Prevalence and resistance to antibiotics for Aeromonas species from retail fish in Malaysia.

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Radu, Son; Ahmad, Noorlis; Ling, Foo Hooi; Reezal, Abdul

2003-03-25

A total of 87 market fish samples representing five types of fish were evaluated for the presence of Aeromonas spp. Of the samples examined, 69%, 55%, 11.5% and 2.3% harbored Aeromonas spp., A. veronii biovar sobria, A. hydrophila and A. caviae, respectively. The 60 isolated Aeromonas spp. strains were further examined for hemolytic activity, resistance to antimicrobial agents and presence of plasmids. Hemolytic activity varied widely among the isolated strains. Though all the isolates demonstrated resistance to three or more of the antibiotics tested, all were susceptible to ceptazidime. Thirty-four (56.7%) of the sixty isolates harbored plasmids, with sizes ranging from 2.3 to 15.7 kb. These results indicate that hemolytic, multiple antibiotic resistant and genetically diverse aeromonads are easily recovered from fish in this region.

Effectiveness of radiation processing in elimination of Aeromonas from food

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Nagar, Vandan; Bandekar, Jayant R.

2011-08-01

Genus Aeromonas has emerged as an important human pathogen because it causes a variety of diseases including gastroenteritis and extra-intestinal infections. Contaminated water, sprouts, vegetables, seafood and food of animal origin have been considered to be the important sources of Aeromonas infection. In the present study, radiation sensitivity of indigenous strains of Aeromonas spp. from different food samples was evaluated. The decimal reduction dose (D10) values of different Aeromonas isolates in saline at 0-4 °C were in the range of 0.031-0.046 kGy. The mixed sprouts, chicken and fish samples were inoculated with a cocktail of five most resistant isolates (A. salmonicida Y567, A. caviae A85, A. jandaei A514A, A. hydrophila CECT 839T and A. veronii Y47) and exposed to γ radiation to study the effectiveness of radiation treatment in elimination of Aeromonas. D10 values of Aeromonas cocktail in mixed sprouts, chicken and fish samples were found to be 0.081±0.001, 0.089±0.003 and 0.091±0.003 kGy, respectively. Radiation treatment with a 1.5 kGy dose resulted in complete elimination of 105 CFU/g of Aeromonas spp. from mixed sprouts, chicken and fish samples. No recovery of Aeromonas was observed in the 1.5 kGy treated samples stored at 4 °C up to 12 (mixed sprouts) and 7 days (chicken and fish samples), even after enrichment and selective plating. This study demonstrates that a 1.5 kGy dose of irradiation treatment could result in complete elimination of 105 CFU/g of Aeromonas spp. from mixed sprouts, chicken and fish samples.

Evaluation of different conditions and culture media for the recovery of Aeromonas spp. from water and shellfish samples.

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Latif-Eugenín, F; Beaz-Hidalgo, R; Figueras, M J

2016-09-01

To perform a comparative study for determining the optimum culture method (direct plating or enrichment) and medium (ampicillin dextrin agar (ADA), starch ampicillin agar (SAA), bile salts irgasan brilliant green modified (BIBG-m)) for recovering Aeromonas species from water and shellfish samples. By direct culture, Aeromonas was detected in 65% (13/20) of the water samples and in 54·5% (6/11) of the shellfish samples. However, when a pre-enrichment step was included, the number of positive water samples increased to 75% (15/20) and the ones of shellfish to 90·1% (10/11). The enriched culture significantly favoured (P culture medium for detecting Aeromonas from water was ADA. However, no differences were observed in the case of shellfish samples (P > 0·05). Isolation of Aeromonas media from water was favoured (P culture method and medium used influenced the recovery of some Aeromonas species from water and shellfish samples. This fact should be considered in future prevalence studies to avoid overestimating the above mentioned Aeromonas species. © 2016 The Society for Applied Microbiology.

Effectiveness of radiation processing in elimination of Aeromonas from food

International Nuclear Information System (INIS)

Nagar, Vandan; Bandekar, Jayant R.

2011-01-01

Genus Aeromonas has emerged as an important human pathogen because it causes a variety of diseases including gastroenteritis and extra-intestinal infections. Contaminated water, sprouts, vegetables, seafood and food of animal origin have been considered to be the important sources of Aeromonas infection. In the present study, radiation sensitivity of indigenous strains of Aeromonas spp. from different food samples was evaluated. The decimal reduction dose (D 10 ) values of different Aeromonas isolates in saline at 0-4 o C were in the range of 0.031-0.046 kGy. The mixed sprouts, chicken and fish samples were inoculated with a cocktail of five most resistant isolates (A. salmonicida Y567, A. caviae A85, A. jandaei A514A, A. hydrophila CECT 839 T and A. veronii Y47) and exposed to γ radiation to study the effectiveness of radiation treatment in elimination of Aeromonas. D 10 values of Aeromonas cocktail in mixed sprouts, chicken and fish samples were found to be 0.081±0.001, 0.089±0.003 and 0.091±0.003 kGy, respectively. Radiation treatment with a 1.5 kGy dose resulted in complete elimination of 10 5 CFU/g of Aeromonas spp. from mixed sprouts, chicken and fish samples. No recovery of Aeromonas was observed in the 1.5 kGy treated samples stored at 4 o C up to 12 (mixed sprouts) and 7 days (chicken and fish samples), even after enrichment and selective plating. This study demonstrates that a 1.5 kGy dose of irradiation treatment could result in complete elimination of 10 5 CFU/g of Aeromonas spp. from mixed sprouts, chicken and fish samples.

Effectiveness of radiation processing in elimination of Aeromonas from food

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Nagar, Vandan [Food Technology Division, Bhabha Atomic Research Centre, Mumbai 400085 (India); Bandekar, Jayant R., E-mail: jrb@barc.gov.i [Food Technology Division, Bhabha Atomic Research Centre, Mumbai 400085 (India)

2011-08-15

Genus Aeromonas has emerged as an important human pathogen because it causes a variety of diseases including gastroenteritis and extra-intestinal infections. Contaminated water, sprouts, vegetables, seafood and food of animal origin have been considered to be the important sources of Aeromonas infection. In the present study, radiation sensitivity of indigenous strains of Aeromonas spp. from different food samples was evaluated. The decimal reduction dose (D{sub 10}) values of different Aeromonas isolates in saline at 0-4 {sup o}C were in the range of 0.031-0.046 kGy. The mixed sprouts, chicken and fish samples were inoculated with a cocktail of five most resistant isolates (A. salmonicida Y567, A. caviae A85, A. jandaei A514A, A. hydrophila CECT 839{sup T} and A. veronii Y47) and exposed to {gamma} radiation to study the effectiveness of radiation treatment in elimination of Aeromonas. D{sub 10} values of Aeromonas cocktail in mixed sprouts, chicken and fish samples were found to be 0.081{+-}0.001, 0.089{+-}0.003 and 0.091{+-}0.003 kGy, respectively. Radiation treatment with a 1.5 kGy dose resulted in complete elimination of 10{sup 5} CFU/g of Aeromonas spp. from mixed sprouts, chicken and fish samples. No recovery of Aeromonas was observed in the 1.5 kGy treated samples stored at 4 {sup o}C up to 12 (mixed sprouts) and 7 days (chicken and fish samples), even after enrichment and selective plating. This study demonstrates that a 1.5 kGy dose of irradiation treatment could result in complete elimination of 10{sup 5} CFU/g of Aeromonas spp. from mixed sprouts, chicken and fish samples.

Biochemical characteristics, serogroups, and virulence factors of aeromonas species isolated from cases of diarrhoea and domestic water samples in Chennai

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Alavandi S

2003-01-01

Full Text Available PURPOSE: The objective of the present study was to delineate the differences between the clinical and environmental Aeromonas species with respect to their biochemical characteristics, serogrouping and virulence factors, in order to find a phenotypic marker of enteropathogenicity. METHODS: A total of 55 Aeromonas spp. inclusive of 19 isolates from cases of diarrhoea, and 36 from water samples comprising, 10 isolates of A. hydrophila, 21 isolates each of A. sobria, and A. caviae, two isolates of A. jandaei and one isolate of A. veronii were subjected to analysis of their biochemical characteristics, serogrouping, and virulence factors. RESULTS: Among the differences recorded in the biochemical characteristics in the three major species, the most striking characteristic was fermentation of lactose, which was observed in all the 11 A. caviae isolates recovered from water samples. None of the 10 clinical isolates of A. caviae tested fermented lactose. The clinical Aeromonas isolates belonged to seven typable serogroups, O:13, O:14, O:16, O:21, O:27, O:32 and O:35. The environmental isolates belonged to eight different serogroups, such as, O:3, O:11, O:14, O:16, O:18, O:28, O:64 and O:78 and were predominated by serotypes O:18 and O:64. Among the virulence factors tested, 89% of the environmental isolates produced b haemolysin, while only 62.3% of clinical isolates were able to do so. There was no significant difference between the clinical and environmental aeromonads with respect to their enterotoxigenicity in suckling mice in vivo, cytotoxicity in vitro in Vero cell monolayers, and ability to produce siderophores. CONCLUSION: Efforts to delineate the differences between the clinical and environmental Aeromonas spp. did not reveal significant difference between them. However, difference was observed with respect to their ability to produce b haemolysin, wherein, higher percentage of environmental isolates was haemolytic. The results also suggest

Chlorinated and ultraviolet radiation -treated reclaimed irrigation water is the source of Aeromonas found in vegetables used for human consumption

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Latif-Eugenín, Fadua; Beaz-Hidalgo, Roxana; Silvera-Simón, Carolina [Unidad de Microbiología, Facultad de Medicina y Ciencias de la Salud, IISPV, Universidad Rovira i Virgili, Reus (Spain); Fernandez-Cassi, Xavi [Departamento de Microbiología, Facultad de Biología, Universidad de Barcelona, Barcelona (Spain); Figueras, María J., E-mail: mariajose.figueras@urv.cat [Unidad de Microbiología, Facultad de Medicina y Ciencias de la Salud, IISPV, Universidad Rovira i Virgili, Reus (Spain)

2017-04-15

Wastewater is increasingly being recognized as a key water resource, and reclaimed water (or treated wastewater) is used for irrigating vegetables destined for human consumption. The aim of the present study was to determine the diversity and prevalence of Aeromonas spp. both in reclaimed water used for irrigation and in the three types of vegetables irrigated with that water. Seven of the 11 (63.6%) samples of reclaimed water and all samples of vegetables were positive for the presence of Aeromonas. A total of 216 Aeromonas isolates were genotyped and corresponded to 132 different strains that after identification by sequencing the rpoD gene belonged to 10 different species. The prevalence of the species varied depending on the type of sample. In the secondary treated reclaimed water A. caviae and A. media dominated (91.4%) while A. salmonicida, A. media, A. allosaccharophila and A. popoffii represented 74.0% of the strains in the irrigation water. In vegetables, A. caviae (75.0%) was the most common species, among which a strain isolated from lettuce had the same genotype (ERIC pattern) as a strain recovered from the irrigation water. Furthermore, the same genotype of the species A. sanarellii was recovered from parsley and tomatoes demonstrating that irrigation water was the source of contamination and confirming the risk for public health. - Highlights: • Reclaimed water (= treated wastewater) is used for the irrigation of vegetables. • Aeromonas was found in reclaimed water and irrigated vegetables with this water. • The prevalence of Aeromonas spp. varied between irrigation water and vegetables. • Epidemiological relationships were found between irrigation water and vegetables. • The water was the source of contamination which means a risk for the public health.

Chlorinated and ultraviolet radiation -treated reclaimed irrigation water is the source of Aeromonas found in vegetables used for human consumption

International Nuclear Information System (INIS)

Latif-Eugenín, Fadua; Beaz-Hidalgo, Roxana; Silvera-Simón, Carolina; Fernandez-Cassi, Xavi; Figueras, María J.

2017-01-01

Wastewater is increasingly being recognized as a key water resource, and reclaimed water (or treated wastewater) is used for irrigating vegetables destined for human consumption. The aim of the present study was to determine the diversity and prevalence of Aeromonas spp. both in reclaimed water used for irrigation and in the three types of vegetables irrigated with that water. Seven of the 11 (63.6%) samples of reclaimed water and all samples of vegetables were positive for the presence of Aeromonas. A total of 216 Aeromonas isolates were genotyped and corresponded to 132 different strains that after identification by sequencing the rpoD gene belonged to 10 different species. The prevalence of the species varied depending on the type of sample. In the secondary treated reclaimed water A. caviae and A. media dominated (91.4%) while A. salmonicida, A. media, A. allosaccharophila and A. popoffii represented 74.0% of the strains in the irrigation water. In vegetables, A. caviae (75.0%) was the most common species, among which a strain isolated from lettuce had the same genotype (ERIC pattern) as a strain recovered from the irrigation water. Furthermore, the same genotype of the species A. sanarellii was recovered from parsley and tomatoes demonstrating that irrigation water was the source of contamination and confirming the risk for public health. - Highlights: • Reclaimed water (= treated wastewater) is used for the irrigation of vegetables. • Aeromonas was found in reclaimed water and irrigated vegetables with this water. • The prevalence of Aeromonas spp. varied between irrigation water and vegetables. • Epidemiological relationships were found between irrigation water and vegetables. • The water was the source of contamination which means a risk for the public health.

Biological control of Aeromonas salmonicida subsp. salmonicida infection in rainbow trout (Oncorhynchus mykiss) using Aeromonas phage PAS-1.

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Kim, J H; Choresca, C H; Shin, S P; Han, J E; Jun, J W; Park, S C

2015-02-01

The potential control efficacy of Aeromonas phage PAS-1 was evaluated against Aeromonas salmonicida subsp. salmonicida infection in rainbow trout (Oncorhynchus mykiss) model in this study. The phage was co-cultured with the virulent A. salmonicida subsp. salmonicida strain AS05 that possesses the type III secretion system (TTSS) ascV gene, and efficient bacteriolytic activity was observed against the bacteria. The administration of PAS-1 in rainbow trout demonstrated that the phage was cleared from the fish within 200 h post-administration, and a temporal neutralizing activity against the phage was detected in the sera of phage-administrated fish. The administration of PAS-1 (multiplicity of infection: 10 000) in A. salmonicida subsp. salmonicida infected rainbow trout model showed notable protective effects, with increased survival rates and mean times to death. These results demonstrated that Aeromonas phage PAS-1 could be considered as an alternative biological control agent against A. salmonicida subsp. salmonicida infections in rainbow trout culture. © 2013 Blackwell Verlag GmbH.

Occurrence of potential pathogenic Aeromonas species in tropical seafood, aquafarms and mangroves off Cochin coast in South India

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Alphonsa Vijaya Joseph

2013-12-01

Full Text Available Background: The genus Aeromonas include gram-negative, motile, facultative anaerobic, rod shaped and oxidase positive bacteria comprising several species, associated with the aquatic environment. Aeromonas species have been implicated in human pathogenesis and are linked with gastroenteritis, muscle infections, septicemia, and skin diseases. In fish they are renowned as enteric pathogens causing haemorrhagic septicemia, fin rot, soft tissue rot and furunculosis resulting in major die-offs and fish kills. Aim: This study reports the occurrence of potential pathogenic Aeromonas sp. in tropical seafood (Squids, Prawns and Mussels, aquafarms and mangroves of Cochin, Kerala, South India. Materials and Methods :Tropical seafood (Squid, Prawn and Mussel, sediment and water samples from aquafarms and associated mangroves were screened for Aeromonas contamination. The isolates were identified by 16S rDNA sequence analysis and subjected to morphological and biochemical characterization. Haemolytic assay was used for determining pathogenicity of the organisms. Antibiotic susceptibility against 12 antibiotics were performed and the MAR index was calculated. Results: A total of 134 isolates were recovered from the samples of which 15 were identified as Aeromonas species by 16S rDNA sequence analysis and were assigned to 5 species namely, A. hydrophila, A. enteropelogenes, A. caviae, A. punctataand A. aquarorium. Morphological, biochemical and phylogenetic analyses revealed relatedness and variability among the strains. All the isolates were haemolytic on blood agar indicating their pathogenicity. The isolates exhibited varying degrees of resistance to vancomycin (86.66%, ampicillin (46.66%, nalidixic acid (20%, tetracycline (6.66%, co-trimaxozole (6.66% and rifampicin (6.66% and were susceptible to antibiotics like gentamycin, streptomycin, trimethoprim, azithromycin, cefixime and chloramphenicol. 20% of Aeromonas sp. showed MAR index > 0.2 indicative of the

Species Distribution and Prevalence of Putative Virulence Factors in Mesophilic Aeromonas spp. Isolated from Fresh Retail Sushi

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Sunniva Hoel

2017-05-01

Full Text Available Aeromonas spp. are ubiquitous bacteria that have received increasing attention as human pathogens because of their widespread occurrence in food, especially seafood and vegetables. The aim of this work was to assess the species identity and phylogenetic relationship of 118 Aeromonas strains isolated from fresh retail sushi from three producers, and to characterize the isolates with respect to genetic and phenotypic virulence factors. We also evaluate the potential hazard associated with their presence in ready-to-eat seafood not subjected to heat treatment. Mesophilic Aeromonas salmonicida was most prevalent (74%, followed by A. bestiarum (9%, A. dhakensis (5%, A. caviae (5%, A. media (4%, A. hydrophila (2%, and A. piscicola (1%. All isolates were considered potentially pathogenic due to the high prevalence of genes encoding hemolysin (hlyA (99%, aerolysin (aerA (98%, cytotoxic enterotoxin (act (86%, heat-labile cytotonic enterotoxin (alt (99%, and heat-stable cytotonic enterotoxin (ast (31%. The shiga-like toxins 1 and 2 (stx-1 and stx-2 were not detected. Moreover, there was heterogeneity in toxin gene distribution among the isolates, and the combination of act/alt/hlyA/aerA was most commonly detected (63%. β-hemolysis was species-dependent and observed in 91% of the isolates. All A. media and A. caviae strains were non-hemolytic. For isolates belonging to this group, lack of hemolysis was possibly related to the absence of the act gene. Swimming motility, linked to adhesion and host invasion, occurred in 65% of the isolates. Partial sequencing of the gyrB gene demonstrated its suitability as a genetic marker for Aeromonas species identification and for assessment of the phylogenetic relationship between the isolates. The gyrB sequence divergence within a given species ranged from 1.3 to 2.9%. A. bestiarum, A. salmonicida, and A. piscicola were the most closely related species; their sequences differed by 2.7–3.4%. The average gyrB sequence

Prevalência de Aeromonas ssp em fezes diarréicas de crianças menores de 5 anos de idade na cidade de Goiânia, Goiás, no biênio 1995 - 1996

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Idalina Thiomi Inumaru Nojimoto

1997-10-01

Full Text Available Foram analisadas 163 amostras de fezes de crianças com idade abaixo de 5 anos no período de 1995 a 1996, sendo 91 de fezes diarréicas e 72 de fezes não diarréicas. O material foi coletado em meio para transporte e submetido ao processo de enriquecimento a 4oC por 7 dias. Para o isolamento primário foi utilizado ágar amido ampicilina e incubado a 35oC por 18 a 24 horas. Foram isoladas 20 (21,9% das seguintes espécies: Aeromonas A. caviae (7,7%, A. salmonicida salmonicida (6,6%, A. sobria (4,3%, A. hydrophila (2,2% e Salmonicida achromogenes (1,1%. Nenhuma Aeromonas spp foi isolada dos 72 pacientes-controles. A susceptibilidade das amostras de Aeromonas spp aos antimicrobianos foi maior com a ciprofloxacina, diminuindo gradativamente com cloranfenicol, gentamicina, ampicilina e eritromicina.From 1995 through 1996, 163 fecal specimens of children aged under 5 years were analysed, 91 being from diarrhea feces and 72 without diarrhea. The material was collected in transport medium and submitted to the enrichment procedure at 4oC for 7 days. For the primary isolation starch ampicillin agar was used and incubated at 35oC for 18 to 24 hours. Twenty (20.9% from the following specimens were isolated: Aeromonas A. caviae (7.7%, A. salmonicida salmonicida (6.6%, A. sobria (4.3%, A. hydrophila (2.2% e Salmonicida achromogenes (1.1%. No Aeromonas spp was isolated from the 72 control subjects. The Aeromonas spp susceptibility to antimicrobian was greater with ciprofloxacin, being this susceptibility gradually diminished with chloranphenicol, gentamicin, ampicillin and erythromycin.

Aeromonas spp.: an emerging pathogen?

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Andrea Bartolini

2015-12-01

Full Text Available The aim of this study is to identify and monitor the presence of Aeromonas spp. strains in stool cultures. We analyzed 5564 stool cultures from September 2012 to August 2013. Sixty-three patients were positive for Aeromonas spp. The most frequent symptoms were: diarrhea (46.0% and abdominal pain (12.7%. Pediatric subjects were 28. Samples’ microscopic examination showed leukocytes in 38.1% of cases. It is still controversial whether Aeromonas are responsible for human gastroenteritis, but their presence in faecies of symptomatic patients supports their etiologic role. We propose search for toxins by polymerase chain reaction to identify strains that require an antibiotic therapy.

Genospecies and virulence factors of Aeromonas species in different sources in a North African country

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Khalifa Sifaw Ghenghesh

2014-09-01

Full Text Available Introduction: Aeromonads of medical importance have been reported from numerous clinical, food, and water sources, but identification of genospecies and virulence factors of Aeromonas species from countries in North Africa and the Middle East are few. Methods: In total 99 Aeromonas species isolates from different sources (diarrheal children [n=23], non-diarrheal children [n=16], untreated drinking water from wells [n=32], and chicken carcasses [n=28] in Tripoli, Libya, were included in the present investigation. Genus identification was confirmed by biochemical analysis, and genospecies were determined using a combination of 16S rDNA variable region and gyrB sequence analysis. Polymerase chain reaction (PCR was used to detect genes encoding toxins from 52 of the isolates. Results: We identified 44 isolates (44% as A. hydrophila (3 [3.0%] subspecies anaerogenes, 23 [23%] subspecies dhakensis, and 18 [18%] subspecies ranae; 27 isolates (27% as A. veronii; 23 isolates (23% as A. caviae; and 5 isolates (5.0% as other genospecies. The genes encoding aerolysin (aer, cytolytic enterotoxin (act, and A. hydrophila isolate SSU enterotoxin (ast were detected in 45 (87%, 4 (7.7%, and 9 (17% of the 52 isolates tested, respectively. The gene encoding an extracellular lipase (alt was not detected. Conclusion: The majority of aeromonads from Libya fall within three genospecies (i.e. A. hydrophila, A. veronii, and A. caviae, and genes coding for toxin production are common among them.

Clinical manifestations of bacteremia caused by Aeromonas species in southern Taiwan.

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Hung-Jen Tang

Full Text Available This study is conducted to investigate the clinical characteristics of patients with bacteremia caused by Aeromonas species.Patients with bacteremia caused by Aeromonas species during the period 2009 to 2013 were identified from a computerized database of a regional hospital in southern Taiwan. The medical records of these patients were retrospectively reviewed.A total of 91 patients with bacteremia due to Aeromonas species were identified. In addition to 16 (17.6% primary bacteremia, the most common source of secondary infection is peritonitis (n = 27, 29.7%, followed by biliary tract infection (n = 18, 19.8%, and SSTI (n = 12, 13.2%, pneumonia (n = 9, 9.9%, catheter-related bloodstream infection (n =  5, 5.5%, and genitourinary tract infection (n = 4, 4.4%. A. hydrophila (n = 35, 38.5% was the most common pathogen, followed by A. veronii biovar sobria (n = 31, 34.1%, A. caviae (n = 14, 15.4%, and A. veronii biovar veronii (n = 9, 9.9%. Forty-three (47.3% patients were classified as healthcare-associated infections (HCAI causes by Aeromonas species, and patients with HCAI were more likely to have cancer, and receive immunosuppressant than patients with community-acquired bacteremia. The overall outcomes, including rate of ICU admission, acute respiratory failure, and mortality were 33.3%, 28.6%, and 23.1%, respectively. Multivariate analysis showed that the in-hospital day mortality was significantly associated only with underlying cancer (P <.001, and initial shock (P <.001.Aeromonas species should be considered one of the causative pathogens of healthcare-associated bacteremia, especially in immunocompromised patients. In addition, it can be associated with high fatality. Cancer and initial shock were the poor prognostic factors.

Prevalence of fur mites (Chirodiscoides caviae) in pet guinea pigs (Cavia porcellus) in southern Italy.

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d'Ovidio, Dario; Santoro, Domenico

2014-04-01

Chirodiscoides caviae is the most common fur mite affecting guinea pigs; infestation is generally asymptomatic. No studies have been published on the prevalence of such mites in guinea pigs in southern Italy. We sought to evaluate the prevalence and the clinical signs of C. caviae infestation in guinea pigs in southern Italy. Clinical records of guinea pigs evaluated from August 2012 to July 2013 were retrospectively searched. In this retrospective matched case-control study, records of guinea pigs with evidence of C. caviae infestation were selected. The prevalence of C. caviae infestation was evaluated and exposure variables were assessed among guinea pigs with and without infestation using stepwise conditional logistic regression. Guinea pigs seen during the same time period, but without a diagnosis of C. caviae, were included as control animals. The prevalence of C. caviae was 32% (42 of 131); 66.6% of affected guinea pigs (28 of 42) originated from pet shops, whereas 28% (14 of 42) were privately owned. Thirty-one guinea pigs (73.8%) were asymptomatic, whereas 11 (26.1%) showed clinical signs (pruritus, alopecia, erythema and scaling). The most frequently affected area was the lumbosacral region (38 of 42). Guinea pigs in pet shops were more likely to be affected by C. caviae than owned guinea pigs (odds ratio, 5.12; 95% confidence interval, 2.32-11.29; P guinea pigs in southern Italy. Chirodiscoides mites should be sought in guinea pigs, particularly in animals coming from pet shops. © 2014 ESVD and ACVD.

