Regine Helena Silva dos Fernandes Vieira

Antibacterial effect (in vitro) of Moringa oleifera and Annona muricata against Gram positive and Gram negative bacteria

Antibacterial effects of aqueous and ethanolic extracts of seeds of moringa (Moringa oleifera) and pods of soursop (Annona muricata) in the concentration of 1:5 and 1:10 in volumes 50, 100, 150 and 200 microL were examined against Staphylococcus aureus, Vibrio cholerae, Escherichia coli (isolated from the organism and the aquatic environment) and Salmonella Enteritidis. Antibacterial activity (inhibition halo > 13 mm) against S. aureus, V. cholerae and E. coli isolated from the whiteleg shrimp, Litopenaeus vannmaei, was detected in aqueous and ethanolic extracts of moringa. E. coli isolated from tilapiafish, Oreochromis niloticus, was sensitive to the ethanolic extract of moringa. The aqueous extracts of soursop showed an antibacterial effect against S. aureus and V. cholerae, but the antibacterial activity by the ethanol extracts of this plant was not demonstrated.

In vitro antibacterial effect of aqueous and ethanolic Moringa leaf extracts.

OBJECTIVE To evaluate the antibacterial effect of aqueous and ethanolic moringa leaf extracts (Moringa oleifera) on the growth of gram-positive and negative bacteria. METHODS Paper disks were soaked with 100, 200, 300 and 400 μL of extract at 20 g/180 mL and 10 g/190 mL. All extracts were tested against Escherichia coli (ATCC25922), Staphylococcus aureus (ATCC25923), Vibrio parahaemolyticus, Enterococcus faecalis (ATCC29212), Pseudomonas aeruginosa (ATCC27853), Salmonella enteritidis (IH) and Aeromonas caviae. The susceptibility tests were performed using the modified disk diffusion method. RESULTS The strains E. coli, P. aeruginosa and S. enteritidis (IH) were resistant to all treatments. In general, disks with 400 μL extract were the most efficient against S. aureus, V. parahaemolyticus, E. faecalis and A. caviae. CONCLUSIONS The study indicates a promising potential for aqueous and ethanolic Moringa leaf extracts as alternative treatment of infections caused by the tested strains.

Antimicrobial resistance profile of Vibrio species isolated from marine shrimp farming environments (Litopenaeus vannamei) at Ceara ´ , Brazil

Brazilian shrimp culture industry has a great economic importance mainly to the northeast region. However, the accelerated development of this activity has resulted in the emergency of outbreaks of diseases from farming shrimp, and as a consequence the use of antimicrobial drugs to minimize the potential adverse effect under the shrimp production. The inappropriate use of antibiotics in aquaculture is one of the causes for the high incidence of antimicrobial resistant bacteria isolated from aquatic environments that represent a danger for aquatic organisms and human health. There is little information available on the level of antimicrobial resistance in pathogenic bacteria from shrimp farming environment. Therefore, this study aimed to evaluate the phenotypic resistance profile among Vibrio isolates from hatcheries water samples and from cultivated marine shrimp hepatopancreas (L. vannamei). Antimicrobial susceptibility testing was carried out by a standard disc diffusion method and the minimum inhibitory concentration (MIC) of oxytetracycline (OTC) for resistant Vibrio isolates was determinate by broth dilution method. The results showed a high incidence of resistance to ampicillin (45.2%) and to the tetracycline class (38.7%). Florfenicol and nitrofurantoin were 100% effective against Vibrio isolates. In this study, the OTC-resistant Vibrio spp. showed MIC values of more than 400mg/L and the presence of seawater did not influence the oxytetracycline bioactivity. The occurrence of antimicrobial multiresistance patterns was observed in 29% of Vibrio isolates. Fifty-five percent of multiresistant isolates of Vibrio lost one or more antibiotic resistance phenotype after procedure to curing of resistance plasmids. The oxytetracycline resistance was the phenotype most often lost among plasmid-cured isolates.

