Study of the antibacterial activity in the gas phase of a chemical formulation for household waste management.

UNLABELLED: The aim of this study was to formulate a product (microbicide mixture) that could slow down the bacterial proliferation during the storage of household waste. We used harmless and natural components, known for their antimicrobial properties, in the liquid phase at direct contact with the microbes. The antimicrobial activity of the microbicide mixture formulated was evaluated over a range of concentration in two types of tests, in the liquid and in the gas phase. Once the efficacy of antimicrobial agent in the liquid phase in direct contact with the microbe (Escherichia coli) was confirmed, we adopted a new approach to evaluate the effect of the vapour phase both on the microbes' growth and on its duration. Here, we show that the perfect combination that gives rise to an antimicrobial mixture useful to control microbial growth (Staphylococcus aureus, Escherichia coli, Debaryomyces hansenii or Penicillium citrinum) up to 4 weeks is the one between more volatile agents (2-propanol and limonene) and a less volatile agent (cinnamaldehyde). The pleasant smell as well as the synergic antibacterial and antifungal function of the natural components of this mixture makes it attractive in domestic waste management. SIGNIFICANCE AND IMPACT OF THE STUDY: The novelty of this work is two-fold: on the one hand, to test various antimicrobial components of different volatility in a single microbicide mixture, and on the other, to study antimicrobial activity in the gas phase, other than the liquid phase. While previous authors tested the components individually as antimicrobial agents in the liquid phase at direct contact with the microbes, we tested them altogether as a mixture both in the liquid and in gas phase. The aim of this study was to disinfect small environments, such as garbage containers, by favouring the diffusion of the vapour phase to avoid the growth of microbes. This study proposes a new approach in the management and storage of household waste by inhibiting bacterial proliferation in the garbage can.

Isolation of Pantoea ananatis from sugarcane and characterization of its potential for plant growth promotion.

Each year, approximately 170 million metric tons of chemical fertilizer are consumed by global agriculture. Furthermore, some chemical fertilizers contain toxic by-products and their long-term use may contaminate groundwater, lakes, and rivers. The use of plant growth-promoting bacteria may be a cost-effective strategy for partially replacing conventional chemical fertilizers, and may become an integrated plant nutrient solution for sustainable crop production. The main direct bacteria-activated mechanisms of plant growth promotion are based on improvement of nutrient acquisition, siderophore biosynthesis, nitrogen fixation, and hormonal stimulation. The aim of this study was to isolate and identify bacteria with growth-promoting activities from sugarcane. We extracted the bacterial isolate SCB4789F-1 from sugarcane leaves and characterized it with regard to its profile of growth-promoting activities, including its ability to colonize Arabidopsis thaliana. Based on its biochemical characteristics and 16S rDNA sequence analysis, this isolate was identified as Pantoea ananatis. The bacteria were efficient at phosphate and zinc solubilization, and production of siderophores and indole-3-acetic acid in vitro. The isolate was characterized by Gram staining, resistance to antibiotics, and use of carbon sources. This is the first report on zinc solubilization in vitro by this bacterium, and on plant growth promotion following its inoculation into A. thaliana. The beneficial effects to plants of this bacterium justify future analysis of inoculation of economically relevant crops.

Identification and antimicrobial resistance of microflora colonizing feral pig (Sus scrofa) of Brazilian Pantanal.