A molecular study on the prevalence and virulence potential of Aeromonas spp. recovered from patients suffering from diarrhea in Israel.

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Yigal Senderovich

Full Text Available BACKGROUND: Species of the genus Aeromonas are native inhabitants of aquatic environments and have recently been considered emerging human pathogens. Although the gastrointestinal tract is by far the most common anatomic site from which aeromonads are recovered, their role as etiologic agents of bacterial diarrhea is still disputed. Aeromonas-associated diarrhea is a phenomenon occurring worldwide; however, the exact prevalence of Aeromonas infections on a global scale is unknown. METHODOLOGY/PRINCIPAL FINDINGS: The prevalence and virulence potential of Aeromonas in patients suffering from diarrhea in Israel was studied using molecular methods. 1,033 diarrheal stools were sampled between April and September 2010 and Aeromonas species were identified in 17 (∼2% patients by sequencing the rpoD gene. Aeromonas species identity and abundance was: A. caviae (65%, A. veronii (29% and Aeromonas taiwanensis (6%. This is the first clinical record of A. taiwanensis as a diarrheal causative since its recent discovery from a wound infection in a patient in Taiwan. Most of the patients (77% from which Aeromonas species were isolated were negative for any other pathogens. The patients ranged from 1 to 92 years in age. Aeromonas isolates were found to possess different virulence-associated genes: ahpB (88%, pla/lip/lipH3/apl-1 (71%, act/hlyA/aerA (35%, alt (18%, ast (6%, fla (65%, lafA (41%, TTSS ascV (12%, TTSS ascF-ascG (12%, TTSS-dependent ADP-ribosylating toxins aexU (41% and aexT (6% in various combinations. Most of the identified strains were resistant to beta-lactam antibiotics but susceptible to third-generation cephalosporin antibiotics. CONCLUSIONS: Aeromonas may be a causative agent of diarrhea in patients in Israel and therefore should be included in routine bacteriological screenings.

Phenotypic and Genetic Diversity of Aeromonas Species Isolated from Fresh Water Lakes in Malaysia.

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Wei Ching Khor

Full Text Available Gram-negative bacilli of the genus Aeromonas are primarily inhabitants of the aquatic environment. Humans acquire this organism from a wide range of food and water sources as well as during aquatic recreational activities. In the present study, the diversity and distribution of Aeromonas species from freshwater lakes in Malaysia was investigated using glycerophospholipid-cholesterol acyltransferase (GCAT and RNA polymerase sigma-factor (rpoD genes for speciation. A total of 122 possible Aeromonas strains were isolated and confirmed to genus level using the API20E system. The clonality of the isolates was investigated using ERIC-PCR and 20 duplicate isolates were excluded from the study. The specific GCAT-PCR identified all isolates as belonging to the genus Aeromonas, in agreement with the biochemical identification. A phylogenetic tree was constructed using the rpoD gene sequence and all 102 isolates were identified as: A. veronii 43%, A. jandaei 37%, A. hydrophila 6%, A. caviae 4%, A. salmonicida 2%, A. media 2%, A. allosaccharophila 1%, A. dhakensis 1% and Aeromonas spp. 4%. Twelve virulence genes were present in the following proportions--exu 96%, ser 93%, aer 87%, fla 83%, enolase 70%, ela 62%, act 54%, aexT 33%, lip 16%, dam 16%, alt 8% and ast 4%, and at least 2 of these genes were present in all 102 strains. The ascV, aexU and hlyA genes were not detected among the isolates. A. hydrophila was the main species containing virulence genes alt and ast either present alone or in combination. It is possible that different mechanisms may be used by each genospecies to demonstrate virulence. In summary, with the use of GCAT and rpoD genes, unambiguous identification of Aeromonas species is possible and provides valuable data on the phylogenetic diversity of the organism.

Potential enterotoxicity and antimicrobial resistance pattern of Aeromonas species isolated from pet turtles and their environment.

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Wimalasena, S H M P; Shin, Gee-Wook; Hossain, Sabrina; Heo, Gang-Joon

2017-05-23

To investigate the potential enterotoxicity and antimicrobial resistance of aeromonads from pet turtles as a risk for human infection, one hundred and two Aeromonas spp. were isolated from the feces, skin and rearing environments of pet turtles and identified by biochemical and gyrB sequence analyses. Aeromonas enteropelogenes was the predominant species among the isolates (52.9%) followed by A. hydrophila (32.4%), A. dharkensis (5.9%), A. veronii (4.9%) and A. caviae (3.9%). Their potential enterotoxicities were evaluated by PCR assays for detecting genes encoding cytotoxic enterotoxin (act) and two cytotonic enterotoxins (alt and ast). 75.8% of A. hydrophila isolates exhibited the act + /alt + /ast + genotype, whereas 94.4% of A. enteropelogenes isolates were determined to be act - /alt - /ast - . In an antimicrobial susceptibility test, most isolates were susceptible to all tested antibiotics except amoxicillin, ampicillin, cephalothin, chloramphenicol and tetracycline. Non-susceptible isolates to penicillins (ampicillin and amoxicillin) and fluoroquinolones (ciprofloxacin and norfloxacin) were frequently observed among the A. enteropelogenes isolates. Few isolates were resistant to imipenem, amikacin, ceftriaxone and cefotaxime. Collectively, these results suggest that pet turtles may pose a public health risk of infection by enterotoxigenic and antimicrobial resistant Aeromonas strains.

İnek Sütlerinin Hareketli Aeromonas Türleri Yönünden İncelenmesi

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Mustafa Alişarlı

2015-02-01

Full Text Available Bu çalışmada, çiğ sütlerde hareketli Aermonas türlerinin bulunma sıklığını ortaya koymak ve ilk sağılan sütün kontaminasyondaki rolünü belirlemek amaçlanmaştır. Bu amaçla, 100 adet sağmal inekten direkt el sağımı ile alınan süt örnekleri incelenmiştir. İncelenen Örneklerin 8 tanesinde (%8 Aeromonas spp.izole edilmiş olup bunlardan 3'ü (%37,5 hareketli Aeromonas olarak belirlenmiştir. Hareketli Aeromonas'ların 2'si (%66,6 A. hydrophila ve1'i (%33,3 A. caviae olarak identifiye edilmiştir. Bu araştırmanın verileri, süt işletmelerine tanker ve güğümlerle getirilen çiğ sütlerden sağlanan örneklerdeki araştırma bulgularıyla kıyaslandığında daha düşük bulunmuştur. Bu da, eğer sağım sonrası hijyen kurallarına gerekli özen gösterilmezse çiğ sütlerin Aeromonas'lar ile kontaminasyon riskinin yüksek olacağını göstermektedir.

Aeromonas spp. e Plesiomonas shigelloides isoladas a partir de mexilhões (Perna perna in natura e pré-cozidos no Rio de Janeiro, RJ Aeromonas spp. and Plesiomonas shigelloides isolated from in natura and precooked mussels (Perna perna in Rio de Janeiro, RJ, Brazil

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Christiane Soares Pereira

2004-12-01

Full Text Available O ecossistema aquático é o habitat de mexilhões (Perna perna, animais filtradores que refletem a qualidade ambiental através de análise microbiológica de sua carne. No presente trabalho avaliou-se a presença de patógenos emergentes (Aeromonas hydrophila e Plesiomonas shigelloides, em mexilhões in natura e pré-cozidos coletados por pescadores da Estação Experimental de Cultivo de Mexilhões situada em Jurujuba, Niterói, Rio de Janeiro. Foram analisadas 86 amostras de mexilhões (43 in natura e 43 pré-cozidos as quais foram submetidas a enriquecimento em Água Peptonada Alcalina (APA acrescida de 1 e 3% de Cloreto de Sódio (NaCl e em solução Salina de Butterfield, incubadas a 37ºC por 24 horas. Em seguida, foram semeadas em Ágar Seletivo para Pseudomonas-Aeromonas (GSP, Ágar Tiossulfato Citrato Bile Sacarose (TCBS e Ágar Inositol Bile Verde Brilhante (IBB. A análise geral dos resultados permitiu a identificação de Areomonas spp e Plesiomonas shigelloides em 86% das amostras de mexilhões in natura e pré-cozidas avaliadas. A posterior caracterização bioquímica permitiu a identificação das espécies Aeromonas media (37,10%, A. hydrophila (15,50%, A. caviae (14,80%, A. veronii biogrupo veronii (11,60%, Aeromonas sp. (7,36%, A. sobria (4,20%, A. trota (4,20%, A. schubertii (1,31%, A. jandaei (1,31%, A. veronii biogrupo sobria (0,52% e Plesiomonas shigelloides (2,10%. A relevância epidemiológica desses microrganismos em casos de gastrenterite humana, após consumo de mexilhões crus ou parcialmente cozidos, revela a importância de alertar as autoridades de Saúde Pública no Brasil, sobre a presença desses patógenos na cadeia alimentar e seus riscos para a saúde humana.The aquatic ecosystem is the habitat of mussels, filtrating animals that reflect the ambient quality through microbiological analysis. In the present investigation, we evaluated the presence of emergent pathogens (Aeromonas hydrophila and Plesiomonas

Cavia porcellus

African Journals Online (AJOL)

SARAH

30 sept. 2017 ... quality during early development on body weight and reproductive maturation of guinea pigs. (Cavia porcellus). Gen. Comp. Endocrin. 161,. 384-389 ;. Chauca de Zaldivar 1999 Produccion de cuyes en crianzas familiares en Peru. In : V Congreso. Latino americano de Cuyicultura, Venezuela,. Memorias…

Chlamydia caviae infection alters abundance but not composition of the guinea pig vaginal microbiota.

Science.gov (United States)

Neuendorf, Elizabeth; Gajer, Pawel; Bowlin, Anne K; Marques, Patricia X; Ma, Bing; Yang, Hongqiu; Fu, Li; Humphrys, Michael S; Forney, Larry J; Myers, Garry S A; Bavoil, Patrik M; Rank, Roger G; Ravel, Jacques

2015-06-01

In humans, the vaginal microbiota is thought to be the first line of defense again pathogens including Chlamydia trachomatis. The guinea pig has been extensively used as a model to study chlamydial infection because it shares anatomical and physiological similarities with humans, such as a squamous vaginal epithelium as well as some of the long-term outcomes caused by chlamydial infection. In this study, we aimed to evaluate the guinea pig-C. caviae model of genital infection as a surrogate for studying the role of the vaginal microbiota in the early steps of C. trachomatis infection in humans. We used culture-independent molecular methods to characterize the relative and absolute abundance of bacterial phylotypes in the guinea pig vaginal microbiota in animals non-infected, mock-infected or infected by C. caviae. We showed that the guinea pig and human vaginal microbiotas are of different bacterial composition and abundance. Chlamydia caviae infection had a profound effect on the absolute abundance of bacterial phylotypes but not on the composition of the guinea pig vaginal microbiota. Our findings compromise the validity of the guinea pig-C. caviae model to study the role of the vaginal microbiota during the early steps of sexually transmitted infection. © FEMS 2015.

Assessment of toxicity of a glyphosate-based formulation using bacterial systems in lake water.

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Amorós, I; Alonso, J L; Romaguera, S; Carrasco, J M

2007-05-01

A new Aeromonas bioassay is described to assess the potential harmful effects of the glyphosate-based herbicide, Roundup, in the Albufera lake, a protected area near Valencia. Viability markers as membrane integrity, culturability and beta-galactosidase production of Aeromonas caviae were studied to determine the influence of the herbicide in the bacterial cells. Data from the multifactor analysis of variance test showed no significant differences (P>0.05) between A. caviae counts of viability markers at the studied concentrations (0, 50 and 100 mg l-1 of glyphosate). The effects of Roundup on microbial biota present in the lake were assessed by measuring the number of indigenous mesophilic Aeromonas in presence of different amounts of the herbicide at 0, 50 and 100 mg l-1 of glyphosate. In samples containing 50 and 100 mg l-1 of glyphosate a significant (PAlbufera lake water to Microtox luminescent bacterium (Vibrio fischeri) also was determined. The EC50 values obtained were 36.4 mg l-1 and 64.0 mgl-1 of glyphosate respectively. The acidity (pH 4.5) of the herbicide formulation was the responsible of the observed toxicity.

Aeromonas Bacteraemia Causing Gas Gangrene in a Diabetic ...

African Journals Online (AJOL)

Early blood culture, culture of the aspirates from blebs, and subsequent culture of intraoperative specimens uniformly grew Aeromonas hydrophila. Urgent fasciotomy was carried out same day, and the patient was admitted into ICU for intensive monitoring and therapies. Second look procedures 48hours later showed turgid ...

The involvement of tetA and tetE tetracycline resistance genes in plasmid and chromosomal resistance of Aeromonas in Brazilian strains

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Ilana Teruszkin Balassiano

2007-11-01

Full Text Available This study analyzed the involvement of tetA and tetE genes in the tetracycline resistance of 16 strains of genus Aeromonas, isolated from clinical and food sources. Polymerase chain reactions revealed that 37.5% of the samples were positive for tetA, and also 37.5% were tetE positive. One isolate was positive for both genes. Only the isolate A. caviae 5.2 had its resistance associated to the presence of a plasmid, pSS2. The molecular characterization of pSS2 involved the construction of its restriction map and the determination of its size. The digestion of pSS2 with HindIII originated two fragments (A and B that were cloned separately into the pUC18 vector. The tetA gene was shown to be located on the HindIII-A fragment by PCR. After transforming a tetracycline-sensitive strain with pSS2, the transformants expressed the resistance phenotype and harbored a plasmid whose size was identical to that of pSS2. The results confirmed the association between pSS2 and the tetracycline resistance phenotype, and suggest a feasible dissemination of tetA and tetE among strains of Aeromonas. This study suggests the spreading tetA and tetE genes in Aeromonas in Brazil and describes a resistance plasmid that probably contributes to the dissemination of the resistance.

Functional genomic characterization of virulence factors from necrotizing fasciitis-causing strains of Aeromonas hydrophila.

Science.gov (United States)

Grim, Christopher J; Kozlova, Elena V; Ponnusamy, Duraisamy; Fitts, Eric C; Sha, Jian; Kirtley, Michelle L; van Lier, Christina J; Tiner, Bethany L; Erova, Tatiana E; Joseph, Sandeep J; Read, Timothy D; Shak, Joshua R; Joseph, Sam W; Singletary, Ed; Felland, Tracy; Baze, Wallace B; Horneman, Amy J; Chopra, Ashok K

2014-07-01

The genomes of 10 Aeromonas isolates identified and designated Aeromonas hydrophila WI, Riv3, and NF1 to NF4; A. dhakensis SSU; A. jandaei Riv2; and A. caviae NM22 and NM33 were sequenced and annotated. Isolates NF1 to NF4 were from a patient with necrotizing fasciitis (NF). Two environmental isolates (Riv2 and -3) were from the river water from which the NF patient acquired the infection. While isolates NF2 to NF4 were clonal, NF1 was genetically distinct. Outside the conserved core genomes of these 10 isolates, several unique genomic features were identified. The most virulent strains possessed one of the following four virulence factors or a combination of them: cytotoxic enterotoxin, exotoxin A, and type 3 and 6 secretion system effectors AexU and Hcp. In a septicemic-mouse model, SSU, NF1, and Riv2 were the most virulent, while NF2 was moderately virulent. These data correlated with high motility and biofilm formation by the former three isolates. Conversely, in a mouse model of intramuscular infection, NF2 was much more virulent than NF1. Isolates NF2, SSU, and Riv2 disseminated in high numbers from the muscular tissue to the visceral organs of mice, while NF1 reached the liver and spleen in relatively lower numbers on the basis of colony counting and tracking of bioluminescent strains in real time by in vivo imaging. Histopathologically, degeneration of myofibers with significant infiltration of polymorphonuclear cells due to the highly virulent strains was noted. Functional genomic analysis provided data that allowed us to correlate the highly infectious nature of Aeromonas pathotypes belonging to several different species with virulence signatures and their potential ability to cause NF. Copyright © 2014, American Society for Microbiology. All Rights Reserved.

The Influences of Bacillus subtilis on the Virulence of Aeromonas hydrophila and Expression of luxS Gene of Both Bacteria Under Co-cultivation.

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Ren, Yuwei; Li, Sisi; Wu, Zhixin; Zhou, Chengchong; Zhang, Ding; Chen, Xiaoxuan

2017-06-01

The aim of this study was to explore the influence of Bacillus subtilis CH9 on Aeromonas hydrophila SC2005. The transcription level of virulence genes of A. hydrophila SC2005 and its hemolysin activity as well as its cytotoxicity were analyzed when B. subtilis CH9 and A. hydrophila SC2005 were co-cultured. The results indicated that the transcription levels of four virulence genes of A. hydrophila, including aer, ahyB, hcp, and emp, decreased when A. hydrophila was cultured with B. subtilis CH9. Furthermore, the extracellular products of A. hydrophila showed attenuated hemolysin activity as well as cytotoxicity when A. hydrophila was cultured with B. subtilis CH9. Finally, the transcriptional levels of luxS genes of B. subtilis CH9 and A. hydrophila SC2005 were determined when these two species were co-cultured. RT-qPCR results suggested that the transcription level of A. hydrophila was down-regulated significantly. On the contrary, the transcription level of B. subtilis CH9 was up-regulated significantly. These results suggested that the probiotic role of B. subtilis CH9 is related to the inhibition of growth and virulence of A. hydrophila SC2005, and quorum sensing may be involved.

Identificação e resistência a antimicrobianos de espécies de Aeromonas móveis isoladas de peixes e ambientes aquáticos Identification and antimicrobial resistance of motile Aeromonas isolated from fish and aquatic environment

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Daniela Hirsch

2006-12-01

Full Text Available Com o objetivo de se verificar a diversidade de espécies de Aeromonas móveis e seu perfil de susceptibilidade a antimicrobianos em pisciculturas comerciais, foram selecionadas oito tilapiculturas localizadas na região do Alto Rio Grande, Minas Gerais. De cada propriedade foram coletadas três amostras de peixes em estádio de pré-abate (vivos e saudáveis, uma amostra de água do tanque e uma amostra da água de abastecimento do sistema. De cada peixe foram coletadas amostras de lavado superficial e do parênquima renal. Diluições seriadas adequadas de cada amostra foram plaqueadas em TSA-ampicilina (10 mg/l e as amostras de rim em Ágar Sangue de cavalo a 5%. A partir de colônias isoladas positivas para o teste da oxidase foram realizados testes para identificação do gênero (testes presuntivos e das espécies de Aeromonas (testes bioquímicos. O perfil de antibiograma foi obtido pelo teste de difusão de discos de antibióticos em Ágar Mueller Hinton. Foram obtidos 75 isolados diferenciados em nove espécies de Aeromonas: A. jandaei, A. hydrophila, A. trota, A. caviae, A. sobria, A. eucrenophila, A. veronii bt veronii, A. schubertii, A. media, além de amostras classificadas como Aeromonas atípicas. Do total isolado, oito amostras foram provenientes da superfície corpórea de peixes, 14 da água de abastecimento e 53 da água do tanque. Não houve isolamento a partir dos espécimes de parênquima renal. Em relação ao perfil de resistência, 93% dos isolados foram resistentes à eritromicina, 36% à tetraciclina, 13% ao ác. nalidíxico, 9% à gentamicina, 8% à nitrofurantoína, 8% à canamicina, 5% à norfloxacina, 4% ao cloranfenicol e 3% às sulfonamidas. Dentre os isolados analisados, 43% apresentaram índice de múltipla resistência a antimicrobianos (MAR igual ou superior a 22%, ou seja, resistência a dois ou mais drogas das nove testadas. Os dados apontam para um risco iminente, tanto pelo isolamento de amostras

Aeromonas veronii septicemia in an immunocompetent patient

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Paula McKenzie

2013-09-01

Full Text Available We present a 29-year-old healthy man who fell into an industrial auger, sustaining acrushed, open pelvic injury, multiple comminuted fractures of the right leg, and traumaticamputation of his left foot. Blood and wound cultures were positive for Aeromonasspp and vancomycin resistant Enterococcus. Treatment included cefepime, levofloxacin,daptomycin, and metronidazole. Aeromonas veronii is a Gram negative bacillus usuallyfound in fresh and brackish water in warm climates. It can cause severe skin andsoft tissue infections, typically after injured tissue is exposed to contaminated water.Aeromonas septicemia is uncommon and is usually associated with underlying diseases,such as malignancy, cirrhosis, diabetes, or immunosuppression. It rarely occurs in ahealthy host.

First isolation of Aeromonas salmonicida subspecies salmonicida from diseased sea bass, Dicentrarchus labrax (L., cultured in Spain

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Clara Fernández-Álvarez

2016-11-01

Full Text Available This work represents the first description of Aeromonas salmonicida subsp. salmonicida as causative agent of furunculosis in cultured sea bass, Dicentrarchus labrax (L.. Cumulative mortality in affected fish from two floating cages in the Mediterranean coast of Spain was 3.8%. Affected sea bass did not show the typical external signs of furunculosis in the first stages of the disease, however, when the disease progressed, open ulcers appeared on the skin and muscle. Internally, splenomegaly was the only pathological sign observed. Samples from diseased fish were subjected to standardized assays for pathogens screening. Negative results were obtained for parasites and fish viruses. A Gram-stain-negative rod-shaped bacterium was observed in smears from liver, kidney and spleen of all analysed fish. Pure bacterial cultures were recovered from liver, kidney and spleen of all diseased fish sampled during the two different outbreaks. Bacteriological, serological, molecular and chemotaxonomic analysis allowed the identification of the causative agent of sea bass mortalities as Aeromonas salmonicida subsp. salmonicida. The bacterial strains were susceptible to most of antimicrobial agents usually employed in aquaculture except to oxytetracycline. Pathogenicity assays demonstrated that the isolated bacteria were virulent for sea bass, turbot and rainbow trout.

Relation of chironomids with Aeromonas species

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Sivan eLaviad

2016-05-01

Full Text Available Chironomids (Diptera: Chironomidae, also known as non-biting midges, are one of the most abundant groups of insects in aquatic habitats. They undergo a complete metamorphosis of four life stages of which three are aquatic (egg, larva, pupa, and the adult emerges into the air. Chironomids serve as a natural reservoir of Aeromonas and Vibrio cholerae species. Here we review existing knowledge about the mutual relations between Aeromonas species and chironomids. Using 454-pyrosequencing of the 16S rRNA gene, we found that the prevalence of Aeromonas species in the insects’ egg masses and larvae was 1.6% and 3.3% of the insects’ endogenous microbiota, respectively. Aeromonas abundance per egg mass remained stable during a six-month period of bacterial monitoring. Different Aeromonas species were isolated and some demonstrated the ability to degrade the insect’s egg masses and to prevent eggs hatching. Chitinase was identified as the enzyme responsible for the egg mass degradation. Different Aeromonas species isolated from chironomids demonstrated the potential to protect their host from toxic metals. Aeromonas is a causative agent of fish infections. Fish are frequently recorded as feeding on chironomids. Thus, fish might be infected with Aeromonas species via chironomid consumption. Aeromonas strains are also responsible for causing gastroenteritis and wound infections in humans. Different virulence genes were identified in Aeromonas species isolated from chironomids. Chironomids may infest drinking water reservoirs, hence be the source of pathogenic Aeromonas strains in drinking water. Chironomids and Aeromonas species have a complicated mutual relationship.

Le cobaye Cavia porcellus L., comme animal de boucherie au Cameroun

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Ngou Ngoupayou, JD.

1994-01-01

Full Text Available Guinea Pig Cavia porcellus L. As A Meat Producing Animal In Cameroon. Guinea pig Cavia porcellus farming for meat production remains a marginalised activity in Cameroon in spite of the advantages this specie offers. With the view to promote its production, a national countrywide survey was carried out in order to evaluate the production systems and constraints. Traditional guinea pig farming appears to be a secondary household activity undertaken by small farmers basically women. The extensive production system which reveals no management practices, integrates very well in the agricultural systems (small livestock, food crops and natural forages production of the western highlands and southern forest zones of Cameroon. Guinea pig productivity remains low due to many constraints such as predation, uncontrolled breeding, inbreeding, poor feeding, negative selection and lack of veterinary care. Nevertheless, guinea pig farming plays an important role in the well being of low income village dwellers as food security, cash savings and socio-cultural values within the populations of South Cameroon. The promotion of its production requires an awareness of animal scientists, development authorities, as well as on station research if improved raising conditions.

Feasibility of same-day identification of members of the family Vibrionaceae by the API 20E system.

OpenAIRE

Overman, T L; Overley, J K

1986-01-01

Sixty isolates, comprising nine species of the family Vibrionaceae, were tested with the API 20E 5-h same-day procedure (Analytab Products, Plainview, N.Y.). Included were 27 Aeromonas hydrophila isolates, 10 Aeromonas sobria isolates, 7 Aeromonas caviae isolates, 3 Plesiomonas shigelloides isolates, 3 Vibrio alginolyticus isolates, 3 Vibrio cholerae isolates, 1 Vibrio fluvialis isolate, 5 Vibrio parahaemolyticus isolates, and 1 Vibrio vulnificus isolate. The 5-h profile numbers were specific...

Characterization of Aeromonas hydrophila wound pathotypes by comparative genomic and functional analyses of virulence genes.

