Screening of Bioactive Secondary Metabolites from Sea Sponge (Clathria Indica) Against Bacteria Associated with Urinary Tract Infections

The marine sponge Clathria indica, collected from Thondi-Palk Strait region of Tamil Nadu, was studied for bacterial antagonistic activity. Sponge species were identified based on specula morphology. Ethyl Acetate extracts yielded a total of 0.8g, 0.12g, 0.01g, 0.13g and 0.17g from 1.5g of sponge crude extracts respectively. The antagonistic activity of crude extracts against bacterial pathogens showed clear inhibition zones against Pseudomonas sp., Streptococcus sp. and Vibrio sp. The extracted sponge metabolites had growth inhibitory activities against all the three Urinary tract pathogens, Vibrio sp., Pseudomonas sp. and Streptococcus sp. and Bactericidal activities against two Urinary tract pathogens, Vibrio sp. and Pseudomonas sp. The partial isolation of DNA was done by using Gel electrophoresis. On the gel the DNA showed one defined band which had a size of 39.360Kb.Key words: Sea sponge Clathria indica; Vibrio sp.; Pseudomonas sp.; Streptococcus sp.

First-Line Anti-Tuberculosis Drug Susceptibility Patterns of Mycobacterium tuberculosis Complex Strains Responsible for New Cases of Human Pulmonary Tuberculosis in Kisumu County, Western Kenya

Background: Tuberculosis (TB) remains a major cause of morbidity and mortality worldwide, drug-resistant tuberculosis being a major public health problem. The emergence and spread of multidrug resistant (MDR) Mycobacterium tuberculosis complex (MTBC) strains poses significant challenges to disease control. Continued surveillance of drug susceptibility helps determining proper treatment regimen. The effectiveness of a standard anti-tuberculosis (TB) treatment regimen correlates with in vitro drug susceptibility pattern of the infecting tubercle bacilli. The results of the drug susceptibility tests help select a proper treatment regimen or modify treatment regimen for a better management of patients and surveillance and timely control of the spread of the drug resistant TB in the community. Treatment of drug resistant TB is costly, and the outcomes, including survivorship, can be poor. As the result, the drug susceptibility test has become more important than ever. Objective: This study aimed to investigate the patterns of first line anti-tuberculosis drug-susceptibility against Mycobacterium tuberculosis complex isolates from new cases of pulmonary TB patients in Kisumu County, Western Kenya. Method: This was a cross sectional study which included a total of 290 isolates from pulmonary TB patients in JOOTRH and Kisumu County Hospital between February and August 2016. The MTBC isolates identified were M. tuberculosis, M. africanum, and M. bovis. Drug susceptibility test was performed on the 283 M. tuberculosis, 5 M. africanum and 2 M. bovis isolates by BD BACTEC MGIT 960 SIRE and PZA DST system using five first-line anti-TB drugs: Isoniazid, Rifampicin, Streptomycin, Ethambutol and Pyrazinamide. Results: M. tuberculosis was highly sensitive to all the anti-TB drugs; Streptomycin(S) 96.8%, Isoniazid (H) 89.8%, Rifampin(R) 98.2%, Ethambutol (E) 94.4%, Pyrazinamide (PZA) 89.8%. M. bovis TB species was 100% sensitive to all drug except Pyrazinamide where there was 100% resistance. M. africanum varied in its sensitivity to anti-TB drugs; Streptomycin 80%, Isoniazid 60%), Pyrazinamide 4 (80%). Resistance was Streptomycin 20%, Isoniazid 40%, and Pyrazinamide 20%. M. africanum was neither resistant to Rifampin(R) nor Ethambutol (E). A total of 20.8% of M. tuberculosis strains showed resistance to at least one drug tested, while 79.2% were sensitive. 16.3% were resistant to one drug (mono resistance), 2.1% to two drugs (double resistance), 0.7% to three drugs (triple resistance), 0.4% to four drugs (quadruples) and 1.4% to five drugs (pentagon-resistance). Two isolates of M. bovis were resistant to one drug. Two isolates of M. africanum were resistance, one case to one drug and another one case to three drugs. Conclusion: This study showed high level of resistance in M. tuberculosis isolates warranting proper use of anti-TB drugs in Kisumu County. Keywords: Tuberculosis, M. tuberculosis complex, Multi Drug Resistanc