Antimicrobial resistance of bacteria is a worldwide problem affecting wild life by living with resistant bacteria in the environment. This study presents a discussion of outside factors environment on microflora of feral pigs (Sus scrofa) from Brazilian Pantanal. Animals had samples collected from six different body sites coming from two separated geographic areas, Nhecolandia and Rio Negro regions. With routine biochemical tests and commercial kits 516 bacteria were identified, with 240 Gram-positive, predominantly staphylococci (36) and enterococci (186) strains. Among Gram-negative (GN) bacteria the predominant specimens of Enterobacteriaceae (247) mainly represented by Serratia spp. (105), Escherichia coli (50), and Enterobacter spp. (40) and specimens not identified (7). Antimicrobial susceptibility was tested against 17 drugs by agar diffusion method. Staphylococci were negative to production of enterotoxins and TSST-1, with all strains sensitive towards four drugs and highest resistance toward ampicillin (17%). Enterococci presented the highest sensitivity against vancomycin (98%), ampicillin (94%) and tetracycline (90%), and highest resistance pattern toward oxacillin (99%), clindamycin (83%), and cotrimoxazole (54%). In GN the highest resistance was observed with Serratia marcescens against CFL (98%), AMC (66%) and AMP (60%) and all drugs was most effective against E. coli SUT, TET (100%), AMP, TOB (98%), GEN, CLO (95%), CFO, CIP (93%). The results show a new profile of oxacillin-resistant enterococci from Brazilian feral pigs and suggest a limited residue and spreading of antimicrobials in the environment, possibly because of low anthropogenic impact reflected by the drug susceptibility profile of bacteria isolated.

Disinfection of roof harvested rainwater inoculated with E. coli and Enterococcus and post-treatment bacterial regrowth: Conventional vs solar driven advanced oxidation processes.

Solar driven advanced oxidation processes (AOPs) (an alternative solar photo Fenton like process (SPF), sunlight/H2O2 (SHP) and sunlight/chlorine (SCL)) and respective dark conditions, were compared for the first time to conventional (chlorination and UV-C radiation) disinfection processes, in the inactivation of E. coli and Entero strains inoculated in real roof-harvested rainwater (RHRW), to evaluate their possible safe use for crop irrigation. In this regard, bacterial regrowth was also evaluated 6, 12, 24 and 48 h after disinfection treatment. The SPF, using iminodisuccinic acid (IDS)-Cu complex as catalyst, was optimized (H2O2/IDS-Cu 55/1 best molar ratio) under mild conditions (spontaneous pH) and sunlight. The faster inactivation kinetics were observed for the SCL process (k = 1.473 min-1, t1/2 = 0.47 min for E. coli and k = 1.193 min-1, t1/2 = 0.57 min for Entero), while the most effective processes in controlling bacterial regrowth were SPF and SCL. Although UV-C radiation (0-1.3 × 104 µW s cm-2 dose range) was the second faster disinfection process (k = 1.242 min-1, t1/2 = 0.55 min for E. coli and k = 1.150 min-1, t1/2 = 0.60 min for Entero), it was the less effective process in controlling bacterial regrowth (>10 CFU 100 mL-1 already after 6 h post-treatment incubation). According to the bacterial inactivation and regrowth tests carried out in this work, SPF and SCL are interesting options for RHRW disinfection, in case of effluent use for crop irrigation.

New approach for evaluating the public health risk of living near a polluted river.

Chemical, physical and microbial analyses were conducted in the Sarno River basin to obtain a comprehensive description of the overall quality of the water bodies. The collection period lasted 12 months, between 2005 and 2006, with high frequency of sampling and analysis. More than 6,000 analytical determinations were performed on samples collected at six sampling points along the Sarno River and two points each on tributaries Solofrana and Cavaiola. The results indicated the presence of inorganic contaminants, which, in most cases, were below the Italian State water quality thresholds. The organic contamination showed an increasing trend, with respect to previous determinations, thus demonstrating the major contribution of untreated urban wastewater to the overall pollution of the river. Moreover, this study was designed to explore the correlation between the presence of microbial indicators of fecal contamination in Sarno River and their presumable presence in the aerosol surrounding the river, thus pointing to the possible environmental hazard associated with the presence of pathogens in the air.

Two-years surveillance of fungal contamination in three hospital departments in Campania region.