Antibacterial activity of GUAVA, Psidium guajava Linnaeus, leaf extracts on diarrhea-causing enteric bacteria isolated from Seabob shrimp, Xiphopenaeus kroyeri (Heller)

O cha de folhas de goiaba Psidium guajava Linnaeus e comumente usado como remedio nas gastrenterites e diarreias infantis por aqueles que nao tem acesso a antibioticos. Esta pesquisa estudou o efeito antibacteriano sobre bacterias causadoras de diarreias, do oleo essencial e do extrato de folhas de goiabeira usando como diluente: metanol, hexano e acetato de etila. Os extratos foram testados sobre Staphylococcus aureus, Salmonella spp. e Escherichia coli. As bacterias testadas foram isoladas de camarao sete-barbas Xiphopenaeus kroyeri (Heller) usando-se como controle cepas padrao, de cada especie. Das bacterias testadas, o melhor efeito inibitorio foi observado sobre Staphylococcus aureus. O extrato de metanol apresentou maior inibicao bacteriana. O oleo essencial mostrou acao inibitoria contra S. aureus e Salmonella spp. Foi demonstrada alguma resistencia das cepas isoladas de camarao aos antibioticos comerciais testados. Estes dados confirmam que remedios a base de folhas de goiabeira podem ser utilizados em casos de diarreia provocada por essas bacterias, quando o uso de antibiotico for restrito. Conclui-se que extratos de folhas de goiaba e o seu oleo essencial sao muito ativos contra S. aureus, tornando-os importantes fontes em potencial de novos compostos antimicrobianos.Guava leaf tea of Psidium guajava Linnaeus is commonly used as a medicine against gastroenteritis and child diarrhea by those who cannot afford or do not have access to antibiotics. This study screened the antimicrobial effect of essential oils and methanol, hexane, ethyl acetate extracts from guava leaves. The extracts were tested against diarrhea-causing bacteria: Staphylococcus aureus, Salmonella spp. and Escherichia coli. Strains that were screened included isolates from seabob shrimp, Xiphopenaeus kroyeri (Heller) and laboratory-type strains. Of the bacteria tested, Staphylococcus aureus strains were most inhibited by the extracts. The methanol extract showed greatest bacterial inhibition. No statistically significant differences were observed between the tested extract concentrations and their effect. The essential oil extract showed inhibitory activity against S. aureus and Salmonella spp. The strains isolated from the shrimp showed some resistance to commercially available antibiotics. These data support the use of guava leaf-made medicines in diarrhea cases where access to commercial antibiotics is restricted. In conclusion, guava leaf extracts and essential oil are very active against S. aureus, thus making up important potential sources of new antimicrobial compounds.

Microbicidal effect of medicinal plant extracts (Psidium guajava Linn. and Carica papaya Linn.) upon bacteria isolated from fish muscle and known to induce diarrhea in children

Out of the twenty-four samples of shrimp and fish muscle used for this study, twelve were collected near a large marine sewer for waste disposal, 3 km off the coast of Fortaleza (Brazil) and used for the isolation of E. coli. Other twelve were collected at the Mucuripe fresh fish market (Fortaleza, Brazil) and used for the isolation of Staphylococcus aureus. Ethanol, water and acetone-diluted extracts of guava and papaya leaf sprouts were tested on the bacteria in order to verify their microbicidal potential. The E. coli strains used in the trials were rated LT positive. The papaya leaf extracts (Carica papaya Linn) showed no microbicidal activity while the guava sprout extracts (Psidium guajava Linn) displayed halos exceeding 13 mm for both species, an effect considered to be inhibitory by the method employed. Guava sprout extracts by 50% diluted ethanol most effectively inhibited E. coli (EPEC), while those in 50% acetone were less effective. It may be concluded that guava sprout extracts constitute a feasible treatment option for diarrhea caused by E. coli or by S. aureus-produced toxins, due to their quick curative action, easy availability in tropical countries and low cost to the consumer.

Microbial contamination of sand from major beaches in Fortaleza, Ceará State, Brazil

The presence of faecal contamination and pathogenic microorganisms in samples of dry and wet sand collected from three major beaches in Fortaleza, Ceara State, Brazil: (Praia do Mucuripe, Praia do Futuro and Praia do Caca e Pesca), during the period of May 1999 to January 2000 was evaluated. Praia do Caca e Pesca had the highest incidence of E. coli in dry sand (56%) followed by Praia do Mucuripe (28%) and Praia do Futuro (16%). In wet sand, results were 48%, 28% and 24% for Praia do Caca e Pesca, Praia do Futuro and Praia do Mucuripe, respectively. Only two samples from Praia do Futuro, one from dry sand and another one from wet sand, were positive for Salmonella. V. parahaemolyticus was isolated from four samples from Praia do Caca e Pesca (two from dry-sand samples and two from wet-sand), one from Praia do Futuro (wet sand), and three and four from Praia do Mucuripe (wet and dry sand, respectively). Yeasts belonged to the Candida genus. Dry-sand samples presented higher yeast contaminations level than wet-sand ones. Praia do Futuro had the highest level of yeast contamination (41%), followed by Praia do Caca e Pesca (33%) and Praia do Mucuripe (26%).

