Jurnal Teknologi Laboratorium
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Total Lymfosit Count (TLC) with CD4 in HIV/AIDS Patients at Kupang
Get PDFHuman Immunodeficiency Virus (HIV) is a retrovirus originating from the retroviridae family of the genus lentivirus that infects and damages cells that have a molecule Cluster of Differentiation 4 (CD4), especially T lymphocytes that have receptors with high affinity for HIV. Total lymphocyte count / TLC has been proposed as an alternative guide to CD4 in limited health facilities. This study aims to determine the correlation between Total Lymphocyte Count (TLC) and CD4 in HIV/AIDS patients in the W.Z. Johannes Kupang hospital. The type of this research was observational analytic with a cross-sectional design. The study was conducted on 121 samples of patients who performed CD4 examination and Total Lymphocytic Count (TLC) in the laboratory of W.Z.Johannes Kupang Hospital. The Spearman correlation test shows a significance value of 0,000 with a Spearman correlation value of 0.799. Based on the results of this study it can be concluded that there is a significant correlation between Total Lymphocyte Count and CD4 and shows the direction of positive correlation with a very strong relationship, where the increase in the number of Total Lymphocyte Count is in line with the increase in CD4 counts
Purification and characterization of recombinant Streptokinase expressed in E.coli from Streptococcus equisimilis with N-terminal methionine
No full textStreptokinase is a extracellular enzyme which is extracted from strains of beta Hemolytic streptococcus. The enzyme is a non-protease plasminogen activator that activates plasminogen to plasmin and degrades fibrin clot through its specific lysine binding site which is used in thrombolytic therapy. Purification of streptokinase produced from S.equisimilis in E.coli with N-terminal methionine was carried out in 3 Chromatography purification steps, 1) CM-Sepharose-FF at pH 4.2 followed by concentration and dialysis over night with Tris-HCl pH 8.0. Partially purified dialyzed enzyme sample was loaded on to 2) DEAE-Sepharose-FF column. The Purified fractions of DEAE column were pooled and applied on to Sephadex G-100 column. Enzyme purity was confirmed by SDS-PAGE and RP-HPLC.Its biological activity is determined by specific streptokinase assay and characterised the enzyme by Peptide mapping, MALDI-TOF, Isoelectric-focusing and RP-HPLC. The isoelectric point (pI) of streptokinase is around 4.98.The results of characterization shows that it contains two forms (Isomers) of streptokinase expressed in E. coli which was analyzed by RP-HPLC and chromogenic assay. The variation is formed by isomer-1 in which 85% of Streptokinase expressed without methionine (85000IU/mg) and Isomer-2 in which 15% of streptokinase expressed with methionine (nil activity) in E. coli. This phenomenon shows that the presence and absence of methionine in isomers of streptokinase varying the catalytic activity of the enzyme
Potential of Lactobacillus casei shirota’s strain against the biofilm-forming of Salmonella Spp
Get PDFBiofilm of Salmonella spp. is formed through the expression of biofilm genes associated with proteins (bapA) regulated by curli synthesis genes (csg) which carry out adhesion, colonization, maturation, and dispersion on the surface of the intestinal epithelium. This study aimed to determine the antibiofilm activity of Lactobacillus casei Shirota’S strain (LcS) as an inhibitor of Salmonella spp. biofilm formation in vitro. This research was a true experimental study using Microtiter Plate 96 wells Biofilm Assay method. The sample used was the suspension of Salmonella spp. The treatment was in the form of adding a LcS suspension with a concentration series of 10-1;10-2; 10-3;10-4; and 10-5. Biofilm measurements were carried out using a microplate reader and obtained quantitative data in the form of Optical Density at a wavelength of 595nm. The results of this study showed that LcS suspension has antibiofilm activity ranging from 10-5 concentrations with a percentage of 36.58% (p<0.05). The results of exometabolism LcS can reduce Salmonella growth. Exopolysaccharide (EPS) and sortase-dependent proteins (SrtA) of LcS form