Molecular Identification of Soil Bacteria by 16srDNA Sequence

In this current study, 16S rDNA (genotypic) identification technique is focused on identification of conventionally unidentifiable isolates those are unevaluated in isolated by employing molecular techniques and Bioinformatics in uploading and retrieving isolate gene sequences which are rapid, reliable and accurate in differentiation of various soils isolates. This study is an automaton of 16Sr DNA gene sequence that allows a queue comparison analysis of published sequences deposited in the microbial genome database was used. Polymerase chain reaction (PCR) amplification of 16SrDNA gene using the consensus bacterial primer and separation of the resulting polymer chain reaction amplicon  by cloning, temperature gradient electrophoresis are major ecological techniques that are used in the description of soil bacteria. The isolated gene was cloned using PTZ57r or T cloning Vector amplified using 16SF and 16SR primer transformed in DH5? Cells resulting PCs 16s Plasmid hybrid. The primer 16S F2 obtained from M13 forward primer was used and aligned using BLAST and submitted to EMBL+GENEBANK+DDBJ+ PDB. 99% similarity was observed and later it was analyzed with the existing sequence in ribosomal database project II.  RDP classifier was used for confirmation with 100 % similarity. The bacteria were identified as Burkholderia cenocepacia  when the  sequence was submitted and retrieved via the World Wide Web and new sequence compared with those held in the database  using the basic local alignment tool (BLAST). A segment of 734 out of 736 nucleotide of 16S rDNA gene of Burholderia Cenocepacia is the region of choice for primer construction because of proximity that provides a successful discrimination in strains of Burholderia Cenocepacia in soil. 16S rDNA gene account to 99%  similarity score in molecular typing and identification of bacteria which concerns deposition of sequences into established microbial genomic database Key Words: Burkholderi; Bacterial transformation; Characterization; DNA based technique

Molecular Identity of Mycobacteria Isolates in New Cases of Pulmonary Tuberculosis Patients in Kisumu County, Western Kenya

Background: Pulmonary tuberculosis (TB) remains one of the most challenging diseases to control in the world today and it has become a major global health problem especially in immunocompromised people such as HIV/AIDS. The problem is compounded by the emergence of non-tuberculous mycobacteria (NTM) of which its treatment is not directly analogous to that of MTB. Objective: This study determined the identity of Mycobacteria isolates in new cases of human pulmonary TB patients. Methods: It was a cross-sectional study that involved 316 confirmed new cases of pulmonary TB attending JOOTRH and Kisumu County Hospital. Sputa specimen was cultured in MGIT liquid culture medium. The isolates were identified to species level using GenoType® Mycobacterium CM/AS and MTBC Assay from Hain Lifescience Germany. Results. Of the 316 culture positive isolates, 91.8% were identified as MTBC and 8.2% were NTM species. Of the 290 MTBC, three different species were identified, 97.6% were M. tuberculosis, 1.7% were M. africanum and 0.7% were M. bovis. The Fisher’s exact test was used to assess the associations between patient characteristics and MTBC species identified showed that age category of patients less than 35 years and above 35 years were statistically significant with MTBC species (p=0.020). While sex was not statistically significant with MTBC species (p=0.696). Four different NTM species were identified as 61.5% M. intracellulare, 19.2% M. abscessus, 11.5% M. kansasii and 7.7% M. fortuitum. The Fisher’s exact test done to assess the associations between patient characteristics and NTM species was identified. Age category (p=0.608) and sex (p=0.182) of patients was not statistically significant to NTM species. Conclusion: There is a need for routine speciation among members of the MTBC and NTM as it is an important prerequisite for the proper management of patients with mycobacterial infections. Keywords: Mycobacterium tuberculosis complex, Non tuberculous mycobacteria, Tuberculosi

Isolation, Identification and Characterization of Urinary Tract Infectious Bacteria and the Effect of Different Antibiotics

Introduction: Urinary Tract Infection (UTI) defines a condition in which the urinary tract is infected with a pathogen causing inflammation which is a common, distressing and occasionally life threatening condition. UTI affects people of all  ages and both gender. In all patients with UTI are reported with asymptomatic bacteriuria. Female are more susceptible to  UTIs compared to  male. To ensure appropriate therapy, current knowledge of  the  organisms  that  cause  UTI  and  their  antibiotic  is  susceptibility is mandatory. Methods: This study focused on the frequency of uropathogens and  their antibiotic susceptibility in different gender in Madurai District. Cultural and biochemical characterization  of  uropathogens revealed  the  prevalence  of  both  gram-positive and gram-negative organisms Results: E. coli was the predominant isolate isolated from the urine specimen followed by Pseudomonas aeruginosa, Klebsiella pneumoniae, Staphylococcus aureus, Proteus mirabilis and Enterococcus faecalis. Among the antibiotics tested, chloraphenicol and ciprofloxacin (100%) were found to be effective for empirical treatment of UTI and has covered the majority of urinary pathogens followed by tetracycline,  gentamycin and kanamycin (83%), Ampicillin (67). Streptomycin, Rifampicin and amoxicillin were less effective (50%). Conclusion: Some of the isolates were resistant to penicillin-G, Streptomycin, rifampicin and amoxicillin which are more frequently prescribed and indicates that increased consumption of a particular antibiotic leads to acquisition of resistance by the uropathogens.   Resistance rates among common uropathogens continue to evolve and appear to be increasing too many commonly used antimicrobial agents and a continued surveillance of resistance rates among uropathogens is needed to ensure appropriate recommendations for the treatment of the urinary tract infections. Keywords: Urinary tract infection, Drug resistence, Uropathogens, Biochemical tests