A two-years (2003-2005) fungal environmental surveillance was carried out in three departments of a hospital in Campania region (Surgery, Intensive Care Unit, Obstetrics and Gynaecology). Four operating theatres rooms and their relative areas of service and support, 4 patient rooms of intensive care unit, 1 delivery room, 1 labour room and 1 nursery of Obstetrics and Gynaecology were checked. A total of 12,120 surfaces and 2,904 air samples were collected in 24 monthly determinations. A seasonal variation in the fungal development was observed, in particular the lowest level of air and surface fungi contamination was found in winter and autumn whereas it was higher in spring and summer. In this study 30 fungal species were identified and, among these, the most frequent specie isolated was the Aspergillus spp. The results show an air contamination, expressed in percentage of positive determinations for Aspergillus spp, and the other fungi in the following percentages: Obstetrics and Gynaecology (25% and 33,3%); Intensive Care Unit (17% and 25%); Surgery (12.5% and 21%). For surfaces contamination it was found: Obstetrics and Gynaecology (67% and 75%); Intensive Care Unit (63% and 71%); Surgery (58.3% and 67%). This study shows that in the departments observed environmental fungi contamination is always present and therefore it would be necessary to apply environmental surveillance procedure and monitor the effectiveness.

Surveillance of nosocomial infections: a preliminary study on hand hygiene compliance of healthcare workers.

The observance of hand hygiene compliance is important to reduce cross-infection by micro-organisms. The aim of this preliminary study was to evaluate the level of hand hygiene in healthcare workers from different departments, with particular emphasis on transient flora. The study was conducted in three departments (Surgery, Intensive Care Unit, Obstetrics and Gynecology) of a hospital in Campania, southern Italy. Over a six-month period, 50 healthcare workers were randomly tested. Imprints of palms and fingertips were taken monthly during the morning shift. The number of colonies per plate was counted and transient pathogens were identified. Risk factors for hand contamination were determined. Total flora was found in the following CFU means per palm and per five fingertips (95% CI): Obstetrics and Gynecology [palms 130 CFUs (95% CI 85-180); fingertips 125 CFUs (95% CI 92-160)]; ICU [palms 80 CFUs (95% CI 58-99); fingertips 62 CFUs (95% CI 45-82)]; Surgery [palms 75 CFUs (95% CI 41-120); fingertips 70 CFUs (95% CI 52-90)] Transient flora was found on 39% of healthcare workers' hands. The only factor associated with hand contamination by transient flora was the absence of gloving during healthcare procedure (P = 0.02).

RNAIII inhibiting peptide (RIP) inhibits agr-regulated toxin production.

Staphylococcal enterotoxins (SEs) are emetic toxins that cause food poisoning. SEs also function as powerful pyrogenic toxin superantigens that stimulate non-specific T-cell proliferation. Together with the hemolysins, SEs have been largely implicated as virulence factors in multiple infection models. Recent biochemical and genetic analyses have demonstrated that production of some of these toxins is partially regulated by quorum sensing mechanisms where proteins and peptides activate the accessory gene regulator (agr). Because toxin production is central to bacterial pathogenesis, therapeutic strategies alternative to antibiotics, and based on rational interference of the quorum sensing systems involved, are currently being developed. This approach would lead to repression of toxin production and, thus, to disease prevention. Here we provide evidence to conclude that synthetic analogs of the RNAIII inhibiting peptide (RIP) and antibodies to its target molecule TRAP function in vitro as efficient suppressors of agr-regulated exotoxin production by Staphylococcus aureus.

Prevention of diseases caused by Staphylococcus aureus using the peptide RIP.

Staphylococcus aureus causes many diseases including cellulitis, keratitis, osteomyelitis, septic arthritis and mastitis. The heptapeptide RIP has been shown to prevent cellulitis in mice, which was induced by S. aureus strain Smith diffuse. Here we show that RIP can also significantly reduce the overall pathology and delay the onset of disease symptoms in several other models of S. aureus infections, including: keratitis (tested in rabbits against S. aureus 8325-4), osteomyelitis (tested in rabbits against S. aureus MS), mastitis (tested in cows against S. aureus Newbould 305, AE-1, and environmental infections) and septic arthritis (tested in mice against S. aureus LS-1). These findings substantiate that RIP is not strain specific in its inhibitory activity and that RIP is an effective inhibitor of bacterial pathology at multiple body sites following diverse routes and doses of administration. These findings strongly evidence the potential value of RIP as a chemotherapeutic agent.

Serological and molecular investigation of the prevalence of Aujeszky's disease in feral swine (Sus scrofa) in the subregions of the Pantanal wetland, Brazil.