barriers as competitive adhesion in inhibiting pathogenic biofilm formation.Biofilm of Salmonella spp. is formed through the expression of biofilm genes associated with proteins (bapA) regulated by curli synthesis genes (csg) which carry out adhesion, colonization, maturation, and dispersion on the surface of the intestinal epithelium. This study aimed to determine the antibiofilm activity of Lactobacillus casei Shirota’S strain (LcS) as an inhibitor of Salmonella spp. biofilm formation in vitro. This research was a true experimental study using Microtiter Plate 96 wells Biofilm Assay method. The sample used was the suspension of Salmonella spp. The treatment was in the form of adding a LcS suspension with a concentration series of 10-1;10-2; 10-3;10-4; and 10-5. Biofilm measurements were carried out using a microplate reader and obtained quantitative data in the form of Optical Density at a wavelength of 595nm. The results of this study showed that LcS suspension has antibiofilm activity ranging from 10-5 concentrations with a percentage of 36.58% (p<0.05). The results of exometabolism LcS can reduce Salmonella growth. Exopolysaccharide (EPS) and sortase-dependent proteins (SrtA) of LcS form barriers as competitive adhesion in inhibiting pathogenic biofilm formation
The Presence of Methanol In Alcoholic Beverages Analyzed Using Qualitative Method
No full textAn alcoholic beverage contains ethyl alcohol or ethanol (C2H5OH)that is produced from fermentation or distillation of sugars. In many cases, either producers or consumers add ethanol with denaturated alcohol containing methanol (CH3OH) as an additive. Methanol can cause blindness and induce comas, and it is deadly in high doses. This study aimed at investigating the presence of methanol or methyl alcohol in alcoholic beverages sold in Palembang, Indonesia. Seventeen samples collected from small shops and supermarkets were taken by accidental sampling. A chromotropic acid method was used to examine the presence of methanol. The results showed that there were 18% of the samples was positive, and 82% was negative. Based on alcohol content, the research showed that all (100%) samples of group A were negative; 33% of group B was positive, and 33% of group C was positive. The study indicated that methanol was still present in alcoholic drinks sold in markets. The government should inform the society that denatured alcohol contains methanol and, therefore, should not be feasible to consume
Optimasi Waktu Produksi Metabolit Sekunder dan Skrining Aktivitas Antibakteri Isolat Actinomycetes Rizosfer Tanaman Tin (Ficus carica)
No full textSome Actinomycetes isolates of a tin plant (Ficus carica L.) have been obtained, namely T24M, T18, T19, T24, T25, T34, T37, T41, and T43. The aim of this study was to optimize the production of secondary metabolites (antibiotics) and screening antibacterial activity against Methicillin-Resistant Staphylococcus aureus (MRSA) from the Actinomycetes isolate of the tin rhizosphere. The study was performed with test an activity of the culture fluid from Actinomycetes isolate against MRSA by the well method. The result of optimization secondary metabolite production showed that the second day was the best incubation time to harvest antibiotics. The results showed that bacterial isolates of T24M produced antibiotics that could inhibit MRSA growth
Potensi Ekstrak Daun Carica Pubescens Sebagai Alternatif Antidiare Bakteri Vibrio cholerae dan Shigella dysentriae
No full textAcute diarrhea is one of the main causes of morbidity and mortality. A people are starting to choose traditional medicines for alternative therapy. Traditional medicines or herbal medicines are considered safer and do not have side effects such as chemical drugs. The purpose of this study was to determine the anti-diarrhea effect of Carica pubescens leaf extract on the bacteria Vibrio cholerae and Shigella dysentriae. This study was conducted by testing the activity of Vibrio cholerae and Shigella dysentriae bacteria on Carica pubescens leaf extract with a well method, which results can be seen from the formation of inhibitory zones. The data obtained were processed using Two Way ANOVA test statistics. The results showed that the leaves extract of Carica