Science.gov (United States)

Grim, Christopher J; Kozlova, Elena V; Sha, Jian; Fitts, Eric C; van Lier, Christina J; Kirtley, Michelle L; Joseph, Sandeep J; Read, Timothy D; Burd, Eileen M; Tall, Ben D; Joseph, Sam W; Horneman, Amy J; Chopra, Ashok K; Shak, Joshua R

2013-04-23

Aeromonas hydrophila has increasingly been implicated as a virulent and antibiotic-resistant etiologic agent in various human diseases. In a previously published case report, we described a subject with a polymicrobial wound infection that included a persistent and aggressive strain of A. hydrophila (E1), as well as a more antibiotic-resistant strain of A. hydrophila (E2). To better understand the differences between pathogenic and environmental strains of A. hydrophila, we conducted comparative genomic and functional analyses of virulence-associated genes of these two wound isolates (E1 and E2), the environmental type strain A. hydrophila ATCC 7966(T), and four other isolates belonging to A. aquariorum, A. veronii, A. salmonicida, and A. caviae. Full-genome sequencing of strains E1 and E2 revealed extensive differences between the two and strain ATCC 7966(T). The more persistent wound infection strain, E1, harbored coding sequences for a cytotoxic enterotoxin (Act), a type 3 secretion system (T3SS), flagella, hemolysins, and a homolog of exotoxin A found in Pseudomonas aeruginosa. Corresponding phenotypic analyses with A. hydrophila ATCC 7966(T) and SSU as reference strains demonstrated the functionality of these virulence genes, with strain E1 displaying enhanced swimming and swarming motility, lateral flagella on electron microscopy, the presence of T3SS effector AexU, and enhanced lethality in a mouse model of Aeromonas infection. By combining sequence-based analysis and functional assays, we characterized an A. hydrophila pathotype, exemplified by strain E1, that exhibited increased virulence in a mouse model of infection, likely because of encapsulation, enhanced motility, toxin secretion, and cellular toxicity. Aeromonas hydrophila is a common aquatic bacterium that has increasingly been implicated in serious human infections. While many determinants of virulence have been identified in Aeromonas, rapid identification of pathogenic versus nonpathogenic

Molecular detection of Aeromonas hydrophila in the aquarium gold fish and cultured rainbow trout in Chaharmahal va Bakhtiary province

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firouz Fadaeifard

2014-05-01

Full Text Available Aeromonas hydrophilia   is the etiologic agent of motile aeromonas septicaemia, one of the most important bacterial diseases of fresh and marine water fishes. The aim of the present study was detection of A. hydrophilia in the aquarium goldfish and cultured rainbow trout in Chaharmahal va Bakhtiary province.  In this study 50 goldfish from aquarium fish shops and 60 rainbow trouts suspected of having the disease from 6 farms (10 fish in each farm were randomly collected. The average weight in goldfish and rainbow trout samples were 3-5 g and 10-20 g, respectively. Sampling was performed from kidney and liver, and inoculated into blood agar and incubated at 22°C for 24 hours. Pure colonies which are grown on the mediums were tested by catalase, oxidase and gram staining, then those of gram-negative, catalase and oxidase positive were diagnosed, and cultured on Shotts-Rimler medium (as selective medium for A. hydrophila. These mediums were incubated at 22 °C for 24-48 h. The typical colonies were tested by using oligonucleotide primers of lip gene by PCR method. In light of molecular analysis of all specimens, 9 and 6 isolates from rainbow trout and gold fishes were identified as A. hydrophila respectively. Due to the detection of A. hydrophila in both cultured rainbow trout and aquarium goldfish, the bacteria can lead to septicemia with mortality if the health management principles are not observed in fish farming.

Novo meio seletivo-indicador para detecção de Aeromonas e Plesiomonas: ágar UNISC New selective indicator medium for detection of Aeromonas and Plesiomonas: UNISC agar

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Marion Pereira da Rocha

2008-08-01

Full Text Available Avaliou-se um novo meio seletivo-indicador (ágar UNISC para o isolamento de enteropatógenos clássicos e Aeromonas e Plesiomonas shigelloides. A capacidade de fermentação da xilose é indicada pela coloração amarela (fermentadores ou azul (não fermentadores que, aliada à prova da oxidase, constitui-se em indicador para a detecção de Aeromonas spp e Plesiomonas shigelloides. A produtividade e seletividade, avaliadas pelos índice de contagem absoluta e índice de contagem relativa indicam-no como uma alternativa aos coprocultivos clássicos porque permite, num só meio, o isolamento de Escherichia coli, Shigella spp, Salmonella spp, bem como, Aeromonas spp e Plesiomonas shigelloides, favorecendo o diagnóstico laboratorial das gastroenterites.We evaluated a new selective indicator medium (UNISC Agar for isolation of classical enteropathogens, Aeromonas spp and Plesiomonas shigelloides. The xylose fermentation capacity is indicated by a yellow color (fermenting agents or blue (no fermenting agent. This, together with the oxidase test, establishes it as an indicator for detecting Aeromonas and Plesiomonas shigelloides. Its productivity and selectivity, as assessed using the absolute count index and relative count index, indicate it as an alternative to the classical feces culturing media. This is because, in a single medium, it enables isolation of Escherichia coli, Shigella spp and Salmonella spp, in addition to Aeromonas and Plesiomonas shigelloides, thereby favoring the laboratory diagnosis of gastroenteritis.

CAViaR-based forecast for oil price risk

International Nuclear Information System (INIS)

Huang, Dashan; Yu, Baimin; Fabozzi, Frank J.; Fukushima, Masao

2009-01-01

As a benchmark for measuring market risk, value-at-risk (VaR) reduces the risk associated with any kind of asset to just a number (amount in terms of a currency), which can be well understood by regulators, board members, and other interested parties. This paper employs a new VaR approach due to Engle and Manganelli [Engle, R.F., Manganelli, S., 2004. CAViaR: Conditional Autoregressive Value at Risk by Regression Quantiles. Journal of Business and Economic Statistics 22, 367-381] to forecasting oil price risk. In doing so, we provide two original contributions by introducing a new exponentially weighted moving average CAViaR model and developing a mixed data regression model for multi-period VaR prediction. (author)

Molecular detection and antimicrobial resistance of Aeromonas from houseflies (Musca domestica in Iran

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Davood Ommi

2015-11-01

Full Text Available Objective. This study aimed to report the molecular detection and antimicrobial resistance of Aeromonas among houseflies (Musca domestica in Shahrekord and Isfahan provinces of Iran. Materials and methods. Flies were caught from household kitchens, cattle farms, animal hospitals, human hospitals, slaughter house and poultry farms and put in collection separate sterile tubes. Isolation was accomplished by culture of flies in alkaline peptone water followed by identification with Aeromonas-specific Polymerase Chain Reaction (PCR. Results. Out of 600 houseflies 73 (12.2% were infected with Aeromonas spp. Significantly higher frequencies of Aeromonas were isolated in Shahrekord province (13.0%; 39/300 than in Isfahan province (11.3%; 34/300. The recovery frequencies of the organisms were significantly lower in kitchens as compared to those in cattle farms and hospital wards which were similar. Higher proportions of infected flies were obtained during summer whereas low proportions were obtained during winter. Conclusions. It is concluded that houseflies do harbor diarrheagenic pathogens, including Aeromonas especially during summer. The carried organisms are resistant to a number of antimicrobials at different levels. Thus, future plans aimed at stemming infections caused by these organisms should take flies into account. Control efforts of infections caused by this particular bacterium should therefore take into account Musca domestica.

Molecular diversity among domestic guinea-pigs (Cavia porcellus and their close phylogenetic relationship with the Andean wild species Cavia tschudii Diversidad molecular entre cuyes domésticos (Cavia porcellus y su relación filogenética cercana con la especie silvestre andina Cavia tschudii

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ÁNGEL E SPOTORNO

2004-06-01

Full Text Available To investigate the origin and diversity of domestic guinea-pigs Cavia porcellus (Linnaeus, 1758; Rodentia, Caviidae, we sequenced the mitochondrial cytochrome b gene of 12 domestic and 10 wild specimens from six species, including the two presumed as ancestral to the domestic one: Cavia tschudii and Cavia aperea. All maximum parsimony and maximum likelihood analyses grouped C. porcellus with C. tschudii (mean K2P distance = 3.2 %; best trees had 609 steps (CI = 0.796; Bremer support Index (SI = 28, and a _Ln = 4419.52, with 100 % and 97 % bootstrap support respectively. This clade, supported by three substitutions and 96 % bootstrap, is also obtained in the cladistic analysis of corresponding amino acids. When the C. aperea node was forced to join C. porcellus, these trees were consistently longer, less likely and robust, and with less defining characters than the optimal one. All C. porcellus sequences also clustered in a node defined by 15 substitutions. The sub-node containing animals from city markets, pet shops and laboratories was characterized by four substitutions (one non-silent, SI = 7, and 91 % bootstrap. Some South American C. porcellus, called "criollos" (creoles by local breeders, were more diverse. Probably, a particular clade from southern Peru and Chile may represent a pre-Columbian lineage. Mean K2P distance between C. tschudii and C. aperea was rather large, 7.7 %. Cavia appeared as a robust node (100 % bootstrap. These results indicate that C. tschudii is the species most closely related to C. porcellusPara investigar el origen y la divergencia de los cuyes domésticos Cavia porcellus (Linnaeus, 1758; Rodentia, Caviidae, secuenciamos el gen mitocondrial para citocromo b de 12 especímenes domésticos y 10 silvestres de seis especies, incluyendo las dos que se presumen como ancestro de la doméstica: C. tschudii y C. aperea. Todos los análisis de máxima parsimonia y máxima verosimilitud agruparon a C. porcellus con C

Aeromonas veronii septicemia in an immunocompetent patient

OpenAIRE

Paula McKenzie; David Sotello; Yared Hailemariam; Vipul Desai; Brian Temple

2013-01-01

We present a 29-year-old healthy man who fell into an industrial auger, sustaining acrushed, open pelvic injury, multiple comminuted fractures of the right leg, and traumaticamputation of his left foot. Blood and wound cultures were positive for Aeromonasspp and vancomycin resistant Enterococcus. Treatment included cefepime, levofloxacin,daptomycin, and metronidazole. Aeromonas veronii is a Gram negative bacillus usuallyfound in fresh and brackish water in warm climates. It can cause severe s...

Sensitivity of planktonic and biofilm-associated Aeromonas spp. to ionizing radiation

International Nuclear Information System (INIS)

Nagar, Vandan; Bandekar, J.R.

2015-01-01

Genus Aeromonas has emerged as an important human pathogen because it causes a variety of diseases including gastroenteritis and extra-intestinal infections. Aeromonas have the ability to adhere and form biofilms on food surfaces and food contact surfaces. Biofilm formation on foods and food contact surfaces is the major reason for contamination, cross contamination and post-processing contamination of the final food product leading to food spoilage, product rejection, economic losses and food-borne diseases. Biofilms have shown high resistance to heat, desiccation, acidic condition, high salt concentration, antibiotics and other food preservatives. Earlier studies in our laboratory have shown that ionizing radiation effectively inactivates Aeromonas in different food products. However, the relative efficacy of this process against biofilm associated cells versus free-living planktonic cells of Aeromonas is not well documented. Therefore, the dose of gamma radiation required to reduce the population by 90% (D10) was calculated for planktonic and biofilm-associated A. salmonicida Y567 and A. hydrophila A331 cells. Both A. hydrophila A331 and A. salmonicida Y567 expressed significant ability to attach and grow on glass surface following incubation at 30℃ in TSB. Ionizing radiation effectively reduced the populations of both planktonic and biofilm-associated cells for both the strains. Mean cell counts of survivors and surviving fraction of planktonic and biofilm-associated cells decreased with increased irradiation doses. The D10 values of planktonic cells and biofilm cells for A. salmonicida (Y567) were 232.65 Gy and 248.41 Gy, respectively; whereas, the D10 values of planktonic cells and biofilm cells for A. hydrophila (A331) were 249.2 Gy and 240.2 Gy respectively. No significant difference in the D10 values of planktonic and biofilm associated Aeromonas was observed. The influence of the cultured state of the organism, i.e., planktonic versus biofilm associated

Use of a chiA probe for detection of chitinase genes in bacteria from the Chesapeake Bay

Digital Repository Service at National Institute of Oceanography (India)

Ramaiah, N.; Hill, R.T.; Chun, J.; Ravel, J.; Matte, M.H.; Straube, W.L.; Colwell, R.R.

PCR primers specific for the chiA gene were designed by alignment and selection of highly conserved regions of chiA sequences from Serratia marcescens, Alteromonas sp., Bacillus circulans and Aeromonas caviae. These primers were used to amplify a...

Characterization of Aeromonas hydrophila Wound Pathotypes by Comparative Genomic and Functional Analyses of Virulence Genes

Science.gov (United States)

Grim, Christopher J.; Kozlova, Elena V.; Sha, Jian; Fitts, Eric C.; van Lier, Christina J.; Kirtley, Michelle L.; Joseph, Sandeep J.; Read, Timothy D.; Burd, Eileen M.; Tall, Ben D.; Joseph, Sam W.; Horneman, Amy J.; Chopra, Ashok K.; Shak, Joshua R.

2013-01-01

ABSTRACT Aeromonas hydrophila has increasingly been implicated as a virulent and antibiotic-resistant etiologic agent in various human diseases. In a previously published case report, we described a subject with a polymicrobial wound infection that included a persistent and aggressive strain of A. hydrophila (E1), as well as a more antibiotic-resistant strain of A. hydrophila (E2). To better understand the differences between pathogenic and environmental strains of A. hydrophila, we conducted comparative genomic and functional analyses of virulence-associated genes of these two wound isolates (E1 and E2), the environmental type strain A. hydrophila ATCC 7966T, and four other isolates belonging to A. aquariorum, A. veronii, A. salmonicida, and A. caviae. Full-genome sequencing of strains E1 and E2 revealed extensive differences between the two and strain ATCC 7966T. The more persistent wound infection strain, E1, harbored coding sequences for a cytotoxic enterotoxin (Act), a type 3 secretion system (T3SS), flagella, hemolysins, and a homolog of exotoxin A found in Pseudomonas aeruginosa. Corresponding phenotypic analyses with A. hydrophila ATCC 7966T and SSU as reference strains demonstrated the functionality of these virulence genes, with strain E1 displaying enhanced swimming and swarming motility, lateral flagella on electron microscopy, the presence of T3SS effector AexU, and enhanced lethality in a mouse model of Aeromonas infection. By combining sequence-based analysis and functional assays, we characterized an A. hydrophila pathotype, exemplified by strain E1, that exhibited increased virulence in a mouse model of infection, likely because of encapsulation, enhanced motility, toxin secretion, and cellular toxicity. PMID:23611906

Multifocal Aeromonas Osteomyelitis in a Child with Leukemia

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Dimitrios Doganis

2016-01-01

Full Text Available Aeromonas hydrophila is a Gram negative organism causing both intestinal and extraintestinal disease. The case of a 14-year-old girl with underlying immunodeficiency and leukemia who developed systemic A. hydrophila infection is described in this report. While in deep bone marrow aplasia she developed fever, severe pain in the lower extremities, and swelling of the left femur. Blood culture showed Escherichia coli and A. hydrophila whereas pus culture from the soft tissue swelling showed the presence of A. hydrophila. Imaging studies showed diffuse osteolytic lesions. Patient received 5 months of intravenous and oral antibiotics and she improved clinically whereas the radiology findings persisted.

Plasmid-cured Chlamydia caviae activates TLR2-dependent signaling and retains virulence in the guinea pig model of genital tract infection.

Science.gov (United States)

Frazer, Lauren C; Darville, Toni; Chandra-Kuntal, Kumar; Andrews, Charles W; Zurenski, Matthew; Mintus, Margaret; AbdelRahman, Yasser M; Belland, Robert J; Ingalls, Robin R; O'Connell, Catherine M

2012-01-01

Loss of the conserved "cryptic" plasmid from C. trachomatis and C. muridarum is pleiotropic, resulting in reduced innate inflammatory activation via TLR2, glycogen accumulation and infectivity. The more genetically distant C. caviae GPIC is a natural pathogen of guinea pigs and induces upper genital tract pathology when inoculated intravaginally, modeling human disease. To examine the contribution of pCpGP1 to C. caviae pathogenesis, a cured derivative of GPIC, strain CC13, was derived and evaluated in vitro and in vivo. Transcriptional profiling of CC13 revealed only partial conservation of previously identified plasmid-responsive chromosomal loci (PRCL) in C. caviae. However, 2-deoxyglucose (2DG) treatment of GPIC and CC13 resulted in reduced transcription of all identified PRCL, including glgA, indicating the presence of a plasmid-independent glucose response in this species. In contrast to plasmid-cured C. muridarum and C. trachomatis, plasmid-cured C. caviae strain CC13 signaled via TLR2 in vitro and elicited cytokine production in vivo similar to wild-type C. caviae. Furthermore, inflammatory pathology induced by infection of guinea pigs with CC13 was similar to that induced by GPIC, although we observed more rapid resolution of CC13 infection in estrogen-treated guinea pigs. These data indicate that either the plasmid is not involved in expression or regulation of virulence in C. caviae or that redundant effectors prevent these phenotypic changes from being observed in C. caviae plasmid-cured strains.

Plasmid-cured Chlamydia caviae activates TLR2-dependent signaling and retains virulence in the guinea pig model of genital tract infection.

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Lauren C Frazer

Full Text Available Loss of the conserved "cryptic" plasmid from C. trachomatis and C. muridarum is pleiotropic, resulting in reduced innate inflammatory activation via TLR2, glycogen accumulation and infectivity. The more genetically distant C. caviae GPIC is a natural pathogen of guinea pigs and induces upper genital tract pathology when inoculated intravaginally, modeling human disease. To examine the contribution of pCpGP1 to C. caviae pathogenesis, a cured derivative of GPIC, strain CC13, was derived and evaluated in vitro and in vivo. Transcriptional profiling of CC13 revealed only partial conservation of previously identified plasmid-responsive chromosomal loci (PRCL in C. caviae. However, 2-deoxyglucose (2DG treatment of GPIC and CC13 resulted in reduced transcription of all identified PRCL, including glgA, indicating the presence of a plasmid-independent glucose response in this species. In contrast to plasmid-cured C. muridarum and C. trachomatis, plasmid-cured C. caviae strain CC13 signaled via TLR2 in vitro and elicited cytokine production in vivo similar to wild-type C. caviae. Furthermore, inflammatory pathology induced by infection of guinea pigs with CC13 was similar to that induced by GPIC, although we observed more rapid resolution of CC13 infection in estrogen-treated guinea pigs. These data indicate that either the plasmid is not involved in expression or regulation of virulence in C. caviae or that redundant effectors prevent these phenotypic changes from being observed in C. caviae plasmid-cured strains.

Incidence of Aeromonas spp. infection in fish and chicken meat and its related public health hazards: A review

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Praveen Kumar Praveen

2016-01-01

Full Text Available Aeromonas is recognized to cause a variety of diseases in man. In humans, they are associated with intestinal and extraintestinal infections. With the growing importance of Aeromonas as an emerging pathogen, it is important to combat this organism. It is indisputable that Aeromonas strains may produce many different putative virulence factors such as enterotoxins, hemolysins or cytotoxins, and antibiotic resistance against different antibiotics. The ability of these bacteria to grow competitively at 5°C may be indicative of their potential as a public health hazard. Comprehensive enteric disease surveillance strategies, prevention and education are essential for meeting the challenges in the years ahead. It is important for us to promote the value of enteric cultures when patients have a gastrointestinal illness or bloody diarrhea or when multiple cases of enteric disease occur after a common exposure. With the growing importance of Aeromonas as an emerging pathogen, it is important to combat this organism. It is indisputable that Aeromonas strains may produce many different putative virulence factors, such as enterotoxins, hemolysins or cytotoxins. It has been established that aerolysin is a virulence factor contributing to the pathogenesis of Aeromonas hydrophila infection. Fish and chicken play an important role in the transmission of this pathogen to humans. In the present study, the high prevalence of toxin-producing strains was found among the Aeromonas isolates. The ability of these bacteria to grow competitively at 5°C may be indicative of their potential as a public health hazard. The present review was constructed with a view to highlight the zoonotic importance of Aeromonas pathogen in fish and chicken meat.

Incidence of Aeromonas spp. infection in fish and chicken meat and its related public health hazards: A review.

Science.gov (United States)

Praveen, Praveen Kumar; Debnath, Chanchal; Shekhar, Shashank; Dalai, Nirupama; Ganguly, Subha

2016-01-01

Aeromonas is recognized to cause a variety of diseases in man. In humans, they are associated with intestinal and extra-intestinal infections. With the growing importance of Aeromonas as an emerging pathogen, it is important to combat this organism. It is indisputable that Aeromonas strains may produce many different putative virulence factors such as enterotoxins, hemolysins or cytotoxins, and antibiotic resistance against different antibiotics. The ability of these bacteria to grow competitively at 5°C may be indicative of their potential as a public health hazard. Comprehensive enteric disease surveillance strategies, prevention and education are essential for meeting the challenges in the years ahead. It is important for us to promote the value of enteric cultures when patients have a gastrointestinal illness or bloody diarrhea or when multiple cases of enteric disease occur after a common exposure. With the growing importance of Aeromonas as an emerging pathogen, it is important to combat this organism. It is indisputable that Aeromonas strains may produce many different putative virulence factors, such as enterotoxins, hemolysins or cytotoxins. It has been established that aerolysin is a virulence factor contributing to the pathogenesis of Aeromonas hydrophila infection. Fish and chicken play an important role in the transmission of this pathogen to humans. In the present study, the high prevalence of toxin-producing strains was found among the Aeromonas isolates. The ability of these bacteria to grow competitively at 5°C may be indicative of their potential as a public health hazard. The present review was constructed with a view to highlight the zoonotic importance of Aeromonas pathogen in fish and chicken meat.

The effect of infectious dose on humoral and cellular immune responses in Chlamydophila caviae primary ocular infection.

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Ana Filipovic

Full Text Available Following infection, the balance between protective immunity and immunopathology often depends on the initial infectious load. Several studies have investigated the effect of infectious dose; however, the mechanism by which infectious dose affects disease outcomes and the development of a protective immune response is not known. The aim of this study was to investigate how the infectious dose modulates the local and systemic humoral and the cellular immune responses during primary ocular chlamydial infection in the guinea pig animal model. Guinea pigs were infected by ocular instillation of a Chlamydophila caviae-containing eye solution in the conjunctival sac in three different doses: 1×102, 1×104, and 1×106 inclusion forming units (IFUs. Ocular pathology, chlamydial clearance, local and systemic C. caviae-specific humoral and cellular immune responses were assessed. All inocula of C. caviae significantly enhanced the local production of C. caviae-specific IgA in tears, but only guinea pigs infected with the higher doses showed significant changes in C. caviae-specific IgA levels in vaginal washes and serum. On complete resolution of infection, the low dose of C. caviae did not alter the ratio of CD4+ and CD8+ cells within guinea pigs' submandibular lymph node (SMLN lymphocytes while the higher doses increased the percentages of CD4+ and CD8+ cells within the SMLN lymphocytes. A significant negative correlation between pathology intensity and the percentage of CD4+ and CD8+ cells within SMLN lymphocyte pool at selected time points post-infection was recorded for both 1×104, and 1×106 IFU infected guinea pigs. The relevance of the observed dose-dependent differences on the immune response should be further investigated in repeated ocular chlamydial infections.

Treatment and control of Trixacarus caviae infestation in a conventional guinea pig (Cavia porcellus) breeding colony

OpenAIRE

Nath, Anjan Jyoti

2015-01-01

A case of sarcoptic mange caused by Trixacarus caviae in a conventional guinea pig breeding colony is reported. The infestation was reported in a large colony of guinea pigs during the month of July, 2013 affecting 30 breeder guinea pigs. Severely infested animals were treated individually with subcutaneous injection of ivermectin 1 % w/v (Neomec®) at the rate of 400 µg/kg body weight 10 days apart. Three doses of ivermectin were sufficient to eliminate the parasite which tested negative afte...

Detection and quantification of Aeromonas salmonicida in fish tissue by real-time PCR

DEFF Research Database (Denmark)

Bartkova, Simona; Kokotovic, Branco; Skall, H. F.

2017-01-01

Furunculosis, a septicaemic infection caused by the bacterium Aeromonas salmonicida subsp. salmonicida, currently causes problems in Danish seawater rainbow trout production. Detection has mainly been achieved by bacterial culture, but more rapid and sensitive methods are needed. A previously...

Antimicrobial resistance prevalence of Aeromonas hydrophila isolates from motile Aeromonas septicemia disease

Science.gov (United States)

Kusdarwati, R.; Rozi; Dinda, N. D.; Nurjanah, I.

2018-04-01

Fish suffer, from bacteria, fungi, virus and parasites or by physical ailments. Gurami (Osphronemus gouramy), nila (Oreochromis niloticus), carp (Cyprinus carpio), catfish (Clarias sp.) were the most reported infections caused by Aeromonas are bacterial hemorrhagic septicemia or Motile Aeromonas Septicemia (MAS). Antibiotics are drugs of natural or synthetic origin that have the capacity to kill or to inhibit the growth of micro-organisms included MAS. However, the use of antibiotics in the long term can cause negative impacts, among others, feared the occurrence of bacterial resistance in certain antibiotics. The results showed five of isolates were sensitive to antibiotics of chloramphenicol, gentamycin, oxytetracycline, cefradoxil and nalidixic acid but resistant to vancomycin colistin sulphate, rifampisin, cephalosporin and novobiocin.

Performances de production des cobayes (Cavia porcellus L.) en ...

African Journals Online (AJOL)

Production performance of guinea pigs (Cavia porcellus L.) in function of dietary protein level. ABSTRACT. In Cameroon, guinea pigs raising is still practiced in kitchens and on the ground and their food is ... females were previously crossed with 12 males for a period of 31 days according to a factorial design of three.