Bacteria of fecal origin in mangrove oysters (Crassostrea rhizophorae) in the Cocó River estuary, Ceará State, Brazil

This study was aimed at evaluating the microbiological quality of mangrove oysters (Crassostrea rhizophorae), collected at a natural oyster bed in the estuary of Coco river (Fortaleza, Ceara, Brazil). MPN values were used for estimating the total (TC) and fecal (FC) coliforms and Enterococcus spp. TC and FC MPN values in the whole muscle and intervalve liquid ranged from 1,600/g and from 1,100/g. No correlation was found between the physico-chemical parameters (temperature, salinity and pH) of the surrounding water and the bacteriological contamination levels found in the tested oysters. The only correlation found was between TC and FC values. Enterococcus spp. strains were isolated and subjected to biochemical tests for species identification. The capacity of those strains for production of a bacteriocin-like inhibitory substance was tested using the Escherichia coli strain ATCC 25922 as a testing organism. Only one, E. faecalis, out of 121 Enterococcus strains tested, presented the inhibitory activity.

Vibrio spp. and Salmonella spp., presence and susceptibility in crabs Ucides cordatus

The presence of Vibrio spp. and Salmonella spp. in crabs marketed at the Bezerra de Menezes Ave., Fortaleza, State of Ceará, Brazil, was assessed between February and May, 2003. The number of individuals sampled in each one of the fifteen weekly samplings ranged between four and eight. Seven strains of Salmonella, from four different samplings, were identified, being five of them identified as serotype S. Senftenberg and two as S. Poona. All strains of Salmonella were sensitive to the tested anti-microbial drugs, with the exception of tetracycline and nalidixic acid, for which an intermediary sensibility was found. The MPNs for Vibrio ranged between 110/g and 110,000/g. Of the forty five Vibrio strains isolated from the crab samples, only 10 were identified up to the species level: two V. alginolyticus and eight V. parahaemolyticus. Bacteria belonging to the Enterobacteriaceae and Pseudomonaceae families were also identified, namely Escherichia coli, Klebsiella pneumoniae, Enterobacter cloacae, Pantoea agglomerans and Pseudomonas aeruginosa. The proper cooking of the animals is recommended in order to avoid problems for the consumers of this crustacean.

Multiple drug resistant Staphylococcus aureus strains isolated from a fish market and from fish handlers

The aim of this study was to investigate the presence of antibiotic resistant Staphylococcus aureus strains in fish stalls and in hands and nasal and oral cavities of fish handlers of the Mucuripe Fish Market, Fortaleza, Ceara, Brazil. All S. aureus isolates were resistant to Ampicillin and 44 % were multi-drug resistant.

Vibrio vulnificus as a health hazard for shrimp consumers

Over the last 30 years, a number of Vibrio species found in the aquatic environment have been indicated as cause of disease in human beings. Vibrio vulnificus is an emergent pathogen, an invasive and lethal marine bacterium related to wound infection and held accountable for gastroenteritis and primary septicemia. It occurs quite frequently in marine organisms, mainly in mollusks. This study aimed at isolating and identifying strains of V. vulnificus based upon the analysis of twenty samples of seabob shrimp, Xiphopenaeus kroyeri (Heller), purchased at the Mucuripe fish market (Fortaleza, Brazil). TCBS agar was used to isolate suspect strains. Seven of twenty-nine strains isolated from six different samples were confirmed as such by means of biochemical evidence and thus submitted to biological assays to determine their virulence. The susceptibility of the V. vulnificus strains to a number of antibiotics was tested. None of the V. vulnificus strains showed signs of virulence during a 24-hour observation period, possibly due to the shedding of the capsules by the cells. As to the results of the antimicrobial susceptibility tests, the seven above-mentioned V. vulnificus strains were found to be sensitive to nitrofurantoin (NT), ciprofloxacin (CIP), gentamicin (GN) and chloramphenicol (CO) and resistant to clindamycin (CI), penicillin (PN) and ampicillin (AP).