Pubescens concentration of 100% had the best therapeutic effect because it had the greatest inhibitory power on the bacteria Vibrio cholerae and Shigella dysentriae.Diare akut merupakan salah satu penyebab utama morbiditas dan mortalitas. Masyarakat mulai memilih obat tradisional atau obat herbal sebagai pengobatan alternatif. Obat tradisional atau obat herbal dinilai lebih aman dan tidak memberikan efek samping seperti obat kimia. Tujuan dari penelitian ini adalah untuk mengetahui efek anti diare ekstrak daun Carica pubescens terhadap bakteri Vibrio cholerae dan Shigella dysentriae. Penelitian ini dilakukan dengan menguji aktivitas bakteri Vibrio cholerae dan Shigella dysentriae terhadap ekstrak daun Carica pubescens dengan metode sumuran yang hasilnya dapat dilihat dari terbentuknya zona hambat. Data yang diperoleh diolah menggunakan statistik uji Two Way ANOVA. Hasil penelitian menunjukkan ekstrak daun Caricap pubescens konsentrasi 100% mempunyai efek terapi terbaik karena mempunyai daya hambat terbesar terhadap bakteri Vibrio cholerae maupun Shigella dysentriae
Potensi Antibakteri Isolat Actinomycetes terhadap Aktivitas Proteolitik dan Amilolitik Escherichia Coli ATTC 25922
No full textOccurs E. coli resistance to class 3 cephalosporin class antibiotics and fluoroquinolone groups. The antibiotic resistance that occurs has narrowed the choice of therapy. This study aims to determine the effect of Actinomycetes isolates on proteolytic and amylolytic enzymes E. coli ATCC 25922. This research is experimental research, qualitative tests of protease and amylase enzymes from E. coli ATCC 25922 shown by clear zones around the growing colonies. The result of ANOVA One-Way test showed a significant difference in the width of clear zone, colony zone and PER and AER score with p-value < 0,05. This indicates that Actinomycetes isolates contain compounds that can act as inhibitors of protease and amylase enzymes from E.coli ATCC 25922. It is hoped that there will be research about the identification of Actinomycetes species isolates in Bogor Botanical Garden so that later can be cultivated and produced as an antibiotics alternative
Aktivitas Antibakteri Kulit Buah Karika Dieng terhadap Shigella flexneri dan Escherichia coli
No full textKarika is an endemic plant of Dieng Plateau. The leaf of Karika has been studied to have antibacterial activity against bacteria that cause diarrhea, while the peels are removed or used as a mixture of animal feed. This study aims to determine the antibacterial activity of Karika peels against bacteria that cause poisoning, i.e Shigella flexneri, and Escherichia coli. The sample fractionated to obtain the fraction of n-hexane and ethyl acetate. The fractions were tested for their antibacterial activity against Shigella flexneri and Escherichia coli by the cup-plate method. The results showed that the ethyl acetate fraction (ethyl acetate 50%) had the highest inhibition on Shigella flexneri and Escherichia coli compared with another concentration. Based on the study can be concluded that the peels of Karikahas antibacterial activity against Shigella flexneri and Escherichia coli.Karika merupakan tanaman endemik Dataran Tinggi Dieng. Daun Karika telah diteliti memiliki aktivitas antibakteri terhadap bakteri penyebab diare, sedangkan kulit buah dibuang atau dijadikan campuran pakan ternak. Penelitian ini bertujuan untuk mengetahui aktivitas antibakteri kulit buah Karika terhadap bakteri penyebab keracunan, Shigella flexneri dan Escherichia coli. Sampel difraksinasi untuk memperoleh fraksi n-heksan dan etil asetat. Fraksi diuji aktivitas antibakteri nya terhadap bakteri Shigella flexneri dan Escherichia coli dengan metode sumuran. Hasil penelitian menunjukkan bahwa fraksi etil asetat dengan konsentrasi 50% memiliki daya hambat tertinggi terhadap Shigella flexneri dan Escherichia coli dibandingkan dengan konsentrasi lainnya. Berdasarkan hasil penelitian disimpulkan bahwa kulit buah Karika memiliki aktivitas antibakteri terhadap bakteri Shigella flexneri dan Escherichia coli
Pengembangan Prekultur Oxgall sebagai Sampel Klinis untuk Deteksi Salmonella typhi dengan Metode Real-time PCR