Genética poblacional de cobayas de Colombia, Cavia spp. (Rodentia: Caviidae con marcadores moleculares RAPD

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Héctor Aníbal Campos

2008-09-01

Full Text Available En el presente estudio, mostramos los primeros resultados moleculares de formas colombianas de Cavia. Claramente, la población silvestre de C. anolaimae fue genéticamente diferenciada de la forma doméstica, C. porcellus, tal como ha sido demostrado por otros autores utilizando resultados morfométricos, osteológicos y cariotípicos. Ambas especies mostraron un considerable nivel de diversidad genética, aunque el segundo taxon mostró niveles mayores de esta diversidad. Los niveles de heterogeneidad genética también fueron mayores entre las poblaciones de C. porcellus (F ST = 0.254 que entre las poblaciones de C. anolaimae (F ST = 0.118. Esos niveles significativos de heterogeneidad genética, y los consiguientes bajos niveles de flujo génico, fueron discutidos comparativamente con los resultados por otros autores analizando otros marcadores moleculares (citocromo-b mitocondrial. Los resultados aquí mostrados son coherentes con un complejo proceso de domesticación en Cavia porcellus.Population genetics of Colombian Guinea Pigs, Cavia spp. (Rodentia: Caviidae with RAPD molecular markers. The genus Cavia occurs in South America, mainly in grasslands.. We collected blood samples from 97 individuals in six field populations and analyzed them with RAPD molecular markers. One wild type (C. anolaimae was differentiated from the domestic form (C. porcellus, in agreement with other authors who used morphological, osteological and karyotipic results. Genetic diversity was considerable in both species, but higher in C. porcellus. The levels of genetic heterogeneity were also higher among the populations of C. porcellus (F ST = 0.254 than among the populations of C. anolaimae (F ST = 0.118. These significant levels of genetic heterogeneity, and the low levels of gene flow, were consistent with a complex domestication process for Cavia porcellus. Rev. Biol. Trop. 56 (3: 1481-1501. Epub 2008 September 30.

Multilocus genetics to reconstruct aeromonad evolution

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Roger Frédéric

2012-04-01

Full Text Available Abstract Background Aeromonas spp. are versatile bacteria that exhibit a wide variety of lifestyles. In an attempt to improve the understanding of human aeromonosis, we investigated whether clinical isolates displayed specific characteristics in terms of genetic diversity, population structure and mode of evolution among Aeromonas spp. A collection of 195 Aeromonas isolates from human, animal and environmental sources was therefore genotyped using multilocus sequence analysis (MLSA based on the dnaK, gltA, gyrB, radA, rpoB, tsf and zipA genes. Results The MLSA showed a high level of genetic diversity among the population, and multilocus-based phylogenetic analysis (MLPA revealed 3 major clades: the A. veronii, A. hydrophila and A. caviae clades, among the eleven clades detected. Lower genetic diversity was observed within the A. caviae clade as well as among clinical isolates compared to environmental isolates. Clonal complexes, each of which included a limited number of strains, mainly corresponded to host-associated subsclusters of strains, i.e., a fish-associated subset within A. salmonicida and 11 human-associated subsets, 9 of which included only disease-associated strains. The population structure was shown to be clonal, with modes of evolution that involved mutations in general and recombination events locally. Recombination was detected in 5 genes in the MLSA scheme and concerned approximately 50% of the STs. Therefore, these recombination events could explain the observed phylogenetic incongruities and low robustness. However, the MLPA globally confirmed the current systematics of the genus Aeromonas. Conclusions Evolution in the genus Aeromonas has resulted in exceptionally high genetic diversity. Emerging from this diversity, subsets of strains appeared to be host adapted and/or “disease specialized” while the A. caviae clade displayed an atypical tempo of evolution among aeromonads. Considering that A. salmonicida has been

Genospecies and virulence factors of Aeromonas species in ...

African Journals Online (AJOL)

Introduction: Aeromonads of medical importance have been reported from numerous clinical, food, and water sources, but identification of genospecies and virulence factors of Aeromonas species from countries in North Africa and the Middle East are few. Methods: In total 99 Aeromonas species isolates from different ...

Purification and characterization of the chitosanase from Aeromonas ...

African Journals Online (AJOL)

Purification and characterization of the chitosanase from Aeromonas sp. HG08. Y Sun, J Zhang, S Wang. Abstract. In this work, the characterization of a chitosanase-producing bacterium isolated from soil was reported and this strain was grouped under the genus Aeromonas by virtue of its morphological, physiological ...

NDM 1 Gene Carrying Gram negative Bacteria Isolated from Rats ...

African Journals Online (AJOL)

In this study, we screened 56 Gram negative bacteria comprising: 3 isolates of Enterobacter ludwigii, 30 Pseudomonas aeruginosa, 22 Proteus mirabilis, and 1 Aeromonas caviae isolated from oral cavity and rectum of rats captured from commercial poultry houses in Ibadan, Oyo State, Nigeria that were resistant to at least ...

NIGERIAN VETERINARY JOURNAL

African Journals Online (AJOL)

ADEYEYE

In this study, we screened 56 Gram negative bacteria comprising: 3 isolates of Enterobacter ludwigii, 30 Pseudomonas aeruginosa, 22. Proteus mirabilis, and 1 Aeromonas caviae isolated from oral cavity and rectum of rats captured from commercial poultry houses in Ibadan, Oyo State, Nigeria that were resistant to at least ...

Sanitary evaluation of domestic water supply facilities with storage tanks and detection of Aeromonas, enteric and related bacteria in domestic water facilities in Okinawa Prefecture of Japan.

Science.gov (United States)

Miyagi, Kazufumi; Sano, Kouichi; Hirai, Itaru

2017-08-01

To provide for temporary restrictions of the public water supply system, storage tanks are commonly installed in the domestic water systems of houses and apartment buildings in Okinawa Prefecture of Japan. To learn more about the sanitary condition and management of these water supply facilities with storage tanks (hereafter called "storage tank water systems") and the extent of bacterial contamination of water from these facilities, we investigated their usage and the existence of Aeromonas, enteric and related bacteria. Verbal interviews concerning the use and management of the storage tank water systems were carried out in each randomly sampled household. A total of 54 water samples were collected for bacteriological and physicochemical examinations. Conventional methods were used for total viable count, fecal coliforms, identification of bacteria such as Aeromonas, Enterobacteriaceae and non-fermentative Gram-negative rods (NF-GNR), and measurement of residual chlorine. On Aeromonas species, tests for putative virulence factor and an identification using 16S rRNA and rpoB genes were also performed. Water from the water storage systems was reported to be consumed directly without boiling in 22 of the 54 houses (40.7%). 31 of the sampled houses had installed water storage tanks of more than 1 cubic meter (m 3 ) per inhabitant, and in 21 of the sampled houses, the tank had never been cleaned. In all samples, the total viable count and fecal coliforms did not exceed quality levels prescribed by Japanese waterworks law. Although the quantity of bacteria detected was not high, 23 NF-GNR, 14 Enterobacteriaceae and 5 Aeromonas were isolated in 42.6%, 7.4% and 3.7% of samples respectively. One isolated A. hydrophila and four A. caviae possessed various putative virulence factors, especially A. hydrophila which had diverse putative pathogenic genes such as aer, hlyA, act, alt, ast, ser, and dam. Many bacteria were isolated when the concentration of residual chlorine

Population dynamics and antimicrobial susceptibility of Aeromonas spp. along a salinity gradient in an urban estuary in Northeastern Brazil.

Science.gov (United States)

Silva, Camila Magalhães; Evangelista-Barreto, Norma Suely; Vieira, Regine Helena Silva Dos Fernandes; Mendonça, Kamila Vieira; Sousa, Oscarina Viana de

2014-12-15

The main objective of this study was to quantify population and identify culturable species of Aeromonas in sediment and surface water collected along a salinity gradient in an urban estuary in Northeastern Brazil. Thirty sediment samples and 30 water samples were collected from 3 sampling locations (A, B and C) between October 2007 and April 2008. The Aeromonas count was 10-7050CFU/mL (A), 25-38,500CFU/mL (B) andwater samples, and ∼100-37,500CFU/g (A), 1200-43,500CFU/g (B) andantibiotics tested. Resistance to erythromycin was mostly plasmidial. In conclusion, due to pollution, the Cocó River is contaminated by pathogenic strains of Aeromonas spp. with a high incidence of antibacterial resistance, posing a serious risk to human health. Copyright © 2014 Elsevier Ltd. All rights reserved.

Interlaboratorium vergelijking van het onderzoek naar Aeromonas spp. in drinkwater (tweede onderzoek)

NARCIS (Netherlands)

Versteegh JFM; During M; Heisterkamp SH; Havelaar AH

1988-01-01

Door middel van kwantitatief onderzoek naar Aeromonas-bacterien in drie verschillende monsters water (De Bilt, De Meent en De Laak) werd een vergelijking gemaakt van de resultaten van Aeromonas onderzoek in 15 laboratoria in het kader van het deelproject: "Aeromonas in drinkwater".

Susceptibility of Aeromonas Hydophila Isolates to Antimicrobial Drugs

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Igor Stojanov

2010-05-01

Full Text Available Aeromonas hydrophila is a microorganism widely distributed in nature: in water, soil, food. It is also part of the normal bacterial flora of many animals. As an opportune microorganism it is a secondary biological agent that contributes to the occurrence of a fish disease and its deterioration. Frequently, its presence is an indication of bad zoohygiene and zootechnical conditions in fish ponds. Reduced quality and quantity of feed, mechanical injuries, parasitosis, seasonal oscillation in temperature present some of the factors that produce favorable conditions for bacterial proliferation of aeromonas in fish ponds, so clinical symptoms of the disease occur. Aeromonas is almost always present in clinical isolates and may be unjustly accused for bad health of fish. Antibiotic therapy is applied even when the clinical findings are clear, what certainly effects the susceptibility to chemotherapeutics. The subject of our work was bacteriological examination of the material obtained from the carps with the observed skin changes and the carps without these changes. Also, antimicrobial susceptibility of Aeromonas hydrophila was tested. The aim of this research was to determined the presence of Aeromonas hydrophilia in the carp ponds and to test antibiotic susceptibility. The material consisted of the samples from the fish ponds where the carps were with and without changed skin. The method the isolation of Aeromonas hydrophila was used. The diffusion disk technique was used for testing antibiotic susceptibility. The isolates were tested for their susceptibility to Florephenikol, Flumequine, Olaqindox and Oxitetracycline. The obtained results point that antimicrobial susceptibility was the same regardless of the origin of the samples, i.e. the resistance was the same for both groups of samples (the strains isolated from the fish with skin changes and the strains from fish without changes on skin. The strains were highly resistant: 35% were resistant to

Quorum Sensing Regulation in Aeromonas hydrophila

DEFF Research Database (Denmark)

Garde, Christian; Bjarnsholt, Thomas; Givskov, Michael

2010-01-01

We present detailed results on the C4-HSL-mediated quorum sensing (QS) regulatory system of the opportunistic Gram-negative bacterium Aeromonas hydrophila. This bacterium contains a particularly simple QS system that allows for a detailed modeling of kinetics. In a model system (i.e., the Escheri......We present detailed results on the C4-HSL-mediated quorum sensing (QS) regulatory system of the opportunistic Gram-negative bacterium Aeromonas hydrophila. This bacterium contains a particularly simple QS system that allows for a detailed modeling of kinetics. In a model system (i...

Quantitative Recovery of Aeromonas hydrophila from Nsukka Sewage

African Journals Online (AJOL)

Sewage samples were analyzed for the recovery of Aeromonas hydrophila. The pH of the samples ranged from 8.2 - 9.6 and the temperature from 20.0 – 28.7. Samples were enriched in alkaline peptone water medium (pH 8.4) before plating on different selective media. Media tested for the recovery of Aeromonas ...

IDENTIFIKASI NEMATODA PARASIT PADA SALURAN PENCERNAAN MARMUT (Cavia cobaya SEBAGAI SUMBER BELAJAR BIOLOGI PADA MATERI INVERTEBRATA

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Septia Nurhasanah

2014-05-01

Full Text Available The research was conducted in the Laboratory of Science Education, University of Muhammadiyah Metro. The research is a descriptive analysis, morphological observations were made by identifying the nematode parasite found that Trichostrongylus sp, Strongyloides sp, Trichuris sp, Trichuris sp. The results showed the prevalence and intensity values is the large number of guinea pigs to parasites divided by the number of guinea pigs that are 100% inspected. While the intensity is seen from the total number of nematode parasites that infect in the number of guinea pigs infected with a parasite that is 2,3. The population in this study was 10 cows pigs (Cavia cobaya . This study took a part in the digestion of marmots (Cavia cobaya the intestines and stomach , then cut and taken part in the large intestine and be Indian ink or coloring 2-3 drops to be easily identified microscopically . Put the preparations into a petri dish , and given alcohol as much as 96 % with 2-3 drops . Taking part in the examination of the large intestine to taste then used the nematode worm structure with Indian ink dye to be observed , and placed in the glass object . Observed with a microscope. Document if the parasites are found by using the camera Erlina 2008. Identification of the observations made have found 3 types of parasites that attack the guinea pig (Cavia cobaya is Strongyloides sp on samples 1,2,3,4,5,6 , dan10 . Then parasite Trichuris sp species found in samples 8 and 9. Trichostrongylus sp parasite species found in sample 7 . Parasitic nematodes found were 3 types of Trichostrongylus sp, Strongyloides sp, Trichuris sp. Number of samples that have been observed as many as 10 cows pigs (Cavia cobaya , with the overall result is as much as 23 tails parasites.               Kata kunci: nematode parasit, marmut (Cavia cobaya, sumber belajar biologi.

Evaluating virulence of waterborne and clinical Aeromonas isolates using gene expression and mortality in neonatal mice followed by assessing cell culture’s ability to predict virulence based on transcriptional response

Energy Technology Data Exchange (ETDEWEB)

Hayes, S L; Rodgers, M R; Lye, D J; Stelma, G N; McKinstry, Craig A.; Malard, Joel M.; Vesper, Sephen J.

2007-10-01

Aims: To assess the virulence of Aeromonas spp. using two models, a neonatal mouse assay and a mouse intestinal cell culture. Methods and Results: After artificial infection with a variety of Aeromonas spp., mRNA extracts from the two models were processed and hydridized to murine microarrays to determine host gene response. Definition of virulence was determined based on host mRNA production in murine neonatal intestinal tissue and mortality of infected animals. Infections of mouse intestinal cell cultures were then performed to determine whether this simpler model system’s mRNA responses correlated to neonatal results and therefore be predictive of virulence of Aeromonas spp. Virulent aeromonads up-regulated transcripts in both models including multiple host defense gene products (chemokines, regulation of transcription and apoptosis and cell signalling). Avirulent species exhibited little or no host response in neonates. Mortality results correlated well with both bacterial dose and average fold change of up-regulated transcripts in the neonatal mice. Conclusions: Cell culture results were less discriminating but showed promise as potentially being able to be predictive of virulence. Jun oncogene up-regulation in murine cell culture is potentially predictive of Aeromonas virulence. Significance and Impact of the Study: Having the ability to determine virulence of waterborne pathogens quickly would potentially assist public health officials to rapidly assess exposure risks.

Aeromonas spp. IN PRODUCTS OF ANIMAL ORIGIN AND DRINKING WATER OF GOIÂNIA-GO Aeromonas spp. EM PRODUTOS DE ORIGEM ANIMAL E EM ÁGUA DE CONSUMO DE GOIÂNIA - GO

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Albenones José de Mesquita

2007-09-01

Full Text Available

A total of 53 samples of products from animal origin and drinking water was examined for the presence of Aeromonas spp. Aeromonas occurred in 26.41% (14/53 of the samples. The raw products showed high frequency of contamination and relatively high count Aeromonas hydrophila was the dominating species, 57.5%, being followed of A. veroni (22.5%, A. sobria (17.5% and A. media (2.5%. None of the Aeromonas spp. was isolated from chlorinated or unchlorinated drinking water.

KEY-WORDS: Aeromonas; animal origin products; drinking water.

Um total de 53 amostras de produtos de origem animal e água de consumo foi examinado para se verificar a presença de Aeromonas spp. Desse total, 14/53 (26,41% amostras foram positivas para Aeromonas. Os produtos crus apresentaram maior freqüência de contaminação e contagem relativamente elevadas. A espécie Aeromonas hydrophila foi a mais freqüente (57,5%, seguida pelas espécies A. veroni (22,5%, A. sobria (17,5% e A. media (2,5%. Nenhuma amostra de água de consumo, clorada ou não clorada, mostrou-se positiva para Aeromonas ssp.

PALAVRA-CHAVE: Aeromonas; produtos de origem animal; água de consumo.

Prevalence of Aeromonas species and Escherichia coli in stool ...

African Journals Online (AJOL)

Background: Diarrhoea is one of the main causes of mortality and morbidity in childhood. Bacterial diarrhoea is a common disorder. Aeromonas species and Escherichia coli (E. coli) are some of the aetiological agents associated with diarrhoea in children. Objective: To determine the prevalence of Aeromonas species and ...

Do young guinea pigs (Cavia porcellus) develop an attachment to inanimate objects?

NARCIS (Netherlands)

Janzen, MID; Timmermans, PJA; Kruijt, JP; Vossen, JMH

1999-01-01

Filial imprinting has been studied extensively in precocial birds. In these studies, inanimate objects were used as imprinting objects. Although attachment to the parents is common in mammals, experiments with inanimate objects are rare and mostly restricted to guinea pigs (Cavia porcellus). The

Interlaboratorium vergelijking van het onderzoek naar Aeromonas spp. in drinkwater

NARCIS (Netherlands)

Havelaar AH; During M; Versteegh JFM

1986-01-01

Door middel van onderzoek van zes kunstmatig besmette gesimuleerde monsters drinkwater ( 4 Aeromonas spp., 1 Klebsiella oxytoca, 1 Pseudomonas aeruginosa) werd een vergelijking gemaakt van de resultaten van Aeromonas onderzoek in 14 laboratoria. De tellingen in de deelnemende laboratoria

Efecto modulador del sulfuro de hidrógeno en el metabolismo basal en Cavia porcellus (cuy) normales

OpenAIRE

Gómez Herreros, César Gonzalo

2016-01-01

Evalúa el efecto modulador del H2S a 80 ppm sobre el metabolismo basal en Cavia porcellus (cuy) normales. Para ello mide el consumo de oxígeno y producción de dióxido de carbono del Cavia porcellus (cuy) normales inmediatamente antes y después de la exposición al sulfuro de hidrógeno. Para luego determinar los cambios histológicos a nivel de tejido pulmonar, cardiaco y renal, y los niveles de hemoglobina entre los grupos experimentales (control, experimental y gold estándar). Tesis

The influence of p H during growth of bacteria in toluene

International Nuclear Information System (INIS)

Nahar, N.; Quilty, B.; Alauddin, M.

2000-01-01

Five toluene tolerant species were isolated from the activated sludge of a wastewater treatment plant (Dublin). The isolate were investigated for influence of pH on the growth in toluene. Four of the bacteria have been identified as Pseudomonas putida and one as an Aeromonas caviae. When these bacteria were grown with toluene as the sole source of carbon and energy, the pH of the culture medium became acidic and dropped. 0.5 M sodium phosphate buffer was selected to investigate the optimum pH for growth in the presence of 500 mu l of toluene. In general, the growth was optimum between pH 5.8 and 7.4. (author)

Experimental induction of motile Aeromonas septicemia in channel catfish (Ictalurus punctatus by waterborne challenge with virulent Aeromonas hydrophila

Directory of Open Access Journals (Sweden)

Dunhua Zhang

2016-05-01

Full Text Available Motile Aeromonas septicemia (MAS, caused by virulent clonal isolates of Aeromonas hydrophila (vAh, is emerging as a major disease in catfish (Ictalurus punctatus aquaculture in the Southeastern United States. Predisposing conditions leading to vAh infection in catfish were however largely unknown. The objective of this study was to investigate factors that predispose catfish to vAh infection and establish a waterborne challenge model that mimics natural occurrence of MAS. Results of this study indicated that wounding on the fish body surface was one of the key factors that predisposed catfish to vAh infection via waterborne route. Relatively uniform wounds were created by clipping part of the fish adipose fin. Adipose fin clipped (Af-clipped fish behaved normally in terms of swimming and feeding and no mortality occurred in the control treatment (a mock challenge. When subjected to challenge in vAh-infected water, Af-clipped fish were highly susceptible, showing typical symptoms of MAS observed in the field. The mortality rate of Af-clipped fish was significantly associated with vAh concentration, challenge time and water temperature. About 90% mortality occurred within 48 h when Af-clipped fish were challenged for 1 h with vAh at the concentration of 2 × 107 colony forming units per mL of water (27 ± 1 °C. The waterborne challenge model was further tested using four field isolates including A. hydrophila and A. veronii. All vAh isolates caused about 90% mortality of Af-clipped fish and one isolate of Aeromonas veronii caused no mortality under the same challenge conditions. The waterborne challenge model described in this study would facilitate urgently-needed studies of MAS prevention (such as wound avoidance and healing and control (such as prophylactic vaccination; antibiotics treatment and probiotics screening. Keywords: Adipose fin, Channel catfish, Motile Aeromonas septicemia, Virulent Aeromonas hydrophila, Waterborne challenge

Importancia biosanitaria de aeromonas: Taxonomia y Epidemiologia

OpenAIRE

Soler Falgàs, Lara

2003-01-01

El género Aeromonas está constituido por bacterias Gram-negativas aisladas frecuentemente en el agua. Adquieren relevancia por estar presentes en alimentos, y ser capaces de ocasionar cuadros diarreicos e infecciones extraintestinales en el hombre, habiéndose demostrado en la presente tesis, que pueden ser las causantes de hasta el 2% de las diarreas del viajero. La taxonomía de Aeromonas es compleja. El género consta de 15 especies difíciles de identificar con métodos bioquímicos, por lo cua...

Treatment and control of Trixacarus caviae infestation in a conventional guinea pig (Cavia porcellus) breeding colony.

Science.gov (United States)

Nath, Anjan Jyoti

2016-12-01

A case of sarcoptic mange caused by Trixacarus caviae in a conventional guinea pig breeding colony is reported. The infestation was reported in a large colony of guinea pigs during the month of July, 2013 affecting 30 breeder guinea pigs. Severely infested animals were treated individually with subcutaneous injection of ivermectin 1 % w/v (Neomec ® ) at the rate of 400 µg/kg body weight 10 days apart. Three doses of ivermectin were sufficient to eliminate the parasite which tested negative after 30 days of the first treatment. The entire colony was given preventive dose of ivermectin spray (2 mg/ml solution) following the same schedule. Strict hygienic measures were followed. New hair growth in the severely affected animals was evidenced on 30th day of treatment.

Co-operatives in the Cultural Industries

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Marisol Sandoval

2015-04-01

The roundtable discussed the potentials and limits of worker co-operatives as an alternative way of organizing cultural work. It explored how worker co-operation might contribute to new collaborative forms of cultural production; how they do, or might, strengthen a 'cultural commons'; and the role cultural co-ops play in the wider context of movements for workers' rights. Questions that were discussed include: To what extent can worker co-operatives be a means to confront precariousness and individualisation in work in the cultural sector? Do worker co-ops open up new possibilities for the collaborative production of cultural commons? What role can worker co-operatives play within a broader movement for creating more just, equal and humane cultural work and an alternative to capitalist economies? Where lies the boundary between neoliberal calls for self-help and individual responsibility and a radical co-op movement? What is the relation between worker co-ops and other forms of progressive politics such as the union movements, social protests and civil society activism? Can cultural co-ops contribute to reinventing the meaning and practice of work in the 21st century? About the speakers: Marisol Sandoval is a Lecturer at the Department of Culture and Creative Industries at City University London. Her research critically deals with questions of power, responsibility, commodification, exploitation, ideology and resistance in the global culture industry. Jo Littler is Senior Lecturer at City University London's Department of Culture and Creative Industries. Her work explores questions of culture and power from an interdisciplinary, cultural studies-informed perspective. Rhiannon Colvin after graduating in 2010 to find the world of work competitive and brutal, Rhiannon founded AltGen to empower young graduates to get together and create their own work. http://www.altgen.org.uk/ Tara Mulqueen is a PhD candidate at Birkbeck College School of Law. Her thesis concerns the

J-GLOBAL MeSH Dictionary: Aeromonas salmonicida [MeCab user dictionary for science technology term[Archive

Lifescience Database Archive (English)

Full Text Available MeCab user dictionary for science technology term Aeromonas salmonicida 名詞 一般 * * * * Aeromonas salmon...icida ... MeSH D048409 200906081596351600 C LS07 UNKNOWN_2 Aeromonas salmonicida

Antibacterial Effect of Gallic Acid against Aeromonas hydrophila and Aeromonas sobria Through Damaging Membrane Integrity.

Science.gov (United States)

Lu, Jing; Wang, Zhenning; Ren, Mengrou; Huang, Guoren; Fang, Baochen; Bu, Xiujuan; Liu, Yanhui; Guan, Shuang

In the study, we investigated the antibacterial activity and mechanism of gallic acid against Aeromonas hydrophila and Aeromonas sobria. Gallic acid showed strong antimicrobial activity against the two bacteria. Furthermore, the antibacterial mechanism of gallic acid (0, 3, 6, 12 mM) was performed by membrane integrity assay and scanning electron microscopy (SEM) assay. The results showed that gallic acid notably increased the released material absorption value at 260, 280 nm and electric conductivity in a dose-dependent manner. Moreover, the SEM assay showed that gallic acid induced severe shrink of bacterial intima and irregular morphology in a dose-dependent manner. The SDS-PAGE profiles further confirmed that gallic acid could damage bacterial cells. These results indicated gallic acid exhibited antibacterial effect by destroying membrane integrity of A. hydrophila and A. sobria. Hence, gallic acid has great potential as a new natural food preservative in food fresh-keeping and storage.