The feral swine (FS) originated from the domestic pig and is present throughout the Brazilian wetland plain (the Pantanal). Aujeszky's disease (AD) was first serologically confirmed in the state of Mato Grosso do Sul (MS) in 2001; however, there was no viral confirmation. The aim of this study was to investigate antibodies against-SuHV-1 in the sera of feral swine in the studied areas, detect SuHV-1 through PCR and classify the viral genome. Among the 218 animals sampled, 186 were analyzed by ELISA, resulting in 88 (47.3%) reactive samples. In the serum neutralization test (SN), 57/179 (31.8%) samples presented antibodies against the AD virus (SuHV-1). By nested PCR, 104 DNA samples were extracted for analysis and confirmed with amplification of a fragment of glycoprotein B (gB) in five samples. The SuHV-1 was detected in 12 samples by using primers for glycoprotein E (gE) and viral genome was classified as Type I by ul44 partial sequencing. The amplification of SuHV-1 glycoprotein fragments in the fetuses of seropositive sows indicate that the vertical transmission contribute to maintain SuHV-1 in a free-living feral swine population. The origin of AD in the feral swine populations of the Pantanal is unknown, however, the determination of viral latency, the vertical transmission of the antigen by the amplification of SuHV-1 glycoprotein fragments in the fetuses of seropositive sows and genome typing contribute to the elucidation of the epidemiology of this disease in the wetlands of MS, Brazil.

Identification and antimicrobial resistance of microflora colonizing feral pig (Sus scrofa) of Brazilian Pantanal

Antimicrobial resistance of bacteria is a worldwide problem affecting wild life by living with resistant bacteria in the environment. This study presents a discussion of outside factors environment on microflora of feral pigs (Sus scrofa) from Brazilian Pantanal. Animals had samples collected from six different body sites coming from two separated geographic areas, Nhecolandia and Rio Negro regions. With routine biochemical tests and commercial kits 516 bacteria were identified, with 240 Gram-positive, predominantly staphylococci (36) and enterococci (186) strains. Among Gram-negative (GN) bacteria the predominant specimens of Enterobacteriaceae (247) mainly represented by Serratia spp. (105), Escherichia coli (50), and Enterobacter spp. (40) and specimens not identified (7). Antimicrobial susceptibility was tested against 17 drugs by agar diffusion method. Staphylococci were negative to production of enterotoxins and TSST-1, with all strains sensitive towards four drugs and highest resistance toward ampicillin (17 percent). Enterococci presented the highest sensitivity against vancomycin (98 percent), ampicillin (94 percent) and tetracycline (90 percent), and highest resistance pattern toward oxacillin (99 percent), clindamycin (83 percent), and cotrimoxazole (54 percent). In GN the highest resistance was observed with Serratia marcescens against CFL (98 percent), AMC (66 percent) and AMP (60 percent) and all drugs was most effective against E. coli SUT, TET (100 percent), AMP, TOB (98 percent), GEN, CLO (95 percent), CFO, CIP (93 percent). The results show a new profile of oxacillin-resistant enterococci from Brazilian feral pigs and suggest a limited residue and spreading of antimicrobials in the environment, possibly because of low anthropogenic impact reflected by the drug susceptibility profile of bacteria isolated.

Diagnóstico microbiológico e histopatológico de mortalidade em avestruzes (Struthio camelus) / Microbiological and histological diagnosis in mortality of ostrich (Struthio camelus)

Several young ostrich, including nestlings, with lassitude and inappetence followed by death or victim of sudden death were immediately brought to diagnosis at an Animal Health Laboratory. At necropsy, animals presented hemorrhage and altered content of the vitelline sac, and necrotic foci in the small intestine; one animal showed necrotic pleuropneumonia with psammomatosus bodies in the lung parenchyma. The cultures from different samples revealed Escherichia coli, Pseudomonas aeruginosa, Klebsiella pneumoniae, Serratia marcescens, Enterobacter aglomerans, and Pseudomonas mendocina. It was suggested one case of septicemia in an animal with exclusive growth of K. pneumoniae isolated from samples of small intestine, lung, and liver.