No full textTyphoid fever is a significant public health burden in low-income countries caused by Salmonella enterica serotype typhi (S.typhi). Clinical manifestations of typhoid fever are varied and non-specific, making the diagnosis difficult. Using oxgall for pre-incubation as a selective culture medium before amplification of Real-time PCR (RT-PCR) in wholeblood produces a fast and sensitive diagnostic. The purpose of this study was to know the performance oxgall-precultured Real-time PCR for detection of Salmonella sp.. Prior to the sample process, spike method optimization was performed to find out that the reagents were well used for clinical specimens. In the sample process, blood samples from 30 Widal-positive patients were collected for this study . Venous blood samples from typhoid fever patients were taken on the day of diagnosis; 5 ml for blood culture, and 5 ml for RT-PCR. The bacteria were grown in oxgall 10% (standard microbiological laboratory clinics) and incubated for 6 hours (37° C) before bacterial DNA was isolated for RT-PCR detection. The results showed that reagen of RT-PCR is good used for a clinical sample and a blood culture was better than RT-PCR using oxgall (positive blood culture results over 24 hours). This suggests that there is a need for further research on the duration of incubation and oxgall concentrations in RT-PCR and the selection of clinical samples.Demam tifoid merupakan beban kesehatan masyarakat yang signifikan di negara berpenghasilan rendah yang disebabkan oleh Salmonella enterica serotype typhi (S.typhi). Manifestasi klinis demam tifoid bervariasi dan tidak spesifik, sehingga membuat diagnosis menjadi sulit. Dengan menggunakan oxgall untuk pra-inkubasi sebagai media kultur selektif sebelum amplifikasi Real-time PCR (RT-PCR) pada wholeblood menghasilkan diagnostic yang cepat dan sensitif. Tujuan dari penelitian ini adalah untuk mengetahui kinerja oxgall sebelum amplifikasi RT-PCR untuk deteksi Salmonella sp . Sebelum proses pengambilan sampel, dilakukan optimasi spike sampel untuk mengetahui bahwa reagen yang digunakan baik untuk spesimen klinis. Dalam proses sampel, sampel wholeblood diambil dari 30 pasien dengan positif Widal tes. Sampel darah vena dari pasien demam tifoid diambil pada hari diagnosis; 5 ml untuk kultur darah, dan 5 ml untuk RT-PCR. Bakteri ditanam di oxgall 10% (standar mikrobiologi laboratorium klinik) dan diinkubasi selama 6 jam (37 ° C) sebelum DNA bakteri diisolasi untuk deteksi RT-PCR. Hasil penelitian menunjukkan bahwa reagen RT-PCR baik digunakan untuk sampel klinis dan kultur darah lebih baik daripada RT-PCR dengan menggunakan oxgall (hasil kultur darah positif lebih dari 24 jam). Hal ini menunjukkan bahwa harus ada penelitian lebih lanjut mengenai durasi inkubasi dan konsentrasi oxgall pada RT-PCR dan pemilihan sampel klinis.
 
Antibiotic Resistant and Plasmid Conjugative Study of Salmonella typhi
No full textThe emergence of multi-drug resistant (MDR) bacteria has endangered the efficacy of antibiotics treatment of pathogenic bacteria worldwide. The aim of this research was to investigate the incidence of Salmonella enterica serovar Typhi in Duhok city, Iraq. Specimens of blood and stool were recruited from 267 patients. S. Typhi isolates were diagnosed depending on morphology, biochemical and serological tests. S. Typhi isolates were tested for their antibiotic resistance. Multi-drug resistant S. Typhi isolates were conjugated with E. coli HB101. The plasmid profile of transconjugants was investigated. 15/267 (5.6%) S. Typhi isolates were identified. Based on their biochemical tests, S. Typhi isolates were categorized into two biotypes (I, 26.66% and II, 73.33%). Four resistance patterns were observed. The resistant pattern to ampicillin and tetracycline was the higher (46.6%). Conjugation experiment showed that all antibiotic markers were transferred from S. Typhi to E. coli HB101 with a conjugation frequency of (0.38×10-5). 13.3% of the S. Typhi isolates were multi-drug-resistant resistant and had two small plasmids. Transconjugants E. coli acquired the resistance from the multi-drug resistant S. Typhi. Antibiotics treatment of the pathogens could be hindered by the constant rise of multi-drug-resistant. Further studies are needed to study the mobile genetic elements and their contribution to antibiotics resistance