Een bijdrage tot de kennis van de experimentele tuberculeuze super-infectie bij de cavia

NARCIS (Netherlands)

Goslinga, Jan

1960-01-01

Er werd een beschrijving gegeven van een in 1954 in het sanatorium "de KLOKKENBERG" te Breda verricht onderzoek, waarbij 37 caviae achtereenvolgens enkele malen met virulente, tegen bepaalde stoffen selectief resistent gekweekte tuberkelbacteriën, afgeleid van &n bepaalde, bij een van de patienten

Aeromonas presence in drinking water from collective reservoirs and wells in peri-urban area in Brazil

Directory of Open Access Journals (Sweden)

Maria Tereza Pepe Razzolini

2010-10-01

Full Text Available Aeromonas genus is considered an emerging pathogen and its presence in drinking water supplies is a reason to public health concern. This study investigated the occurrence of Aeromonas in samples from collective reservoirs and wells used as drinking water sources in a peri-urban area. A total of 35 water samples were collected from collective reservoirs and 32 from wells bimonthly, from September 2007 to September 2008. Aeromonas spp determination was carried out using a Multiple-Tube Technique. Samples were inoculated into alkaline peptone water and the superficial film formed was transferred to blood agar plates amended with ampicillin. Typical Aeromonas colonies were submitted to a biochemical screening and then to biochemical tests for species differentiation. Aeromonas was detected in 13 (19% of the 69 samples examined (6 from collective reservoirs and 7 from wells. Concentrations of Aeromonas in collective reservoirs ranged from <0.3 to 1.2 x10²MPN/100mL and, in wells, from <0.3 to 2.4 x10²MPN/100mL. The most frequent specie in the collective reservoir samples was Aeromonas spp (68%, followed by A. encheleia (14% and A. allosaccharophila (8% and A. hydrophila (8%. Aeromonas spp (87% was the most frequent specie isolated from well samples, followed by A. allosacchariphila (8%, A. encheleia (2% and A. jandaei (5%. These data show the presence and diversity of Aeromonas genus in the samples analyzed and highlight that its presence in drinking water poses a significant public health concern.

Aminoglycoside-resistant Aeromonas hydrophila as part of a polymicrobial infection following a traumatic fall into freshwater.

Science.gov (United States)

Shak, Joshua R; Whitaker, Jennifer A; Ribner, Bruce S; Burd, Eileen M

2011-03-01

Amikacin is a first-line treatment for Aeromonas infection due to high efficacy. There are few reports of aminoglycoside-resistant Aeromonas spp. We report a soft tissue infection containing multiple pathogens, including a strain of Aeromonas hydrophila resistant to amikacin, tobramycin, and multiple cephalosporins.

Aminoglycoside-Resistant Aeromonas hydrophila as Part of a Polymicrobial Infection following a Traumatic Fall into Freshwater▿

Science.gov (United States)

Shak, Joshua R.; Whitaker, Jennifer A.; Ribner, Bruce S.; Burd, Eileen M.

2011-01-01

Amikacin is a first-line treatment for Aeromonas infection due to high efficacy. There are few reports of aminoglycoside-resistant Aeromonas spp. We report a soft tissue infection containing multiple pathogens, including a strain of Aeromonas hydrophila resistant to amikacin, tobramycin, and multiple cephalosporins. PMID:21209173

Prevalence, species differentiation, haemolytic activity, and antibiotic susceptibility of aeromonads in untreated well water

Directory of Open Access Journals (Sweden)

Khalifa Sifaw Ghenghesh

2001-02-01

Full Text Available The use of untreated water for drinking and other activities have been associated with intestinal and extraintestinal infections in humans due to Aeromonas species. In the present study aeromonads were isolated from 48.7% of 1,000 water samples obtained from wells and other miscellaneous sources. Aeromonas species were detected in 45% of samples tested in spring, 34.5% in summer, 48% in autumn and 60% of samples tested in winter. Speciation of 382 strains resulted in 225 (59% being A. hydrophila, 103 (27% A. caviae, 42 (11% A. sobria and 11 (3% atypical aeromonads. Of 171 Aeromonas strains tested for their haemolytic activity, 53%, 49%, 40% and 37% were positive in this assay using human, horse, sheep and camel erythrocytes respectively. The results obtained indicate that potentially enteropathogenic Aeromonas species are commonly present in untreated drinking water obtained from wells in Libya (this may also apply to other neighbouring countries which may pose a health problem to users of such water supplies. In addition, ceftriaxone and ciprofloxacin are suitable drugs that can be used in the treatment of Aeromonas-associated infections, particularly in the immunocompromised, resulting from contact with untreated sources of water.

Investigation Into the Humaneness of Slaughter Methods for Guinea Pigs (Cavia porcelus) in the Andean Region.

Science.gov (United States)

Limon, Georgina; Gonzales-Gustavson, Eloy A; Gibson, Troy J

2016-01-01

Guinea pigs (Cavia porcelus) are an important source of nonhuman animal protein in the Andean region of South America. Specific guidelines regarding the welfare of guinea pigs before and during slaughter have yet to be developed. This study critically assessed the humaneness of 4 different stunning/slaughter methods for guinea pigs: cervical neck dislocation (n = 60), electrical head-only stunning (n = 83), carbon dioxide (CO2) stunning (n = 21), and penetrating captive bolt (n = 10). Following cervical neck dislocation, 97% of guinea pigs had at least 1 behavioral or cranial/spinal response. Six percent of guinea pigs were classified as mis-stunned after electrical stunning, and 1% were classified as mis-stunned after captive bolt. Increased respiratory effort was observed during CO2 stunning. Apart from this finding, there were no other obvious behavioral responses that could be associated with suffering. Of the methods assessed, captive bolt was deemed the most humane, effective, and practical method of stunning guinea pigs. Cervical neck dislocation should not be recommended as a slaughter method for guinea pigs.

Aeromonas hydrophila urinary tract infection in pregnancy- Case report and literature review

OpenAIRE

Ragunathan, Latha; Kavitha, Kannaiyan; Raveendran, Vinod; Dhandapani, Senthil Pragash; Jaget, Nirmala; Kannivelu, Jayanthi

2012-01-01

A case of a pregnant woman without previous or concomitant disease, who developed an Aeromonas hydrophila urinary tract infection (UTI) at 12 weeks gestation, is reported. A brief review of the literature on the association and incidence of Aeromonas spp in urinary infections and also in association with pregnancy is presented.

Injury Thresholds for Topical Cream-Coated Skin of Hairless Guinea Pigs (Cavia Procellus) in Near Infrared Region

National Research Council Canada - National Science Library

Stolarski, David J; Zohner, Justin J; Imholte, Michelle L; Chavey, Lucas J; Pocock, Ginger M; Buchanan, Kelvin C; Jindra, Nichole M; Figueroa, Manuel A; Thomas, Robert J; Rockwell, Benjamin A

2006-01-01

.... Hairless guinea pigs (cavia porcellus) in vivo were used to evaluate how the minimum visible lesion threshold for single-pulse laser exposure is changed with a topical agent applied to the skin...

Antibiotic Susceptibility Profile of Aeromonas Species Isolated from Wastewater Treatment Plant

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Isoken H. Igbinosa

2012-01-01

Full Text Available This study assessed the prevalence of antibiotic-resistant Aeromonas species isolated from Alice and Fort Beaufort wastewater treatment plant in the Eastern Cape Province of South Africa. Antibiotic susceptibility was determined using the disc diffusion method, and polymerase chain reaction (PCR assay was employed for the detection of antibiotics resistance genes. Variable susceptibilities were observed against ciprofloxacin, chloramphenicol, nalidixic acid, gentamicin, minocycline, among others. Aeromonas isolates from both locations were 100% resistant to penicillin, oxacillin, ampicillin, and vancomycin. Higher phenotypic resistance was observed in isolates from Fort Beaufort compared to isolates from Alice. Class A pse1 β-lactamase was detected in 20.8% of the isolates with a lower detection rate of 8.3% for blaTEM gene. Class 1 integron was present in 20.8% of Aeromonas isolates while class 2 integron and TetC gene were not detected in any isolate. The antibiotic resistance phenotypes observed in the isolates and the presence of β-lactamases genes detected in some isolates are of clinical and public health concern as this has consequences for antimicrobial chemotherapy of infections associated with Aeromonas species. This study further supports wastewater as potential reservoirs of antibiotic resistance determinants in the environment.

Effects of Co-Culture Media on Hepatic Differentiation of hiPSC with or without HUVEC Co-Culture.

Science.gov (United States)

Freyer, Nora; Greuel, Selina; Knöspel, Fanny; Strahl, Nadja; Amini, Leila; Jacobs, Frank; Monshouwer, Mario; Zeilinger, Katrin

2017-08-07

The derivation of hepatocytes from human induced pluripotent stem cells (hiPSC) is of great interest for applications in pharmacological research. However, full maturation of hiPSC-derived hepatocytes has not yet been achieved in vitro. To improve hepatic differentiation, co-cultivation of hiPSC with human umbilical vein endothelial cells (HUVEC) during hepatic differentiation was investigated in this study. In the first step, different culture media variations based on hepatocyte culture medium (HCM) were tested in HUVEC mono-cultures to establish a suitable culture medium for co-culture experiments. Based on the results, two media variants were selected to differentiate hiPSC-derived definitive endodermal (DE) cells into mature hepatocytes with or without HUVEC addition. DE cells differentiated in mono-cultures in the presence of those media variants showed a significant increase ( p < 0.05) in secretion of α-fetoprotein and in activities of cytochrome P450 (CYP) isoenzymes CYP2B6 and CYP3A4 as compared with cells differentiated in unmodified HCM used as control. Co-cultivation with HUVEC did not further improve the differentiation outcome. Thus, it can be concluded that the effect of the used medium outweighed the effect of HUVEC co-culture, emphasizing the importance of the culture medium composition for hiPSC differentiation.

Aeromonas hydrophila urinary tract infection in pregnancy- Case report and literature review

Directory of Open Access Journals (Sweden)

Latha Ragunathan

2012-03-01

Full Text Available A case of a pregnant woman without previous or concomitant disease, who developed an Aeromonas hydrophila urinarytract infection (UTI at 12 weeks gestation, is reported. A brief review of the literature on the association and incidenceof Aeromonas spp in urinary infections and also in association with pregnancy is presented. J Microbiol Infect Dis 2012;2(1: 26-28

Aeromonas salmonicida bacteremia associated with chronic well water consumption in a patient with diabetes

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Christine Ann Moore

2017-01-01

Full Text Available Aeromonas salmonicida is associated with superficial skin infections in fish. Its virulence factors allow colonization of water including surface water such as salt water, beaches, and fresh water wells. Moreover, it is possible for immunocompromised patients to develop invasive disease after chronic exposure to Aeromonas spp. through contaminated water. While there are reports of Aeromonas spp. bacteremia following water ingestion, there have been no reports of A. salmonicida bacteremia from water consumption. We report the first case of A. salmonicida bacteremia in a patient with diabetes due to chronic consumption of well water.

AEROMONAS SALMONICIDA INFECTION IN VACCINATED RAINBOW TROUT

DEFF Research Database (Denmark)

Chettri, Jiwan Kumar; Skov, Jakob; Mohammad, Rezkar Jaafar

In vivo testing of any candidate vaccine is influenced by the choice of challenge method and the external environmental conditions. In the present study, a comparative challenge study was performed to evaluate the efficacy of different vaccines against the bacterial pathogen Aeromonas salmonicida...

[Real-time PCR in rapid diagnosis of Aeromonas hydrophila necrotizing soft tissue infections].

Science.gov (United States)

Kohayagawa, Yoshitaka; Izumi, Yoko; Ushita, Misuzu; Niinou, Norio; Koshizaki, Masayuki; Yamamori, Yuji; Kaneko, Sakae; Fukushima, Hiroshi

2009-11-01

We report a case of rapidly progressive necrotizing soft tissue infection and sepsis followed by a patient's death. We suspected Vibrio vulnificus infection because the patient's underlying disease was cirrhosis and the course extremely rapid. No microbe had been detected at death. We extracted DNA from a blood culture bottle. SYBR green I real-time PCR was conducted but could not detect V. vulnificus vvh in the DNA sample. Aeromonas hydrophila was cultured and identified in blood and necrotized tissue samples. Real-time PCR was conducted to detect A. hydrophila ahh1, AHCYTOEN and aerA in the DNA sample extracted from the blood culture bottle and an isolated necrotized tissue strain, but only ahh1 was positive. High-mortality in necrotizing soft tissue infections makes it is crucial to quickly detect V. vulnificus and A. hydrophila. We found real-time PCR for vvh, ahh1, AHCYTOEN, and aerA useful in detecting V. vulnificus and A. hydrophila in necrotizing soft tissue infections.

Evolutionary Roots and Diversification of the Genus Aeromonas.

Science.gov (United States)

Sanglas, Ariadna; Albarral, Vicenta; Farfán, Maribel; Lorén, J G; Fusté, M C

2017-01-01

Despite the importance of diversification rates in the study of prokaryote evolution, they have not been quantitatively assessed for the majority of microorganism taxa. The investigation of evolutionary patterns in prokaryotes constitutes a challenge due to a very scarce fossil record, limited morphological differentiation and frequently complex taxonomic relationships, which make even species recognition difficult. Although the speciation models and speciation rates in eukaryotes have traditionally been established by analyzing the fossil record data, this is frequently incomplete, and not always available. More recently, several methods based on molecular sequence data have been developed to estimate speciation and extinction rates from phylogenies reconstructed from contemporary taxa. In this work, we determined the divergence time and temporal diversification of the genus Aeromonas by applying these methods widely used with eukaryotic taxa. Our analysis involved 150 Aeromonas strains using the concatenated sequences of two housekeeping genes (approximately 2,000 bp). Dating and diversification model analyses were performed using two different approaches: obtaining the consensus sequence from the concatenated sequences corresponding to all the strains belonging to the same species, or generating the species tree from multiple alignments of each gene. We used BEAST to perform a Bayesian analysis to estimate both the phylogeny and the divergence times. A global molecular clock cannot be assumed for any gene. From the chronograms obtained, we carried out a diversification analysis using several approaches. The results suggest that the genus Aeromonas began to diverge approximately 250 millions of years (Ma) ago. All methods used to determine Aeromonas diversification gave similar results, suggesting that the speciation process in this bacterial genus followed a rate-constant (Yule) diversification model, although there is a small probability that a slight

Aquatically acquired Aeromonas hydrophila wound infection A ...

African Journals Online (AJOL)

The occurrence of Aeromonas hydrophila wound infections in healthy hosts after water-associated injury is being reported more frequently. This paper reports our experience with 3 such cases and outlines the importance of recognising the association between a water-related injury and, this organism.

Aeromonas spp. and microbial indicators in raw drinking water source Aeromonas sp. e indicadores microbiológicos em fontes de água não tratada

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Marisa Di Bari

2007-09-01

Full Text Available Aeromonas species are autochtonous in the aquatic environment and some of them have been associated with health effects like wound infections, septicemia and diarrhoeal illness. In this study, the occurrence of Aeromonas spp. and microbial indicators in raw drinking water from wells, springs, fountains and mineral waters was evaluated. A total of 126 water samples was analyzed for Aeromonas spp. by the membrane filtration technique using ADA media and by P/A test. Typical colonies of Aeromonas spp. were submitted to biochemical tests for species differentiation. Toxin production was tested using Y-1 mouse adrenal cells. Coliforms and heterotrophic bacteria were enumerated by membrane filtration and pour plate techniques, respectively. P. aeruginosa, C. perfringens and fecal streptococci were determined by P/A method. Aeromonas spp. were isolated in 36.5% of the samples, whereas total and thermotolerant coliforms were detected in 51.2% and in 23.8% of the samples, respectively. C. perfringens, fecal streptococci and P. aeruginosa were present in 16.5%, 20.4% and 3.8% of the samples, respectively. The concentrations of heterotrophic bacteria were higher than 1,0x10³ CFU/mL in 52.5% of the samples. A. hydrophila was the most frequent species, followed by A. allosaccharophila,A. jandaei,A.sobria and HG2. A heat label toxin was detected in 13 from the 58 strains tested. These data show that the drinking water sources analyzed can represent a risk for human health. It is important to consider that wells and springs are used as drinking water supply in poor areas and rural regions, where undernourished people more susceptible to infections by these microorganisms predominate.Bactérias do gênero Aeromonas são naturais no ambiente aquático e algumas espécies podem causar infecções em humanos como feridas, septicemia e diarréia. Este trabalho objetivou avaliar a ocorrência de Aeromonas sp. em 126 amostras de água de poços, nascentes, fontes e

A comparative analysis of the intestinal metagenomes present in guinea pigs (Cavia porcellus) and humans (Homo sapiens)

DEFF Research Database (Denmark)

Hildebrand, Falk; Ebersbach, Tine; Nielsen, Henrik Bjørn

2012-01-01

Background: Guinea pig (Cavia porcellus) is an important model for human intestinal research. We have characterized the faecal microbiota of 60 guinea pigs using Illumina shotgun metagenomics, and used this data to compile a gene catalogue of its prevalent microbiota. Subsequently, we compared th...

Identification of Two Pathogenic Aeromonas Species Isolated during Production Related Epizootics from Juvenile Burbot Lota lota maculosa.

Science.gov (United States)

Terrazas, Marc M; Anderson, Cort L; Jacobs, Sarah J; Cain, Kenneth D

2018-05-25

In response to population declines of Burbot Lota lota maculosa, conservation aquaculture methods have been developed for this species. In general, Burbot are relatively resistant to many salmonid pathogens; however, cultured juvenile Burbot have experienced periodic epizootic disease outbreaks during production. A series of trials were conducted to determine the virulence of select bacteria isolated from juvenile Burbot following outbreaks that occurred in 2012 and 2013 at the University of Idaho's Aquaculture Research Institute. Initial clinical diagnostics and sampling resulted in the isolation of numerous putative bacterial pathogens. To determine which bacteria were the most likely causative agents contributing to these epizootics, juvenile Burbot were injectied intraperitoneally (IP) with select bacteria in log phase growth. Mortality associated with specific isolates was recorded, and more comprehensive challenges followed this initial screening. These challenges used side-by-side IP and immersion methods to expose Burbot to potential pathogens. The challenges resulted in significantly higher mortalities in fish following IP injection of two Aeromonas sp. isolates compared to controls (P ≤ 0.01), but no difference in mortality for immersion challenged groups was observed (P = 0.42). Results demonstrate that two isolates (Aeromonas sp.) cultured from the epizootics are virulent to Burbot. This article is protected by copyright. All rights reserved. This article is protected by copyright. All rights reserved.

A foodborne outbreak of Aeromonas hydrophila in a college, Xingyi City, Guizhou, China, 2012.

Science.gov (United States)

Zhang, Qian; Shi, Guo-Qing; Tang, Guang-Peng; Zou, Zhi-Tin; Yao, Guang-Hai; Zeng, Guang

2012-10-01

On 12 May 2012, over 200 college students with acute diarrhoea were reported to the Guizhou Center for Disease Control and Prevention. We conducted an investigation to identify the agent and mode of transmission and to recommend control measures. A suspected case was a person at the college with onset of ≥ two of the following symptoms: diarrhoea (more than three loose stools in 24 hours), abdominal pain, vomiting or fever (> 37.5C) between 6 and 15 May 2012. A confirmed case also had a positive Aeromonas hydrophila culture from a stool sample. A retrospective-cohort study of 902 students compared attack rates (AR) by dining place, meals and food history. We reviewed the implicated premise, its processes and preparation of implicated food. We identified 349 suspected cases (AR = 14%) and isolated Aeromonas hydrophila from three stools of 15 cases. Students who ate in cafeteria A were more likely to be ill compared to those eating in other places (relative risk [RR]: 3.1, 95% confidence interval [CI]: 2.0-4.8). The cohort study implicated cold cucumber (RR: 2.6, 95% CI: 2.0-3.3) and houttuynia dishes (RR: 1.8, 95% CI: 1.4-2.3). Environmental investigation showed that vegetables were washed in polluted water from a tank close to the sewage ditch, then left at 30 °C for two hours before serving. The Escherichia coli count of the tank was well above the standard for drinking-water. This outbreak of Aeromonas hydrophila was most probably caused by salad ingredients washed in contaminated tank water. We recommended enhancing training of foodhandlers, ensuring tanks and sewerage systems comply with appropriate standards and adequate monitoring of drinking-water sources.

A foodborne outbreak of Aeromonas hydrophila in a college, Xingyi City, Guizhou, China, 2012

Directory of Open Access Journals (Sweden)

Guang Zeng

2012-12-01

Full Text Available Background: On 12 May 2012, over 200 college students with acute diarrhoea were reported to the Guizhou Center for Disease Control and Prevention. We conducted an investigation to identify the agent and mode of transmission and to recommend control measures.Methods: A suspected case was a person at the college with onset of more than two of the following symptoms: diarrhoea (more than three loose stools in 24 hours, abdominal pain, vomiting or fever (> 37.5C between 6 and 15 May 2012. A confirmed case also had a positive Aeromonas hydrophila culture from a stool sample. A retrospective-cohort study of 902 students compared attack rates (AR by dining place, meals and food history. We reviewed the implicated premise, its processes and preparation of implicated food.Results: We identified 349 suspected cases (AR = 14% and isolated Aeromonas hydrophila from three stools of 15 cases. Students who ate in cafeteria A were more likely to be ill compared to those eating in other places (relative risk [RR]: 3.1, 95% confidence interval [CI]: 2.0–4.8. The cohort study implicated cold cucumber (RR: 2.6, 95% CI: 2.0–3.3 and houttuynia dishes (RR: 1.8, 95% CI: 1.4–2.3. Environmental investigation showed that vegetables were washed in polluted water from a tank close to the sewage ditch, then left at 30°C for two hours before serving. The Escherichia coli count of the tank was well above the standard for drinking water.Conclusion: This outbreak of Aeromonas hydrophila was most probably caused by salad ingredients washed in contaminated tank water. We recommended enhancing training of foodhandlers, ensuring tanks and sewerage systems comply with appropriate standards and adequate monitoring of drinking water sources.

Isolation and Identification of Aeromonas hydrophila and Saprolegnia sp. on Catfish (Clarias gariepinus) in Floating cages in Bozem Moro Krembangan Surabaya

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Kusdarwati, Rahayu; Kismiyati; Sudarno; Kurniawan, Hendi; Teguh Prayogi, Yudha

2017-02-01

Catfish (Clarias gariepinus) is one of the familiar freshwater fish cultured in Indonesia farmer. One of the reason is the high mortality of the catfish infected by Aeromonas hydrophila and Saprolegnia sp. Motile Aeromonas septicemia (MAS) is a common bacterial disease, caused by Aeromonas hydrophila, which affects freshwater fish. In Southeast Asia, the outbreak of this disease was firstly reported from West Java in 1980, when a total of 82.5 tons a month of catfish were lost, while in Central Java in 1984, the total loss was 1.6 tons. Saprolegniosis can cause economic loss due to high mortality from its case reaching 10% to 50%. This research aimed to identify and determine the percentage of A. hydrophila and Saprolegnia sp. the catfish farmed in bozem Moro Krembangan, Surabaya, East Java. Meanwhile, a supporting parameter in this research is the value of water quality parameter including pH, temperature, ammonia and dissolved oxygen that were measured during sampling. The results showed that of the 20 samples taken from the two cages, 19 fish were positively infected by A. drophila. percentage of infections of A. hydrophila that infect umbo catfish in Moro Krembangan, was 95%, while the percentage Saprolegnia sp. was 90%.

Antimicrobial susceptibility pattern of Aeromonas hydrophila among ...

African Journals Online (AJOL)

A total of one hundred and twenty eight (128) stool samples were collected from patients attending Ahmadu Bello University Teaching Hospital Zaria and Aminu Kano Teaching Hospital Kano, and screened for the presence of Aeromonas hydrophila infection. Out of the one hundred and twenty eight samples analyzed 4 ...

Detection of Aeromonas hydrophila Using Fiber Optic Microchannel Sensor

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Samla Gauri

2017-01-01

Full Text Available This research focuses on the detection of Aeromonas hydrophila using fiber optic microchannel biosensor. Microchannel was fabricated by photolithography method. The fiber optic was chosen as signal transmitting medium and light absorption characteristic of different microorganisms was investigated for possible detection. Experimental results showed that Aeromonas hydrophila can be detected at the region of UV-Vis spectra between 352 nm and 354 nm which was comparable to measurement provided by UV spectrophotometer and also theoretical calculation by Beer-Lambert Absorption Law. The entire detection can be done in less than 10 minutes using a total volume of 3 μL only. This result promises good potential of this fiber optic microchannel sensor as a reliable, portable, and disposable sensor.

Cultural Issues of Co-Sleeping in Korea

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Seockhoon Chung

2014-12-01

Full Text Available Co-sleeping is a natural part of parenting in the Eastern culture; however, it may seem strange and possibly even dangerous to Western cultures. In the West, parental age, race, marital status, and house income may influence co-sleeping, while co-sleeping, especially bed-sharing, is usually considered to increase the risk of sudden infant death syndrome. In Korea, however, people usually believe that a baby must not sleep alone in an empty room. The differences in the prevalence of co-sleeping between Eastern and Western society may be rooted in differences in child-care philosophies, sleeping habits, and home architecture. In this article, the hazards and benefits of bed-sharing will be reviewed, and differences in co-sleeping will be addressed from a cultural viewpoint.

Bacteremia due to Aeromonas hydrophila in Acute Lymphoblastic Leukemia

International Nuclear Information System (INIS)

Fatima, A.; Afridi, F.I.; Farooqi, B.J.; Qureshi, A.; Hussain, A.