Autoimmune hepatitis type 1 in children : laboratorial features associated with histology and outcome

INTRODUÇÃO: A pesquisa de fatores que possam reconhecer precocemente quais serão os pacientes portadores de HAI com boa evolução ou aqueles que serão resistentes ao tratamento, levaria a um melhor planejamento da terapia. OBJETIVOS: Determinar se fatores laboratoriais pré-tratamento, e o tempo necessário para atingir remissão clínico-laboratorial são preditivos de remissão histológica ou melhora do grau arquitetural na HAI tipo1 em crianças. MÉTODOS: Estudo retrospectivo com 50 crianças portadoras de hepatite autoimune tipo 1 acompanhadas na Unidade de Hepatologia Pediátrica do Instituto da Crianças HC- FMUSP, no período de 1992 a 2012. Realizado revisão de 93 biópsias hepáticas às cegas por um único patologista. RESULTADOS: Foram selecionadas 40 crianças segundo critérios de inclusão. Na biópsia inicial a atividade inflamatória grau 4 e cirrose eram predominantes (31 pacientes-77,5%), sendo que na biópsia de controle, 17 crianças atingiram remissão histológica e 11 melhora do grau arquitetural. Não encontramos valores estatísticos dos fatores laboratoriais ao diagnóstico em relação à remissão histológica. Já em relação à arquitetura, os exames bilirrubina total (p=0,02) e direta (p=0,04) e o tempo de Protrombina (p=0,07) foram relacionados à melhora do grau arquitetural quando apresentavam valores com menor alteração. O tempo necessário para atingir remissão clínico-laboratorial não apresentou correlação com a melhora do padrão inflamatório ou arquitetural. O tratamento foi suspenso em 14 crianças, com taxa de recaída de 50%, em que a maioria ainda apresentava atividade inflamatória na histologia. O tempo de tratamento foi maior nas crianças que evoluíram com remissão sustentada depois de retirada da medicação. CONCLUSÕES: Observou-se em nosso estudo que os fatores laboratoriais ao diagnóstico, independente de seu grau de alteração, não podem predizer quais as crianças que evoluirão para remissão histológica. As crianças com menor grau de alteração de bilirrubina total e direta e tempo de Protrombina, são as que evoluíram com reversão da fibrose hepática. O tempo necessário para atingir a remissão clínico-laboratorial não foi fator preditivo de melhora histológica. Recaída depois da suspensão do tratamento foi associada a presença de atividade inflamatória, e os pacientes em remissão sustentada são os com maior tempo de tratamentoINTRODUCTION: The study of factors that can recognize, earlier, which patients will be carrying autoimmune hepatitis presenting good outcome or those who will be resistant to treatment, would lead to a much better therapy planning. This study aims to: determine whether pretreatment laboratory factors and the time required for achieving clinical and laboratory remission are predictors of histological remission or improvement of the architectural degree in AIH type 1 in children. METHODS: A retrospective study of 50 children with autoimmune hepatitis type 1 accompanied by the Pediatric Hepatology Unit of the Institute of Children HC- FMUSP between 1992 and 2012. A review of 93 liver biopsies was randomly conducted by a single pathologist. RESULTS: Forty children were selected according to criteria of inclusion. In the initial biopsy the inflammatory activity level 4 and cirrhosis were predominant (31 patients-77, 5%), in the control biopsy 17 patients achieved histological remission and 11 presented improvement in the architectural degree. We found no statistical values of the diagnostic laboratory factors in relation to histological remission. Regarding the architecture, the exams total bilirubin (p = 0.02) and direct bilirubin (p = 0.04) and prothrombin time (p = 0.07) were related with the improvement of the architectural level when presented the values with lower changes. The time required for achieving clinical and laboratory remission presented no correlation with the improvement of the inflammatory or architectural pattern. The treatment was suspended in 14 children with relapse rate of 50%, in most of them the inflammatory activity still present on histology. The length of treatment was longer in children who developed sustained remission after the withdrawal of medication. CONCLUSIONS: We observed in our study that the laboratory factors for the diagnosis, regardless their degree of alteration, can not predict which children will progress to histological remission. The children who presented lower degrees of change in total and direct bilirubin and prothrombin time are those who have progressed to reversal of liver fibrosis. The time required for achieving clinical and laboratory remission was not a predictive factor for histological improvement. Relapse after discontinuation of treatment was associated to the presence of inflammatory activity, and patients with sustained remission are the ones who had longer period of treatmen