higher growth yield was also observed at 28ºC in all media but in comparing that between the two isolates, isolate B8 outnumbered C2 at all experimental conditions. The observed variation in cultural traits and growth yield indicate unique properties between the two antibiotic-producing isolates