2013-01-01

Aeromonas hydrophila (A. hydrophila) is a low virulent organism but may cause devastating fatal infections in immunocompromised host especially in liver cirrhosis. It is rarely reported to cause septicemia in a patient with Acute Lymphoblastic Leukemia (ALL). The mortality rate of septicemia due to A. hydrophila is 29% to 73%. We report a case of 59-year-old female patient who was a known case of ALL, presented with the complaints of fever, lethargy and generalized weakness for one month. After taking blood samples for investigations, empirical antimicrobial therapy was started. She did not improve after 48 hours of therapy. Meanwhile blood culture revealed pure growth of A. hydrophila. After sensitivity report was available, ciprofloxacin was started. Patient became afebrile after 48 hours of treatment with ciprofloxacin. It is very vital to correctly identified and treat bacteremia due to A. hydrophila especially in the underlying leukemic patient. (author)

Classification of a hypervirulent Aeromonas hydrophila pathotype responsible for epidemic outbreaks in warm-water fishes

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Lineages of hypervirulent Aeromonas hydrophila (vAh) are the cause of persistent outbreaks of motile Aeromonas septicemia in warm-water fishes worldwide. Over the last decade, this virulent lineage of A. hydrophila has resulted in annual losses of millions of tons of farmed carp and catfish in the P...

Characterization of atypical Aeromonas salmonicida by different methods

DEFF Research Database (Denmark)

Austin, B.; Austin, D.A.; Dalsgaard, Inger

1998-01-01

Fifty two isolates of atypical Aeromonas salmonicida, recovered from a wide range of hosts and geographical locations, were heterogeneous in terms of molecular and phenotypic characteristics, and represented taxa which could not be accommodated by the current classification of four subspecies...

Antimicrobial resistance and plasmid profiles of Aeromonas ...

African Journals Online (AJOL)

The purpose of this study was to investigate the presence of Aeromonas hydrophila at commonly used water collection points on the River Njoro and to determine the in-vitro antimicrobial susceptibility and plasmid profiles of isolates. In total, 126 samples were collected and 36.5% of them were positive for A. hydrophila.

Pathogenic potentials of Aeromonas species isolated from aquaculture and abattoir environments.

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Igbinosa, Isoken H; Beshiru, Abeni; Odjadjare, Emmanuel E; Ateba, Collins N; Igbinosa, Etinosa O

2017-06-01

The present study elucidated the presence of antibiotics resistance, virulence genes and biofilm potentials among Aeromonas species isolated from abattoir and aquaculture environments in Benin City, Nigeria. A total of 144 wastewater samples were obtained from two independent aquaculture and abattoir environments between May and October 2016. Aeromonas species were isolated on Glutamate Starch Phenol Red (GSP) agar and confirmed using API 20NE kits. Antimicrobial susceptibility profile of the isolates was carried out using standard disc diffusion assay while biofilm potentials were detected by the microtitre plate method and PCR technique was used to detect antibiotics resistance and virulence gene markers. Overall, 32 and 26 Aeromonas species were isolated from the abattoir and aquaculture environments respectively. Isolates from both environments were completely resistant (100%) to penicillin G, ertapenem and tetracycline; whereas aquaculture isolates exhibited absolute sensitivity (100%) towards cefepime. All the virulence gene markers (aerA, hlyA, alt, ast, laf, ascF-G, fla, lip, stx1, and stx2) investigated in this study (except laf) were detected in isolates from both environments. The laf genes were only detected in isolates from abattoir environments. Antibiotics resistant genes including pse, bla TEM and class 1 integron were detected in isolates from both environments. Majority of the isolates (53/58 91.4%) from both environments; demonstrated capacity for biofilm potential. The detection of antibiotic resistance and virulence gene markers as well as biofilm forming ability in Aeromonas species isolated from aquaculture and abattoir environments raise serious public health concern worthy of further investigation. Copyright © 2017 Elsevier Ltd. All rights reserved.

Polymerase chain reaction (PCR)-based typing analysis of atypical isolates of the fish pathogen Aeromonas salmonicida

DEFF Research Database (Denmark)

Høie, S.; Dalsgaard, Inger; Aase, I.L.

1999-01-01

. salmonicida subsp. salmonicida isolates tested, belonged to group 1. The PCR primer-sets separated A. salmonicida from other reference strains of Aeromonas species and related bacteria with the exception of Aeromonas hydrophila. The results indicated that PCR typing is a useful framework for characterization...

Endoparasitos em cobaias (Cavia porcellus (Mammalia, Rodentia, Caviidae provenientes de biotérios de criação e experimentação do município do Rio de Janeiro, Brasil Endoparasites in guinea pigs (Cavia porcellus (Mammalia, Rodentia, Caviidae from breeding and experimentation animal housing of the municipality of Rio de Janeiro, Brazil

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Luciana Casartelli Alves

2007-10-01

Full Text Available Foi realizado um levantamento sobre a prevalência e intensidade de infecção de endoparasitos em cobaias convencionais de linhagem Short Hair provenientes de biotérios de criação (A e experimentação (B do município do Rio de Janeiro, Brasil, avaliando-se a eficácia das medidas de prevenção entre eles. Para a realização do estudo, utilizou-se exame direto de mucosa e do conteúdo intestinal, a técnica de tricromo de WHEATLEY e exames coproparasitológicos pelas técnicas de Ritchie e Kinyoun. Os parasitos encontrados através da técnica de exame direto da mucosa e do conteúdo intestinal e de tricromo de WHEATLEY com as respectivas prevalências foram: Balantidium sp. (78%, Cyathodinium sp. (68%, Eimeria caviae (38%, Paraspidodera uncinata (34% e Giardia muris (24%. Nos exames de fezes realizados pelas técnicas de Ritchie e Kinyoun, foram encontrados os seguintes parasitos com as respectivas prevalências no biotério A: E. caviae (74%, Balantidium sp. (68%, Cyathodinium sp. (68% e Cryptosporidium sp. (5%. No biotério B, observou-se: E. caviae (58%, Balantidium sp. (42%, Cyathodinium sp. (25% e G. muris (8%. A alta prevalência de endoparasitos nos biotérios sugere a necessidade de se rever a eficácia das barreiras sanitárias adotadas.This paper discusses the prevalence and intensity of infection of endoparasites in conventionally maintained Short Hair guinea pigs colonies from a breeding (A and an experimental (B facilities in Rio de Janeiro, Brazil. It also evaluates the efficacy of the methods of prevention adopted by both facilities. The search of parasites was performed by direct examination of intestinal mucosa and its contents, WHEATLEY's trichrome method and coproparasitological examinations by Ritchie's and Kinyoun's techniques. The prevalences of endoparasites found throught direct examination of intestinal mucosa and its contents were: Balantidium sp. (78%, Cyathodinium sp. (68%, Eimeria caviae (38%, Paraspidodera

Presence of Aeromonas spp in water from drinking-water- and wastewater-treatment plants in Mexico City.

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Villarruel-López, Angélica; Fernández-Rendón, Elizabeth; Mota-de-la-Garza, Lydia; Ortigoza-Ferado, Jorge

2005-01-01

The frequency of Aeromonas spp in three wastewater-treatment plants (WWTPs) and two drinking-water plants (DWPs) in México City was determined. Samples were taken throughout a year by the Moore's swab technique. A total of 144 samples were obtained from WWTPs and 96 from DWPs of both incoming and outflowing water. Aeromonas spp was isolated in 31% of the samples, from both kinds of sources. The technique used for the isolation of the pathogen was suitable for samples with high associate microbiota content and for those with a scarce microbial content. The presence of mesophilic-aerobic, coliform, and fecal-coliform organisms was investigated to determine whether there was any correlation with the presence of Aeromonas spp. Most samples from WWTP, which did not comply with the Mexican standards, had the pathogen, and some of the samples from the outflow of the DWP, which were within the limits set by the Mexican standards, also had Aeromonas spp. Most samples containing Aeromonas spp. had concentrations below 0.1 ppm residual chlorine, and the strains were resistant to 0.3 ppm, which supports the recommendation to increase the residual chlorine concentration to 0.5 to 1.0 ppm, as recommended by the Mexican standards.

Opposite cytokine synthesis by fibroblasts in contact co-culture with osteosarcoma cells compared with transwell co-cultures.

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David, Manu S; Kelly, Elizabeth; Zoellner, Hans

2013-04-01

We recently reported exchange of membrane and cytoplasm during contact co-culture between human Gingival Fibroblasts (h-GF) and SAOS-2 osteosarcoma cells, a process we termed 'cellular sipping' to reflect the manner in which cells become morphologically diverse through uptake of material from the opposing cell type, independent of genetic change. Cellular sipping is increased by Tumor Necrosis Factor-α (TNF-α), and we here show for the first time altered cytokine synthesis in contact co-culture supporting cellular sipping compared with co-culture where h-GF and SAOS-2 were separated in transwells. SAOS-2 had often undetectably low cytokine levels, while Interleukin-6 (IL-6), Granulocyte Colony Stimulating Factor (G-CSF) and Granulocyte-Macrophage Colony Stimulating Factor (GM-CSF) were secreted primarily by TNF-α stimulated h-GF and basic Fibroblast Growth Factor (FGF) was prominent in h-GF lysates (p cultures permitting cellular sipping had lower IL-6, G-CSF and GM-CSF levels, as well as higher lysate FGF levels compared with TNF-α treated h-GF alone (p cultures in transwells, with increased IL-6, G-CSF and GM-CSF levels (p cultures where cellular sipping occurs, potentially contributing to tumor inflammatory responses. Crown Copyright © 2013. Published by Elsevier Ltd. All rights reserved.

POTENSI TUMBUHAN BANGKAL (Nauclea Orientalis UNTUK PENGENDALIAN BAKTERI Aeromonas Hydrophila (POTENTIAL PLANT BANGKAL (Nauclea orientalis FOR CONTROL Aeromonas Hydrophila

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Siti Aisiah

2017-02-01

Full Text Available Tujuan penelitian ini adalah didapatkan metode pengendalian penyakit yang disebabkan oleh bakteri Aeromonas hydrophila pada budi daya ikan yang ramah lingkungan. Dalam penelitian ini dilakukan uji sensitivitas terhadap bakteri A. hydrophila, uji minimal konsentrasi menghambat  bakteri A. hydrophila (uji MIC dan uji toksisitas terhadap ikan nila. Rancangan  yang digunakan untuk  uji toksisitas  adalah rancangan acak lengkap (RAL, dengan 4 perlakuan yaitu A  = Ikan disuntik dengan ekstrak bangkal konsentrasi 20%, B  = Ikan disuntik dengan ekstrak bangkal konsentrasi 40%,  C  = Ikan disuntik dengan ekstrak bangkal konsentrasi 80%, dan D  = Kontrol (ikan tidak disuntik, diulang sebanyak 3 kali. Perlakuan ini didapat dari hasil uji sensitivitas antibakteri bangkal yang mempunyai daya hambat dan daya bunuh paling besar terhadap bakteri A. hydrophila yaitu ekstrak daun bangkal dengan pelarut akuades. Pengujian MIC menunjukkan bahwa ekstrak bangkal-akuades memiliki daya hambat minimal  20 % terhadap aktivitas bakteri A. hydrophila. Hasil uji toksisitas yang dilakukan terhadap ikan nila dengan konsentrasi 20%, 40% dan 80%  mortalitas ikan nila dibawah 50 %. Pengamatan hematologis yaitu eretrosit, leokosit, plasma darah, hematokrit dan leokokrit pada masing-masing perlakuan sebagian besar masih berada dalam kisaran yang normal. Parameter kualitas air yaitu, kadar oksigen terlarut,  pH, amoniak, CO2 dan suhu masih dapat mendukung kehidupan normal ikan nila. The purpose of this study was obtained method of controlling diseases caused by Aeromonas hydrophila in fish farming is environmentally friendly. In this study the sensitivity of the test  A. hydrophila, a minimum test konsentari inhibiting A. hydrophila (MIC test and toxicity test on  tilapia. The design used for toxicity tests is completely randomized design with 4 treatments it A = Fish injected with extracts bangkal concentration of 20 %,  B = Fish injected with extracts bangkal

Evidence of Increased Antibiotic Resistance in Phylogenetically-Diverse Aeromonas Isolates from Semi-Intensive Fish Ponds Treated with Antibiotics.

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Patil, Hemant J; Benet-Perelberg, Ayana; Naor, Alon; Smirnov, Margarita; Ofek, Tamir; Nasser, Ahmed; Minz, Dror; Cytryn, Eddie

2016-01-01

The genus Aeromonas is ubiquitous in aquatic environments encompassing a broad range of fish and human pathogens. Aeromonas strains are known for their enhanced capacity to acquire and exchange antibiotic resistance genes and therefore, are frequently targeted as indicator bacteria for monitoring antimicrobial resistance in aquatic environments. This study evaluated temporal trends in Aeromonas diversity and antibiotic resistance in two adjacent semi-intensive aquaculture facilities to ascertain the effects of antibiotic treatment on antimicrobial resistance. In the first facility, sulfadiazine-trimethoprim was added prophylactically to fingerling stocks and water column-associated Aeromonas were monitored periodically over an 11-month fish fattening cycle to assess temporal dynamics in taxonomy and antibiotic resistance. In the second facility, Aeromonas were isolated from fish skin ulcers sampled over a 3-year period and from pond water samples to assess associations between pathogenic strains to those in the water column. A total of 1200 Aeromonas isolates were initially screened for sulfadiazine resistance and further screened against five additional antimicrobials. In both facilities, strong correlations were observed between sulfadiazine resistance and trimethoprim and tetracycline resistances, whereas correlations between sulfadiazine resistance and ceftriaxone, gentamicin, and chloramphenicol resistances were low. Multidrug resistant strains as well as sul1, tetA , and intI1 gene-harboring strains were significantly higher in profiles sampled during the fish cycle than those isolated prior to stocking and these genes were extremely abundant in the pathogenic strains. Five phylogenetically distinct Aeromonas clusters were identified using partial rpoD gene sequence analysis. Interestingly, prior to fingerling stocking the diversity of water column strains was high, and representatives from all five clusters were identified, including an A. salmonicida

Differences in detection of Aeromonas salmonicida in covertly infected salmonid fishes by the stress-inducible furunculosis test and culture-based assays

Science.gov (United States)

Cipriano, R.C.; Ford, L.A.; Smith, D.R.; Schachte, J.H.; Petrie, C.J.

1997-01-01

Accurate detection of Aeromonas salmonicida subsp. salmonicida (the cause of furunculosis disease) in covertly infected salmonids is difficult and is a cause of concern for those involved in fish health inspection and resource management programs. In this study, we examined populations of brook trout Salvelinus fontinalis, Atlantic salmon Salmo salar, and lake trout Salvelinus namaycush that previously sustained natural episodes of furunculosis. Consequently, the sampled fish were presumed to harbor latent infections. Mucus, gill, liver, kidney, heart, spleen, and intestine samples (N = 100 fish per group sampled) were processed and examined by (1) direct dilution counts and (2) quadrant streaking after a 48-h pre-enrichment in trypticase soy broth (TSB). Another subsample of fish from each group was then subjected to stress-inducible furunculosis tests. Stress tests detected A. salmonicida in three of four groups of fish that were examined whereas the pathogen was detected in only two of the groups analyzed with culture-based assays. Although pre-enrichment in TSB enhanced detection within internal sampling sites including the liver, heart, spleen, and kidney, enrichment did not enhance detection from mucus, gill, or intestinal samples.

Reciprocal immune benefit based on complementary production of antibiotics by the leech Hirudo verbana and its gut symbiont Aeromonas veronii.

Science.gov (United States)

Tasiemski, Aurélie; Massol, François; Cuvillier-Hot, Virginie; Boidin-Wichlacz, Céline; Roger, Emmanuel; Rodet, Franck; Fournier, Isabelle; Thomas, Frédéric; Salzet, Michel

2015-12-04

The medicinal leech has established a long-term mutualistic association with Aeromonas veronii, a versatile bacterium which can also display free-living waterborne and fish- or human-pathogenic lifestyles. Here, we investigated the role of antibiotics in the dynamics of interaction between the leech and its gut symbiont Aeromonas. By combining biochemical and molecular approaches, we isolated and identified for the first time the antimicrobial peptides (AMPs) produced by the leech digestive tract and by its symbiont Aeromonas. Immunohistochemistry data and PCR analyses evidenced that leech AMP genes are induced in the gut epithelial cells when Aeromonas load is low (starved animals), while repressed when Aeromonas abundance is the highest (post blood feeding). The asynchronous production of AMPs by both partners suggests that these antibiotic substances (i) provide them with reciprocal protection against invasive bacteria and (ii) contribute to the unusual simplicity of the gut microflora of the leech. This immune benefit substantially reinforces the evidence of an evolutionarily stable association between H. verbana and A. veronii. Altogether these data may provide insights into the processes making the association with an Aeromonas species in the digestive tract either deleterious or beneficial.

Aeromonas hydrophila disturbs water and electrolyte transport in ...

African Journals Online (AJOL)

SERVER

2008-02-19

Feb 19, 2008 ... enterocytes with voluminous and spherical shape, disappearance of enterocyte brush border and lesions at ... (Paniagua et al., 1990) and causes different diseases. ... 22°C and the fish were fed on a standard diet composed of algae. ..... J (1998). Aeromonas hydrophila causes 'black diseases' in fairy.

Population dynamics, antibiotics resistance and biofilm formation of Aeromonas and Vibrio species isolated from aquatic sources in Northern Malaysia.

Science.gov (United States)

Odeyemi, Olumide A; Ahmad, Asmat

2017-02-01

This study aimed to compare population dynamics, antibiotic resistance and biofilm formation of Aeromonas and Vibrio species from seawater and sediment collected from Northern Malaysia. Isolates with different colony morphology were characterized using both biochemical and molecular methods before testing for antibiotic resistance and biofilm formation. Results obtained from this study showed that in Kedah, the population of Aeromonas isolated from sediment was highest in Pantai Merdeka (8.22 log CFU/ml), Pulau Bunting recorded the highest population of Aeromonas from sediment (8.43 log CFU/g). It was observed that Vibrio species isolated from seawater and sediment were highest in Kuala Sanglang (9.21 log CFU/ml). In Kuala Perlis, the population of Aeromonas isolated from seawater was highest in Jeti (7.94 log CFU/ml). Highest population of Aeromonas from sediment was recorded in Kampong Tanah Baru (7.99 log CFU/g). It was observed that Vibrio species isolated from seawater was highest in Padang Benta (8.42 log CFU/g) while Jeti Kuala Perlis had highest population of Vibrio isolated from sediment. It was observed that location does not influence population of Aeromonas. The results of the independent t - test revealed that there was no significant relationship between location and population of Vibrio (df = 10, t = 1.144, p > 0.05). The occurrence of biofilm formation and prevalence of antibiotic resistant Aeromonas and Vibrio species in seawater and sediment pose danger to human and aquatic animals' health. Copyright © 2017 Elsevier Ltd. All rights reserved.

CONTAMINATION RISK ASSESSMENT OF SOURCES OF WATER INTENDED FOR HUMAN CONSUMPTION WITH AEROMONAS SPP.

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Luminita Minea

2012-03-01

Full Text Available mportance of Aeromonas spp as emerging pathogens in recent years has been a considerable increase, beingincluded by the World Health Organization (WHO and Environmental Protection Agency (EPA in the etiology ofwaterborne epidemics. We aimed to investigate the distribution of Aeromonas spp. in various aquatic ecosystems whichcan become potential sources of gastrointestinal infections. Simultaneously, the presence of this pathogen was evaluatedin relation to microbiological indicators set out in EU Directive 98/83/EC. Between March 2009 and May 2011 wereanalyzed by membrane filter technique untreated water samples, collected from surface sources (lakes, ponds andgroundwater (wells drilled, captured springs, wells and chlorinated water that coming from local distribution network,located in rural areas of Moldova (North-East Romania. The information was processed and interpreted statisticallyusing EpiInfo 3.5.1 software. The presence of Aeromonas spp. was detected in half of the 220 samples tested, especially,in surface waters (84% and captured springs (80.8% (χ 2=51.39; GL=4; p0.8, being influenced by exogenous humanimpact. The results of this study confirms that untreated water used in rural areas can affect public health andmicrobiological indicators have predictive value for the presence of Aeromonas spp only reporting to surface sources.

Thin-film fixed-bed reactor for solar photocatalytic inactivation of Aeromonas hydrophila: influence of water quality

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Khan Sadia J

2012-11-01

Full Text Available Abstract Background Controlling fish disease is one of the major concerns in contemporary aquaculture. The use of antibiotics or chemical disinfection cannot provide a healthy aquaculture system without residual effects. Water quality is also important in determining the success or failure of fish production. Several solar photocatalytic reactors have been used to treat drinking water or waste water without leaving chemical residues. This study has investigated the impact of several key aspects of water quality on the inactivation of the pathogenic bacterium Aeromonas hydrophila using a pilot-scale thin-film fixed-bed reactor (TFFBR system. Results The level of inactivation of Aeromonas hydrophila ATCC 35654 was determined using a TFFBR with a photocatalytic area of 0.47 m2 under the influence of various water quality variables (pH, conductivity, turbidity and colour under high solar irradiance conditions (980–1100 W m-2, at a flow rate of 4.8 L h-1 through the reactor. Bacterial enumeration were obtained through conventional plate count using trypticase soy agar media, cultured in conventional aerobic conditions to detect healthy cells and under ROS-neutralised conditions to detect both healthy and sub-lethally injured (oxygen-sensitive cells. The results showed that turbidity has a major influence on solar photocatalytic inactivation of A. hydrophila. Humic acids appear to decrease TiO2 effectiveness under full sunlight and reduce microbial inactivation. pH in the range 7–9 and salinity both have no major effect on the extent of photoinactivation or sub-lethal injury. Conclusions This study demonstrates the effectiveness of the TFFBR in the inactivation of Aeromonas hydrophila under the influence of several water quality variables at high solar irradiance, providing an opportunity for the application of solar photocatalysis in aquaculture systems, as long as turbidity remains low.

Paraspidodera uncinata (Nematoda, Lauroiinae as parasite of Cavia magna and Cavia aperea (Rodentia, Caviidae in southern Brazil

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Ana Gabriela da Silva Rocha

2015-05-01

Full Text Available Out of the 5 species of Guinea pig registered for Brazil, 4 (Cavia fulgida, C. porcellus, C. aperea, and C. magna have already been reported as hosts of Paraspidodera uncinata. The rodent species C. magna and C. aperea are small-sized mammals, with terrestrial habits, which occur from southern Brazil to eastern Uruguay. Guinea pig specimens donated for research were necropsied for the analysis of endoparasites, and the nematodes found were identified as P. uncinata. The microhabitats of this species were the small and large intestines in C. magna and the large intestine in C. aperea. The prevalence of P. uncinata in the hosts was 60% in C. magna (n = 5 and 14% in C. aperea (n = 7. The record of P. uncinata in different Guinea pig species allows inferring that the foraging mode is similar in the different locations where they occur in southern Brazil, because, perhaps, the infection of hosts has occurred through the ingestion of vegetables contaminated with P. uncinata eggs. This study contributes to knowledge on the helminth fauna of C. magna and C. aperea in southern Brazil.

Simultaneous effects of environmental factors on motile Aeromonas dynamics in an urban effluent and in the natural seawater.

Science.gov (United States)

Maalej, Sami; Mahjoubi, Amira; Elazri, Chafai; Dukan, Sam

2003-07-01

Seasonal dynamics of motile Aeromonas in a treated urban effluent and in natural seawater along the Sfax coast (Mediterranean sea, Tunisia) were measured over a year concurrently with seven environmental factors, and compared with those of faecal coliforms. Counts for Aeromonas from a standard plate count method, ranged from 1.48 x 10(5)CFU.100 ml(-1) to 2.2 x 10(8)CFU.100 ml(-1) in the effluent and from 7.9 x 10(3)CFU.100 ml(-1) to undetectable level in the surface marine waters. Contrary to faecal coliforms, the Aeromonas dynamics exhibited a seasonal distribution in seawater which was inverse of the seasonal distribution in the sewage: From the end of November 1998 to April 1999 (cold period), Aeromonas counts increased in the treated effluent, while it decreased very rapidly in seawater. From May to October (warm period), Aeromonas abundance decreased in the effluent but showed an increasing fluctuating trend in the marine waters with a maximum in late summer/early autumn when the temperatures were around 22-23 degrees C. Multiple correlation and regression analyses suggest, by the coefficient of determination (R(2)), that 42% of variance in Aeromonas number changes in the treated effluent, may be explained by only turbidity, radiation and Aeromonas density in the previous sample, while 37% of variance in marine ecosystem were explained by radiance and conductivity. Furthermore, the t statistics and their p values and the coefficient of partial determination (r(2)) indicated that radiance contributed the most (r(2)=0.3184, t=-3.2, p=0.0041) to the dynamics of motile Aeromonas in seawater, when combined with conductivity. The models relevant for changes in faecal coliforms abundance incorporated turbidity, radiance in the effluent and conductivity, pH, radiance, turbidity in coastal marine environment. These models explain 66% and 73% of the observed cell number fluctuation, with turbidity (r(2)=0.529, t=5.08, p=0.0001) and conductivity (r(2)=0.5407, t=4.97, p=0

Aeromonas hydrophila disturbs water and electrolyte transport in ...

African Journals Online (AJOL)

Fish diseases create a menace to aquaculture farms. They provoke disastrous economic losses and sanitary risks for the consumer. The present study aims to investigate the effect of the bacteria, Aeromonas hydrophila on water and electrolyte (Na+, K+, Cl-, HCO3 -) flux of Mugil cephalus (L, 1758) intestine. Anterior, middle ...

Meningoencephalitis in farmed monosex Nile tilapia (Oreochromis niloticus L. caused by Streptococcus agalactiae

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Adikesavalu Harresh

2017-09-01

Full Text Available Aquaculture of tilapia is a new research venture in India. With intensification in farming practices, tilapia are increasingly susceptible to bacterial infections. This article describes the isolation and identification of pathogenic bacteria from cultured monosex Nile tilapia, Oreochromis niloticus (L., that experienced moderate to severe mortalities in West Bengal, India between September and August 2014 and histopathological alterations in various organs. Gram-positive diplococci, identified as Streptococcus agalactiae with Streptococcus identification kits and 16S rDNA sequencing analysis, were isolated from the brain, operculum, and kidney. Other bacteria from the kidney were identified as Aeromonas sobria, A. caviae, Klebsiella pneumoniae ssp. pneumoniae, Escherichia coli, and Enterobacter cloacae. Staphylococcus epidermis was isolated from opercular hemorrhages. Histological sections of the infected tilapia brain revealed meningoencephalitis and granulomatous lesions. Sections from other organs indicated congestion, hemorrhagic and hyperplastic cells, necrosis, vacuolation, hemosiderin deposition, hypertrophic nuclei, melanomacrophage aggregation, and ruptured veins. This report is the first description of S. agalactiae as a primary pathogen causing meningoencephalitis in cultured tilapia in India.

Helminths of the guinea pig, Cavia porcellus (Linnaeus, in Brazil

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Roberto Magalhães Pinto

2002-07-01

Full Text Available Worm burdens were evaluated and compared in two groups of the guinea pig, Cavia poreellus (Linnaeus, 1758: animals of the first group were conventionally maintained in an institutional animal house and those of the second group were openly kept in pet shops in Brazil. Animals from both sources were infected only with the nematode Paraspidodera uncinata (Rudolphi, 1819 Travassos, 1914 (10% of prevalence in guinea pigs from lhe institutional facility and 40% in those animals from the pet shop. Other helminth samples recovered from Brazilian guinea pigs during 52 years and that are deposited in the Helminthological Collection of the Oswaldo Cruz Institute (CHIOC were also analyzed. Paraspidodera uncinata and the cestode Monoecocestus parcitesticulatus Rêgo, 1960 were identified in these samples.

Jejuno-jejunal intussusception in a guinea pig (Cavia porcellus

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Tara J. Fetzer

2017-09-01

Full Text Available An approximately four-year-old male castrated guinea pig (Cavia porcellus was presented for painful defecation with a 24-hour history of hyporexia and intermittent episodes of rolling behavior. Upon presentation the patient was quiet, alert, and responsive, and mildly hypothermic. Abdominal palpation revealed an approximately 2-cm long oblong mass within the caudal abdomen. Abdominal radiographs revealed gastric dilation without volvulus and a peritoneal mass effect. The patient was euthanized following gastric reflux of brown malodorous fluid from his nares and oral cavity. A necropsy was performed and revealed a jejuno-jejunal intussusception causing mechanical gastrointestinal ileus, and gastric dilatation without volvulus. While non-obstructive gastrointestinal stasis is common and obstructive ileus is uncommon in guinea pigs, this report shows that intestinal intussusception is a differential in guinea pigs with ileus and gastric dilatation.

The Effect of Some Physical and Chemical Parameters on Regrowth of Aeromonas Bacterium and Heterotrophic Bacteria in Isfahan Drinking Water System

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Sina Dobaradaran

2006-03-01

Full Text Available Aeromonas is one the gram – negative , non spor – formating rod shaping , facultatively anaerobic and opportunistic bactria that can cause systematic infections, leision and diarrhoea in human. Fairly high bactrial population in distribution system is not only of concern because of affecting consumer health but also it makes it difficult to enumerate coliform bacterium indicator. So, the relationships between aeromonas and heterotrophic bacteria growth with pH, temperature, turbidity, free residual cholornie and DO were determined in this study. ADA- V media was used in presumptive stage to count aeromonas bacteria for the first time in Iran on the basis of 1605 EPA (2001 method and used oxidase tests, trehalose fermentation and indol test in confirmative stage. R2A media was used to count HPC bacteria and other factors measured on the basis of standards. The results showed that positive cases of aeromonas bacteria and HPC increase in higher temperature and turbidity and lower pH. In contrast, positive cases of aeromonas bacteria and HPC decrease while free residual cholorine and DO increase. In addition,no positive case of aeromonas was observed in more than 0.2 mg/L concentration of free residual cholorine.

Evidence of increased antibiotic resistance in phylogenetically-diverse Aeromonas isolates from semi-intensive fish ponds treated with antibiotics

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Hemant J Patil

2016-11-01

Full Text Available The genus Aeromonas is ubiquitous in aquatic environments encompassing a broad range of fish and human pathogens. Aeromonas strains are known for their enhanced capacity to acquire and exchange antibiotic resistance genes and therefore, are frequently targeted as indicator bacteria for monitoring antimicrobial resistance in aquatic environments. This study evaluated temporal trends in Aeromonas diversity and antibiotic resistance in two adjacent semi-intensive aquaculture facilities to ascertain effects of antibiotic treatment on antimicrobial resistance. In the first facility, sulfadiazine-trimethoprim was added prophylactically upon fingerling stocking and water column-associated Aeromonas were monitored periodically over an eleven-month fish-fattening cycle to assess temporal dynamics in taxonomy and antibiotic resistance. In the second facility, Aeromonas were isolated from fish skin ulcers sampled over a three-year period and from pond water samples to assess associations between pathogenic strains to those in the water column. A total of 1200 Aeromonas spp. were isolated, initially screened for sulfadiazine resistance and further screened against five additional antibiotics. In both facilities, strong correlations were observed between sulfadiazine resistance and trimethoprim and tetracycline resistances, whereas correlations between sulfadiazine resistance and ceftriaxone, gentamycin and chloramphenicol resistances were low. Abundance of multi-drug resistant strains as well as sul1, tetA and intI1 gene-harboring strains was significantly higher in profiles sampled during the fish cycle than those isolated prior to stocking and these genes were extremely abundant in the pathogenic strains. Five phylogenetically-distinct Aeromonas clusters were revealed using partial rpoD gene sequence analysis. Interestingly, prior to fingerling stocking the diversity of water column strains was high, and representatives from all five clusters were

Insight into the mobilome of Aeromonas strains.

Science.gov (United States)

Piotrowska, Marta; Popowska, Magdalena

2015-01-01

The mobilome is a pool of genes located within mobile genetic elements (MGE), such as plasmids, IS elements, transposons, genomic/pathogenicity islands, and integron-associated gene cassettes. These genes are often referred to as "flexible" and may encode virulence factors, toxic compounds as well as resistance to antibiotics. The phenomenon of MGE transfer between bacteria, known as horizontal gene transfer (HGT), is well documented. The genes present on MGE are subject to continuous processes of evolution and environmental changes, largely induced or significantly accelerated by man. For bacteria, the only chance of survival in an environment contaminated with toxic chemicals, heavy metals and antibiotics is the acquisition of genes providing the ability to survive in such conditions. The process of acquiring and spreading antibiotic resistance genes (ARG) is of particular significance, as it is important for the health of humans and animals. Therefore, it is important to thoroughly study the mobilome of Aeromonas spp. that is widely distributed in various environments, causing many diseases in fishes and humans. This review discusses the recently published information on MGE prevalent in Aeromonas spp. with special emphasis on plasmids belonging to different incompatibility groups, i.e., IncA/C, IncU, IncQ, IncF, IncI, and ColE-type. The vast majority of plasmids carry a number of different transposons (Tn3, Tn21, Tn1213, Tn1721, Tn4401), the 1st, 2nd, or 3rd class of integrons, IS elements (e.g., IS26, ISPa12, ISPa13, ISKpn8, ISKpn6) and encode determinants such as antibiotic and mercury resistance genes, as well as virulence factors. Although the actual role of Aeromonas spp. as a human pathogen remains controversial, species of this genus may pose a serious risk to human health. This is due to the considerable potential of their mobilome, particularly in terms of antibiotic resistance and the possibility of the horizontal transfer of resistance genes.

Insight into the mobilome of Aeromonas strains

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Marta ePiotrowska

2015-05-01

Full Text Available The mobilome is a pool of genes located within mobile genetic elements (MGE, such as plasmids, IS elements, transposons, genomic/pathogenicity islands, and integron-associated gene cassettes. These genes are often referred to as flexible and may encode virulence factors, toxic compounds as well as resistance to antibiotics. The phenomenon of MGE transfer between bacteria, known as horizontal gene transfer (HGT, is well documented. The genes present on MGE are subject to continuous processes of evolution and environmental changes, largely induced or significantly accelerated by man. For bacteria, the only chance of survival in an environment contaminated with toxic chemicals, heavy metals and antibiotics is the acquisition of genes providing the ability to survive in such conditions. The process of acquiring and spreading antibiotic resistance genes (ARG is of particular significance, as it is important for the health of humans and animals. Therefore, it is important to thoroughly study the mobilome of Aeromonas spp. that is widely distributed in various environments, causing many diseases in fishes and humans. This review discusses the recently published information on MGE prevalent in Aeromonas spp. with special emphasis on plasmids belonging to different incompatibility groups, i.e., IncA/C, IncU, IncQ, IncF, IncI and ColE-type. The vast majority of plasmids carry a number of different transposons (Tn3, Tn21, Tn1213, Tn1721, Tn4401, the 1st, 2nd or 3rd class of integrons, IS elements (e.g. IS26, ISPa12, ISPa13, ISKpn8, ISKpn6 and encode determinants such as antibiotic and mercury resistance genes, as well as virulence factors. Although the actual role of Aeromonas spp. as a human pathogen remains controversial, species of this genus may pose a serious risk to human health. This is due to the considerable potential of their mobilome, particularly in terms of antibiotic resistance and the possibility of the horizontal transfer of resistance

Aeromonas spp. in het Nederlandse drinkwater; een orienterend onderzoek

NARCIS (Netherlands)

Versteegh JFM; During M; Havelaar AH; Koot W

1986-01-01

In dit rapport worden uitvoering en resultaten van een orienterend onderzoek naar het voorkomen van Aeromonas spp. in het Nederlandse drinkwater in 1985 besproken. Het bleek dat in 71% van de onderzochte monsters (rein water "af pompstation" en water uit het distributienet) in 100 ml

Ciprofloxacin-Resistant Aeromonas hydrophila Cellulitis following Leech Therapy

Science.gov (United States)

Giltner, Carmen L.; Bobenchik, April M.; Uslan, Daniel Z.; Deville, Jaime G.

2013-01-01

We report a case of surgical site infection with ciprofloxacin-resistant Aeromonas hydrophila following leech therapy. Antimicrobial and genetic analyses of leech and patient isolates demonstrated that the resistant isolates originated from the leech gut microbiota. These data suggest that ciprofloxacin monotherapy as a prophylaxis regimen prior to leech therapy may not be effective in preventing infection. PMID:23363826

Trixacarus caviae Fain, Howell & Hyatt 1972 (Acari: Sarcoptidae) as a cause of mange in guinea-pigs and papular urticaria in man

NARCIS (Netherlands)

Dorrestein, G.M.; Bronswijk, J.E.M.H. van

1979-01-01

The clinical symptoms of infestation with Trixacarus caviae in guinea-pigs are described. In general large areas of thickened, denuded skin, with a heavy secondary bacterial infection, and sometimes neurological signs, are observed in mangy cases. Antiparasitic therapy was successful with one or

Application of a continuously stirred tank bioreactor (CSTR) for bioremediation of hydrocarbon-rich industrial wastewater effluents

International Nuclear Information System (INIS)

Gargouri, Boutheina; Karray, Fatma; Mhiri, Najla; Aloui, Fathi; Sayadi, Sami

2011-01-01

A continuously stirred tank bioreactor (CSTR) was used to optimize feasible and reliable bioprocess system in order to treat hydrocarbon-rich industrial wastewaters. A successful bioremediation was developed by an efficient acclimatized microbial consortium. After an experimental period of 225 days, the process was shown to be highly efficient in decontaminating the wastewater. The performance of the bioaugmented reactor was demonstrated by the reduction of COD rates up to 95%. The residual total petroleum hydrocarbon (TPH) decreased from 320 mg TPH l -1 to 8 mg TPH l -1 . Analysis using gas chromatography-mass spectrometry (GC-MS) identified 26 hydrocarbons. The use of the mixed cultures demonstrated high degradation performance for hydrocarbons range n-alkanes (C10-C35). Six microbial isolates from the CSTR were characterized and species identification was confirmed by sequencing the 16S rRNA genes. The partial 16S rRNA gene sequences demonstrated that 5 strains were closely related to Aeromonas punctata (Aeromonas caviae), Bacillus cereus, Ochrobactrum intermedium, Stenotrophomonas maltophilia and Rhodococcus sp. The 6th isolate was affiliated to genera Achromobacter. Besides, the treated wastewater could be considered as non toxic according to the phytotoxicity test since the germination index of Lepidium sativum ranged between 57 and 95%. The treatment provided satisfactory results and presents a feasible technology for the treatment of hydrocarbon-rich wastewater from petrochemical industries and petroleum refineries.

Application of a continuously stirred tank bioreactor (CSTR) for bioremediation of hydrocarbon-rich industrial wastewater effluents

Energy Technology Data Exchange (ETDEWEB)

Gargouri, Boutheina; Karray, Fatma; Mhiri, Najla; Aloui, Fathi [Laboratoire des Bioprocedes Environnementaux, Pole d' Excellence Regional AUF-LBPE, Centre de Biotechnologie de Sfax, Universite de Sfax, BP 1117, 3018 Sfax (Tunisia); Sayadi, Sami, E-mail: sami.sayadi@cbs.rnrt.tn [Laboratoire des Bioprocedes Environnementaux, Pole d' Excellence Regional AUF-LBPE, Centre de Biotechnologie de Sfax, Universite de Sfax, BP 1117, 3018 Sfax (Tunisia)

2011-05-15

A continuously stirred tank bioreactor (CSTR) was used to optimize feasible and reliable bioprocess system in order to treat hydrocarbon-rich industrial wastewaters. A successful bioremediation was developed by an efficient acclimatized microbial consortium. After an experimental period of 225 days, the process was shown to be highly efficient in decontaminating the wastewater. The performance of the bioaugmented reactor was demonstrated by the reduction of COD rates up to 95%. The residual total petroleum hydrocarbon (TPH) decreased from 320 mg TPH l{sup -1} to 8 mg TPH l{sup -1}. Analysis using gas chromatography-mass spectrometry (GC-MS) identified 26 hydrocarbons. The use of the mixed cultures demonstrated high degradation performance for hydrocarbons range n-alkanes (C10-C35). Six microbial isolates from the CSTR were characterized and species identification was confirmed by sequencing the 16S rRNA genes. The partial 16S rRNA gene sequences demonstrated that 5 strains were closely related to Aeromonas punctata (Aeromonas caviae), Bacillus cereus, Ochrobactrum intermedium, Stenotrophomonas maltophilia and Rhodococcus sp. The 6th isolate was affiliated to genera Achromobacter. Besides, the treated wastewater could be considered as non toxic according to the phytotoxicity test since the germination index of Lepidium sativum ranged between 57 and 95%. The treatment provided satisfactory results and presents a feasible technology for the treatment of hydrocarbon-rich wastewater from petrochemical industries and petroleum refineries.

Avaliação da qualidade microbiológica de águas minerais consumidas na região metropolitana de Recife, Estado de Pernambuco = Evaluation of the microbiological quality of mineral water consumed in the metropolitan region of Recife, Pernambuco State

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Marcelo Iran Souza Coelho

2010-01-01

Full Text Available Águas minerais comercializadas em garrafões de 20 L na região metropolitana de Recife, Estado de Pernambuco foram analisadas microbiologicamente, no período de junho de 2002 a maio de 2003. As dez marcas escolhidas de água mineral foram avaliadas no tocante às bactérias heterotróficas, coliformes, Escherichia coli, Pseudomonas spp., Pseudomonas aeruginosa e Aeromonas spp. Os dados foram analisados estatisticamente por meio de Modelos Lineares Generalizados (GLM. As águas minerais apresentaram contaminação por E. coli, P. aeruginosa, A. hydrophila e A. caviae. Concluiu-se que, de todas as dez marcas analisadas, pelo menos uma amostra de cada marca apresentou-se imprópria para o consumo, por não atender aos parâmetros estabelecidos pela legislação em vigor em, no mínimo, duas análises; o Número Mais Provável (NMP de Pseudomonas spp. e de P. aeruginosa foi influenciado pela sazonalidade e a contagem de bactérias heterotróficas; o Número Mais Provável (NMP de coliformes totais e termotolerantes e a pesquisa de Aeromonas spp. não sofreram influência significativa da sazonalidade.Mineral water sold in 20 L bottles in the metropolitan area of Recife, Pernambuco State were microbiologically analyzed between June 2002 and May 2003. The ten selected brands of mineral water were evaluated with regard to heterotrophic bacteria, coliforms, Escherichia coli, Pseudomonas spp., Pseudomonas aeruginosa and Aeromonas spp. Data were statistically analyzed using Generalized Linear Models (GLM. Mineral waters showed contamination by the following: E. coli, P. aeruginosa, A. hydrophila and A. caviae. It was concluded that of all ten tested brands, at least one sample of each brand was improper for consumption, for not meeting the parameters established by legislation in at least two tests; the Most Probable Number (MPN for Pseudomonas spp. and P. aeruginosa was influenced by seasonality and the count of heterotrophic bacteria; the Most

A co-cultured skin model based on cell support membranes

International Nuclear Information System (INIS)

Dai, N.-T.; Yeh, M.-K.; Liu, Demeral David; Adams, E.F.; Chiang, C.-H.; Yen, C.-Y.; Shih, C.-M.; Sytwu, H.-K.; Chen, Tim-Mo; Wang, H.-J.; Williamson, M.R.; Coombes, A.G.A.

2005-01-01

Tissue engineering of skin based on collagen: PCL biocomposites using a designed co-culture system is reported. The collagen: PCL biocomposites having collagen: PCL (w/w) ratios of 1:4, 1:8, and 1:20 have been proven to be biocompatible materials to support both adult normal human epidermal Keratinocyte (NHEK) and mouse 3T3 fibroblast growth in cell culture, respectively, by Dai, Coombes, et al. in 2004. Films of collagen: PCL biocomposites were prepared using non-crosslinking method by impregnation of lyophilized collagen mats with PCL/dichloromethane solutions followed by solvent evaporation. To mimic the dermal/epidermal structure of skin, the 1:20 collagen: PCL biocomposites were selected for a feasibility study of a designed co-culture technique that would subsequently be used for preparing fibroblast/biocomposite/keratinocyte skin models. A 55.3% increase in cell number was measured in the designed co-culture system when fibroblasts were seeded on both sides of a biocomposite film compared with cell culture on one surface of the biocomposite in the feasibility study. The co-culture of human keratinocytes and 3T3 fibroblasts on each side of the membrane was therefore studied using the same co-culture system by growing keratinocytes on the top surface of membrane for 3 days and 3T3 fibroblasts underneath the membrane for 6 days. Scanning electron microscopy (SEM) and immunohistochemistry assay revealed good cell attachment and proliferation of both human keratinocytes and 3T3 fibroblasts with these two types of cells isolated well on each side of the membrane. Using a modified co-culture technique, a co-cultured skin model presenting a confluent epidermal sheet on one side of the biocomposite film and fibroblasts populated on the other side of the film was developed successfully in co-culture system for 28 days under investigations by SEM and immunohistochemistry assay. Thus, the design of a co-culture system based on 1:20 (w/w) collagen: PCL biocomposite

Mass culture strategy for bacterial yeast co-culture for degradation of petroleum hydrocarbons in marine environment.

Science.gov (United States)

Priya, Anchal; Mandal, Ajoy K; Ball, Andrew S; Manefield, Mike; Lal, Banwari; Sarma, Priyangshu M

2015-11-15

In the present study a metabolically versatile co-culture with two Bacilli and one yeast strain was developed using enrichment culture techniques. The developed co-culture had affinity to degrade both aliphatic and aromatic fractions of petroleum crude oil. Degradation kinetics was established for designing the fermentation protocol of the co-culture. The developed mass culture strategy led to achieve the reduction in surface tension (26dynescm(-1) from 69 dynescm(-1)) and degradation of 67% in bench scale experiments. The total crude oil degradation of 96% was achieved in 4000l of natural seawater after 28days without adding any nutrients. The survival of the augmented co-culture was maintained (10(9)cellsml(-1)) in contaminated marine environment. The mass culture protocol devised for the bioaugmentation was a key breakthrough that was subsequently used for pilot scale studies with 100l and 4000l of natural seawater for potential application in marine oil spills. Copyright © 2015 Elsevier Ltd. All rights reserved.

The occurrence of Aeromonas spp. in the bottled mineral water, well water and tap water from the municipal supplies

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Denise de Oliveira Scoaris

2008-10-01

Full Text Available The aim of this work was to study the occurrence of Aeromonas sp in the bottled mineral water, well water and tap water from the municipal supplies. Positive samples were found for Aeromonas spp. 12.7% from the mineral water, 8.3% from the artesian water and 6.5% from the tap water. The recovery of Aeromonas spp. was significantly higher in the bottled mineral and artesian water than in the tap water from municipal supplies. The occurrence of the Aeromonas spp. did not correlate significantly with the contamination indicator bacteria (i.e. total coliforms in the artesian water samples. However, a significant correlation was found between Aeromonas spp. and total coliforms in the both mineral water and tap water samples. The presence or absence of a correlation between the indicator bacteria and Aeromonas could reflect the occasional appearance of the pathogen in the drinking water and the different rates of survival and recovery of these agents compared with those fecal indicators. The finding that 41.6, 14.8 and 9.0 % of the artesian water, bottled mineral water and tap water, respectively, sampled in the current study failed to meet the Brazilian standard for total coliforms in the drinking water should therefore be of concern.A porcentagem de amostras positivas para Aeromonas foi de 12.7% para água mineral, 8.3% para água de poço artesiano e 6.5% para água do sistema público de abastecimento. O isolamento de Aeromonas spp. foi significativamente maior em água mineral e água de poço artesiano do que em água do sistema público. A ocorrência de Aeromonas spp. não teve correlação significativa com os indicadores de contaminação tradicionalmente utilizados (coliformes totais em amostras de água de poço artesiano. No entanto, esta correlação foi positiva e significativa em água mineral e água do sistema público. A presença ou ausência de correlação entre bactérias indicadoras e a presença de Aeromonas pode refletir o

Diversity and antibiotic resistance of Aeromonas spp. in drinking and waste water treatment plants.

Science.gov (United States)

Figueira, Vânia; Vaz-Moreira, Ivone; Silva, Márcia; Manaia, Célia M

2011-11-01

The taxonomic diversity and antibiotic resistance phenotypes of aeromonads were examined in samples from drinking and waste water treatment plants (surface, ground and disinfected water in a drinking water treatment plant, and raw and treated waste water) and tap water. Bacteria identification and intra-species variation were determined based on the analysis of the 16S rRNA, gyrB and cpn60 gene sequences. Resistance phenotypes were determined using the disc diffusion method. Aeromonas veronii prevailed in raw surface water, Aeromonas hydrophyla in ozonated water, and Aeromonas media and Aeromonas puntacta in waste water. No aeromonads were detected in ground water, after the chlorination tank or in tap water. Resistance to ceftazidime or meropenem was detected in isolates from the drinking water treatment plant and waste water isolates were intrinsically resistant to nalidixic acid. Most of the times, quinolone resistance was associated with the gyrA mutation in serine 83. The gene qnrS, but not the genes qnrA, B, C, D or qepA, was detected in both surface and waste water isolates. The gene aac(6')-ib-cr was detected in different waste water strains isolated in the presence of ciprofloxacin. Both quinolone resistance genes were detected only in the species A. media. This is the first study tracking antimicrobial resistance in aeromonads in drinking, tap and waste water and the importance of these bacteria as vectors of resistance in aquatic environments is discussed. Copyright © 2011 Elsevier Ltd. All rights reserved.

[Dermatophyte colonization on guinea pigs (Cavia porcellus) kept in pet stores. First report from Santiago, Chile].

Science.gov (United States)

Thomson, Pamela; Monsalves, Pamela; Maier, Liliana; Silva, Víctor

2015-01-01

Dermatophytes are pathogenic fungi that can be present in the flora of mammals, such as dogs, cats and rodents, which can be a source and transmission vehicle to other hosts, including humans. In Chile, there is a steady increase of acquiring guinea pigs (Cavia porcellus) as pets, with no local studies on their colonization by dermatophytes. To determine the presence of dermatophytes on clinically healthy guinea pigs, kept in pet stores in Santiago, Chile. A total of 52 clinically healthy animals were studied using the method by Mariat and Tapia (1966). The specimen culture and identification of the dermatophytes were performed using classical mycological procedures. Four guinea pigs (7.7%) out of 52 were colonized by dermatophytes, and were identified as Trichophyton mentagrophytes (3 cases) and Trichophyton verrucosum (one case). This study shows, for the first time in Chile, that guinea pigs can be colonized by dermatophytes, which should alert administrators of pet stores, veterinarians and physicians, to keep this in mind when purchasing or looking after this type of pet in a veterinary office. Copyright © 2013 Revista Iberoamericana de Micología. Published by Elsevier Espana. All rights reserved.

In vitro comparisons of the inhibitory activity of florfenicol copper sulfate and potassium permanganate towards Aeromonas hydrophila and Flavobacterium columnare

Science.gov (United States)

Aeromonas hydrophila and Flavobacterium columnare, the etiological agents of motile aeromonas septicemia (MAS) and columnaris disease, respectively, have been recently causing crippling moralities to the sunshine bass, Morone chrysops female X Morone saxatilis male (Percichthyidae), industry in the ...

Modular co-culture engineering, a new approach for metabolic engineering.

Science.gov (United States)

Zhang, Haoran; Wang, Xiaonan

2016-09-01

With the development of metabolic engineering, employment of a selected microbial host for accommodation of a designed biosynthetic pathway to produce a target compound has achieved tremendous success in the past several decades. Yet, increasing requirements for sophisticated microbial biosynthesis call for establishment and application of more advanced metabolic engineering methodologies. Recently, important progress has been made towards employing more than one engineered microbial strains to constitute synthetic co-cultures and modularizing the biosynthetic labor between the co-culture members in order to improve bioproduction performance. This emerging approach, referred to as modular co-culture engineering in this review, presents a valuable opportunity for expanding the scope of the broad field of metabolic engineering. We highlight representative research accomplishments using this approach, especially those utilizing metabolic engineering tools for microbial co-culture manipulation. Key benefits and major challenges associated with modular co-culture engineering are also presented and discussed. Copyright © 2016 International Metabolic Engineering Society. Published by Elsevier Inc. All rights reserved.

Investigation of Legionella Contamination in Bath Water Samples by Culture, Amoebic Co-Culture, and Real-Time Quantitative PCR Methods.

Science.gov (United States)

Edagawa, Akiko; Kimura, Akio; Kawabuchi-Kurata, Takako; Adachi, Shinichi; Furuhata, Katsunori; Miyamoto, Hiroshi

2015-10-19

We investigated Legionella contamination in bath water samples, collected from 68 bathing facilities in Japan, by culture, culture with amoebic co-culture, real-time quantitative PCR (qPCR), and real-time qPCR with amoebic co-culture. Using the conventional culture method, Legionella pneumophila was detected in 11 samples (11/68, 16.2%). Contrary to our expectation, the culture method with the amoebic co-culture technique did not increase the detection rate of Legionella (4/68, 5.9%). In contrast, a combination of the amoebic co-culture technique followed by qPCR successfully increased the detection rate (57/68, 83.8%) compared with real-time qPCR alone (46/68, 67.6%). Using real-time qPCR after culture with amoebic co-culture, more than 10-fold higher bacterial numbers were observed in 30 samples (30/68, 44.1%) compared with the same samples without co-culture. On the other hand, higher bacterial numbers were not observed after propagation by amoebae in 32 samples (32/68, 47.1%). Legionella was not detected in the remaining six samples (6/68, 8.8%), irrespective of the method. These results suggest that application of the amoebic co-culture technique prior to real-time qPCR may be useful for the sensitive detection of Legionella from bath water samples. Furthermore, a combination of amoebic co-culture and real-time qPCR might be useful to detect viable and virulent Legionella because their ability to invade and multiply within free-living amoebae is considered to correlate with their pathogenicity for humans. This is the first report evaluating the efficacy of the amoebic co-culture technique for detecting Legionella in bath water samples.

Immunological evaluation of Aeromonas infection in albino rats | El ...

African Journals Online (AJOL)

Aeromonas hydrophila is the most common bacterial species causing an acute self limiting diarrheal illness in mammals. Albino rats were categorized into treated and control groups. Treated groups were injected daily intraperitoneally with 0.2ml of bacterial suspension containing 6x106 cells/ml of 0.9% NaCl, while the ...

Unusual microorganisms and antimicrobial resistances in a group of Syrian migrants: Sentinel surveillance data from an asylum seekers centre in Italy.

Science.gov (United States)

Angeletti, Silvia; Ceccarelli, Giancarlo; Vita, Serena; Dicuonzo, Giordano; Lopalco, Maurizio; Dedej, Etleva; Blasi, Aletheia; Antonelli, Francesca; Conti, Alessia; De Cesaris, Marina; Farchi, Francesca; Lo Presti, Alessandra; Ciccozzi, Massimo

2016-01-01

Three years of civil war in Syria have caused death and increase of communicable diseases. The suffering population has been forced to migrate creating a fertile condition for epidemic spread of infection within the refugee camps. Forty-eight Syrian migrants, upon their arrival in Italy, were accommodated at the asylum seekers centre of Castelnuovo di Porto. They received a physical examination and were subjected to microbiological surveillance by blood, rectal, pharyngeal and nasal swabs collection and delivering to the Clinical Pathology and Microbiology Laboratory of the University Campus Bio-Medico of Rome. All refugees resulted negative for HBV, HCV and HIV infections. In swabs a large number of unusual gram-negative bacteria species were isolated, such as Pseudomonas putida, Pseudomonas monteilii, Pseudomonas fulva, Pseudomonas moselii, Aeromonas veronii, Aeromonas caviae, Aeromonas hydrophila, Acinteobacter guilloviae, Acinteobacter lowffii; Acinetobacter johnsonii; Acinteobacter tjernbergae; Pantoea agglomerans; Pantoea calida. Among isolates, strains resistant to carbapenems, ESBL producers and methicillin resistant were found. The microbiological surveillance performed represents a useful action to understand refugees health status and to trace unusual microorganisms movement even carriers of antimicrobial resistance during migrants traveling. Copyright © 2016 Elsevier Ltd. All rights reserved.

Investigation of Legionella Contamination in Bath Water Samples by Culture, Amoebic Co-Culture, and Real-Time Quantitative PCR Methods

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Akiko Edagawa

2015-10-01

Full Text Available We investigated Legionella contamination in bath water samples, collected from 68 bathing facilities in Japan, by culture, culture with amoebic co-culture, real-time quantitative PCR (qPCR, and real-time qPCR with amoebic co-culture. Using the conventional culture method, Legionella pneumophila was detected in 11 samples (11/68, 16.2%. Contrary to our expectation, the culture method with the amoebic co-culture technique did not increase the detection rate of Legionella (4/68, 5.9%. In contrast, a combination of the amoebic co-culture technique followed by qPCR successfully increased the detection rate (57/68, 83.8% compared with real-time qPCR alone (46/68, 67.6%. Using real-time qPCR after culture with amoebic co-culture, more than 10-fold higher bacterial numbers were observed in 30 samples (30/68, 44.1% compared with the same samples without co-culture. On the other hand, higher bacterial numbers were not observed after propagation by amoebae in 32 samples (32/68, 47.1%. Legionella was not detected in the remaining six samples (6/68, 8.8%, irrespective of the method. These results suggest that application of the amoebic co-culture technique prior to real-time qPCR may be useful for the sensitive detection of Legionella from bath water samples. Furthermore, a combination of amoebic co-culture and real-time qPCR might be useful to detect viable and virulent Legionella because their ability to invade and multiply within free-living amoebae is considered to correlate with their pathogenicity for humans. This is the first report evaluating the efficacy of the amoebic co-culture technique for detecting Legionella in bath water samples.

Potential of Chromolaena odorata Leaf as A Cure of Aeromonas hydrophila on Giant Gouramy (Osphronemus gouramy

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Y. Hadiroseyani

2007-07-01

Full Text Available This study was conducted to examine the potency of Chromolaena odorata leaf extract as a medicine for skin eruption disease caused by  Aeromonas hydrophila in giant gouramy Osphronemus gouramy.  Leaf extract of Chromolaena odorata for in vitro test was 0 (as control, 13000, 15000, 17000, 19000 and 21000 ppm, poured onto TSA medium containing bacteria 103 cfu/ml, and then is incubated for 24 hours. In vivo test was performed by injecting bacteria 0.1 ml of 109 cfu/ml intramuscularly into giant gouramy (14 g weight, and then  fish were maintained in the water containing 15000 ppm of Chromolaena odorata leaf extract. In vitro study showed that prevention area of leaf extract against Aeromonas hydrophila was increase by increasing the concentration of leaf extract used, reached 9,33 mm.  Prevention zone of leaf extract by difusion tends to constant, reached 7,6 mm. By in vivo test, survival rate of giant gouramy infected by Aeromonas hydrophila was no significantly different between dosages of leaf extract.  All treated fish, excluded control died after 24 hours infection. Keywords: Aeromonas hydrophila, Osphronemus gouramy, Chromolaena odorata   ABSTRAK Penelitian ini dilakukan untuk mengetahui potensi daun kirinyuh Chromolaena odorata sebagai obat untuk penyakit cacar yang diakibatkan oleh Aeromonas hydrophila pada ikan gurame Osphronemus gouramy. Konsentrasi ekstrak daun Chromolaena odorata untuk uji in vitro adalah 13000, 15000, 17000, 19000 dan 21000 serta 0 ppm sebagai kontrol, yang diletakkan di atas media TSA yang telah mengandung biakan bakteri 103 cfu/ml dan diinkubasi selama 24 jam. Uji in vivo dilakukan dengan menginjeksikan bakteri  sebanyak 0,1 ml (109 cfu/ml secara intramuskular ke ikan gurame (berat 14 g dan kemudian ikan dipelihara dalam air yang mengandung ekstrak daun kirinyuh 15000 ppm. Hasil uji in virto menunjukkan bahwa semakin tinggi konsentrasi ekstrak daun kirinyuh basah semakin efektif dalam menghambat perkembangan

Advances in tissue engineering through stem cell-based co-culture.

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Paschos, Nikolaos K; Brown, Wendy E; Eswaramoorthy, Rajalakshmanan; Hu, Jerry C; Athanasiou, Kyriacos A

2015-05-01

Stem cells are the future in tissue engineering and regeneration. In a co-culture, stem cells not only provide a target cell source with multipotent differentiation capacity, but can also act as assisting cells that promote tissue homeostasis, metabolism, growth and repair. Their incorporation into co-culture systems seems to be important in the creation of complex tissues or organs. In this review, critical aspects of stem cell use in co-culture systems are discussed. Direct and indirect co-culture methodologies used in tissue engineering are described, along with various characteristics of cellular interactions in these systems. Direct cell-cell contact, cell-extracellular matrix interaction and signalling via soluble factors are presented. The advantages of stem cell co-culture strategies and their applications in tissue engineering and regenerative medicine are portrayed through specific examples for several tissues, including orthopaedic soft tissues, bone, heart, vasculature, lung, kidney, liver and nerve. A concise review of the progress and the lessons learned are provided, with a focus on recent developments and their implications. It is hoped that knowledge developed from one tissue can be translated to other tissues. Finally, we address challenges in tissue engineering and regenerative medicine that can potentially be overcome via employing strategies for stem cell co-culture use. Copyright © 2014 John Wiley & Sons, Ltd.

A rapid MALDI-TOF MS identification database at genospecies level for clinical and environmental Aeromonas strains.

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Cinzia Benagli

Full Text Available The genus Aeromonas has undergone a number of taxonomic and nomenclature revisions over the past 20 years, and new (subspecies and biogroups are continuously described. Standard identification methods such as biochemical characterization have deficiencies and do not allow clarification of the taxonomic position. This report describes the development of a matrix-assisted laser desorption/ionisation-time of flight mass spectrometry (MALDI-TOF MS identification database for a rapid identification of clinical and environmental Aeromonas isolates.

Testicular Sertoli cells influence the proliferation and immunogenicity of co-cultured endothelial cells

International Nuclear Information System (INIS)

Fan, Ping; He, Lan; Pu, Dan; Lv, Xiaohong; Zhou, Wenxu; Sun, Yining; Hu, Nan

2011-01-01

Research highlights: → The proliferation of dramatic increased by co-cultured with Sertoli cells. → VEGF receptor-2 expression of ECs was up-regulated by co-cultured with Sertoli cells. → The MHC expression of ECs induced by INF-γ and IL-6, IL-8 and sICAM induced by TNF-α decreased respectively after co-cultured with Sertoli cells. → ECs co-cultured with Sertoli cells also didn't increase the stimulation index of spleen lymphocytes. -- Abstract: The major problem of the application of endothelial cells (ECs) in transplantation is the lack of proliferation and their immunogenicity. In this study, we co-cultured ECs with Sertoli cells to monitor whether Sertoli cells can influence the proliferation and immunogenicity of co-cultured ECs. Sertoli cells were isolated from adult testicular tissue. ECs were divided into the control group and the experimental group, which included three sub-groups co-cultured with 1 x 10 3 , 1 x 10 4 or 1 x 10 5 cell/ml of Sertoli cells. The growth and proliferation of ECs were observed microscopically, and the expression of vascular endothelial growth factor (VEGF) receptor-2 (KDR) was examined by Western blotting. In another experiment, ECs were divided into the control group, the single culture group and the co-culture group with the optimal concentration of Sertoli cells. After INF-γ and TNF-α were added to the culture medium, MHC II antigen expression was detected by immunofluorescence staining and western blotting; interleukin (IL)-6, IL-8 and soluble intercellular adhesion molecule (sICAM) were measured in the culture medium by ELISA. We demonstrated that 1 x 10 4 cell/ml Sertoli cells promoted the proliferation of co-cultured ECs more dramatically than that in other groups (P 4 cell/ml of the Sertoli cells was most effective in the up-regulation of KDR expression in the co-cultured ECs (P < 0.05). Sertoli cells can effectively suppress INF-γ-induced MHC II antigen expression in co-cultured ECs compared with single

Antibacterial efficacies of some plant extracts against Aeromonas and Pseudomonas diseases of farmed catfish (Heterobranchus longifilis

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Albert P. Ekanem

2011-11-01

Full Text Available Aeromonas and Pseudomonas diseases are responsible for mortalities of some farmed catfish in Nigeria. The objective of the study is to investigate the efficacies of extracts of some plants against Aeromonas and Pseudomonas diseases of Heterobranchus longifilis. Ethanol extracts of Phyllanthus amarus, Allium sativum, Artemisia annua, Citrus limon, Moringa oleifera, Allium cepa and Azadirachta indica were tested against Aeromonas hydrophila and Pseudomonas flourescens of H. longifilis by disc diffusion assay. Extracts of P. amarus, A. sativum, A. annua and C. limon were significantly (P<0.05 more sensitive to A. hydrophila and P. flourescens than M. oleifera, A. cepa and A. indica which were effective against P. flourescens. Minimum inhibitory concentrations (MIC of the extracts were 25mg/ml for P. amarus and A. annua; 25 and 100mg/ml for C. lemon and A. cepa respectively and 50mg/ml for A. indica.  Alkaloid was demonstrated in all plants except A. annua by qualitative methods. Moderate amount (++ of cardiac glycosides was demonstrated in A. sativum, M. oleifera and P. amarus. Saponin (+++ was present in M. oleifera and A. indica while, tannin (++ was present in M. oleifera, P. amarus and A. indica respectively. Phlobatanins and Anthraquinones (++ were present in P. amarus and M. oleifera respectively.  Extracts of aforementioned plants have potentials as therapy against Aeromonas hydrophila and Pseudomonas flourescens of farmed catfish.

Combined effect of gamma-irradiation and conventional cooking on Aeromonas hydrophila in meatball

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Ozbaş, Z. Y.; Vural, H.; Aytac, S. A.

1996-07-01

Irradiation combined with a conventional cooking procedure was applied to meatball and the effects on bacterial load and inoculated Aeromonas hydrophila were determined. Meatball samples were irradiated by using a 60Co source at the dose levels of 0, 0.30, 0.75, 1.50, 2.50 kGy and cold stored at 4 +/- 1 degrees C for 7 days. Bacterial load and the count of A. hydrophila decreased when the irradiation dose level increased. A minimum inhibition effect was found at the dose of 0.30 kGy. Irradiation in combination with a conventional cooking procedure was found to be more effective in reducing A. hydrophila and the bacterial load in meatball. This study indicated that a dose of 0.75 kGy was sufficient to destroy approximately 10(4) cfu/g of A. hydrophila in meatball.

Combined effect of gamma-irradiation and conventional cooking on Aeromonas hydrophila in meatball.

Science.gov (United States)

Ozbaş, Z Y; Vural, H; Aytaç, S A

1996-01-01

Irradiation combined with a conventional cooking procedure was applied to meatball and the effects on bacterial load and inoculated Aeromonas hydrophila were determined. Meatball samples were irradiated by using a 60Co source at the dose levels of 0, 0.30, 0.75, 1.50, 2.50 kGy and cold stored at 4 +/- 1 degrees C for 7 days. Bacterial load and the count of A. hydrophila decreased when the irradiation dose level increased. A minimum inhibition effect was found at the dose of 0.30 kGy. Irradiation in combination with a conventional cooking procedure was found to be more effective in reducing A. hydrophila and the bacterial load in meatball. This study indicated that a dose of 0.75 kGy was sufficient to destroy approximately 10(4) cfu/g of A. hydrophila in meatball.

Combined effect of gamma-irradiation and conventional cooking on Aeromonas hydrophila in meatball

International Nuclear Information System (INIS)

Ozbaş, Z.Y.; Vural, H.; Aytac, S.A.

1996-01-01

Irradiation combined with a conventional cooking procedure was applied to meatball and the effects on bacterial load and inoculated Aeromonas hydrophila were determined. Meatball samples were irradiated by using a 60Co source at the dose levels of 0, 0.30, 0.75, 1.50, 2.50 kGy and cold stored at 4 +/- 1 degrees C for 7 days. Bacterial load and the count of A. hydrophila decreased when the irradiation dose level increased. A minimum inhibition effect was found at the dose of 0.30 kGy. Irradiation in combination with a conventional cooking procedure was found to be more effective in reducing A. hydrophila and the bacterial load in meatball. This study indicated that a dose of 0.75 kGy was sufficient to destroy approximately 10(4) cfu/g of A. hydrophila in meatball

Characterization of atypical Aeromonas salmonicida isolates by ribotyping and plasmid profiling

DEFF Research Database (Denmark)

Pedersen, Karl; Dalsgaard, Inger; Larsen, J.L.

1996-01-01

A total of 38 strains of atypical Aeromonas salmonicida, three oxidase-negative but otherwise typical Aer. salmonicida, three typical Aer. salmonicida, and two reference strains, isolated from several countries and fish species were examined with respect to rRNA gene restriction patterns (ribotypes...

Waste cooking oil as substrate for biosynthesis of poly(3-hydroxybutyrate and poly(3-hydroxybutyrate-co-3-hydroxyhexanoate: Turning waste into a value-added product

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Yang, T. A.

2013-01-01

Full Text Available Aims: Improper disposal of domestic wastes, such as waste cooking oil (WCO, contributes to the deterioration of the environment and may lead to health problems. In this study, we evaluated the potential of plant-based WCO as a carbon source for the commercial biosynthesis of the bio-plastics, poly(3-hydroxybutyrate and poly(3-hydroxybutyrate-co-3-hydroxyhexanoate. The consumption of WCO for this purpose would mitigate their pollution of the environment at the same time.Methodology and Results: WCO collected from several cafeterias in USM was tested as the carbon source for polyhydroxyalkanoates (PHA production. A selection of suitable nitrogen source was first conducted in order to obtain an acceptable number of dry cell weight (DCW and PHA content. Urea was found to be a suitable nitrogen source for the two bacterial strains used in our study, Cupriavidus necator H16 and its transformed mutant, C. necator PHB¯4 harboring the PHA synthase gene of Aeromonas caviae (PHB¯4/pBBREE32d13. With WCO as the sole carbon source, C. necator H16 yielded a relatively good dry cell weight (DCW=25.4 g/L, with 71 wt% poly(3-hydroxybutyrate P(3HBcontent. In comparison, the DCW obtained with fresh cooking oil (FCO was 24.8 g/L. The production of poly(3 hydroxybutyrate-co-3- hydroxyhexanoate [P(3HB-co-3HHx] from WCO by the transformant C. necator PHB¯4 was comparable, yielding a DCW of 22.3 g/L and P(3HB-co-3HHx content of 85 wt%. Lipase activities for both bacterial strains reached a maximum after 72 h of cultivation when time profile was conducted. Conclusion, significance and impact of study: The use of WCO as a carbon source in the biosynthesis of the bioplastic, PHA, turns a polluting domestic waste into a value-added biodegradable product. This renewable source material can thus be exploited for the low cost production of PHA.

Aspects of constitutive and acquired antibioresistance in Aeromonas hydrophila strains isolated from water sources.

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Balotescu, Carmen; Israil, Anca; Radu, Roxana; Alexandru, Ionela; Dobre, Georgeta

2003-01-01

Over the last three decades, the literature pointed out the implications of Aeromonas species in human pathology. These species were described as being involved in intestinal (several outbreaks of acute gastroenteritis of choleric/dysenteric form or chronic diarrhoea, ulcerative colitis, etc.) in normal adults or children, as well as in extraintestinal infections in immunocompromised hosts. This last aspect included a large range of cutaneous injuries (micronecrosis, abscesses, bums, cellulites, furunculosis), joint, bones, respiratory, urinary tract, ocular infections up to meningitis, endocarditis, peritonitis, hepatobilliary disease, endotoxic shock and septicemia (as consequence of leech microvascular surgery). During the last decade, the literature reported a high mortality in Aeromonas infections determined by certain phenospecies (A. hydrophila and A. veronii) especially in extraintestinal infections in immunocompromised patients. In microbiologists' opinion this high rate of mortality was probably due to poor knowledge concerning the aspects of antibioresistance in Aeromonas strains, to empiric treatments with antibiotics to which these bacteria exhibiting constitutive resistance lead to insuccessful results, and at last to the increasing trend of aeromonads resistance to certain antibiotics after 1996. The literature mentioned also that for a great number of Beta-lactamase producing Aeromonas strains, the use of microdilution method (by comparison to disk diffusion in agar medium) giving false results made more difficult the true knowledge of Aeromonas antibioresistance patterns. At the same time, in 2002, the literature mentioned 4 ecological compartments considered as "reservoirs for dissemination and transfer of microbial antibioresistance i.e. humans, animals, plants and natural soil and water. In the last time, more and more data of the literature revealed that some bacteria with role of reservoir of antibioresistance in the natural environment, even

Draft genome sequences of three Aeromonas hybrophila isolates from catfish and tilapia

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Aeromonas hydrophila is a Gram-negative bacteria that is particularly adapted to freshwater environments and can cause severe infections in fish and humans. Here we report the draft genomes of three A. hydrophila catfish and tilapia isolates....

Necrotizing Fasciitis Secondary to Aeromonas Infection Presenting with Septic Shock

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Nikhil Bhatia

2017-01-01

Full Text Available This report describes a case of necrotizing fasciitis presenting with septic shock due to an Aeromonas infection. The patient cut his foot while mowing the lawn and then spent time in a pool with black mold. He began feeling ill and developed swelling and a quarter-sized black area on his right lower extremity. Despite being hemodynamically unstable with systolic blood pressure in the low 70s, the patient was transferred to our facility from outside hospital 100 miles away. Upon arriving to facility, the patient appeared to be septic and the infected area of skin had grown. Irrigation and debridement were performed and appropriate antibiotic therapy was given; however, the patient subsequently died on hospital day 8. On review of the literature, cases of necrotizing fasciitis due to Aeromonas infection have been treated successfully with the aforementioned therapy; however, there is high mortality associated with these infections, many times related to a delayed diagnosis. Our patient also had multiple poor prognostic factors including hepatic dysfunction and immunosuppression.

[Research progress of co-culture system for constructing vascularized tissue engineered bone].

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Fu, Weili; Xiang, Zhou

2014-02-01

To review the research progress of the co-culture system for constructing vascularized tissue engineered bone. The recent literature concerning the co-culture system for constructing vascularized tissue engineered bone was reviewed, including the selection of osteogenic and endothelial lineages, the design and surface modification of scaffolds, the models and dimensions of the co-culture system, the mechanism, the culture conditions, and their application progress. The construction of vascularized tissue engineered bone is the prerequisite for their survival and further clinical application in vivo. Mesenchymal stem cells (owning the excellent osteogenic potential) and endothelial progenitor cells (capable of directional differentiation into endothelial cell) are considered as attractive cell types for the co-culture system to construct vascularized tissue engineered bone. The culture conditions need to be further optimized. Furthermore, how to achieve the clinical goals of minimal invasion and autologous transplantation also need to be further studied. The strategy of the co-culture system for constructing vascularized tissue engineered bone would have a very broad prospects for clinical application in future.

Antibiogram, Adhesive Characteristics, and Incidence of Class 1 Integron in Aeromonas Species Isolated from Two South African Rivers

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Isoken H. Igbinosa

2013-01-01

Full Text Available Aeromonas species are well distributed in freshwater environments, and their natural susceptibility to antimicrobials renders them interesting candidates for the survey of antimicrobial resistance in freshwater milieu. Water samples were collected from Kat and Tyume rivers in the Eastern Cape province of South Africa, and a total of 45 isolates identified as Aeromonas species were recovered from the two rivers. All Aeromonas isolates were resistant to oxacillin, penicillin, clindamycin, cephalothin, vancomycin, and rifamycin, while appreciable susceptibilities (89.3 : 94.1%, 82.1 : 94.1%, 85.7 : 88.2%, and 92.9 : 88.2% were observed against ciprofloxacin, chloramphenicol, nitrofurantoin, and gentamicin from Kat and Tyume rivers, respectively. Multiple antibiotic resistance (MAR indices ranged from 0.016 to 0.044 for the two rivers. Class 1 integron was detected in about 20% of the isolates, and all the isolates except one showed ability to produce biofilm in vitro as weak producers (53.33%, moderate producers (15.56%, and strong producers (28.9%. This investigation provides a baseline data on antibiotic resistance as well as the adhesive characteristics of Aeromonas isolates from Tyume and Kat rivers in the Eastern Cape province of South Africa.

Application of a continuously stirred tank bioreactor (CSTR) for bioremediation of hydrocarbon-rich industrial wastewater effluents.

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Gargouri, Boutheina; Karray, Fatma; Mhiri, Najla; Aloui, Fathi; Sayadi, Sami

2011-05-15

A continuously stirred tank bioreactor (CSTR) was used to optimize feasible and reliable bioprocess system in order to treat hydrocarbon-rich industrial wastewaters. A successful bioremediation was developed by an efficient acclimatized microbial consortium. After an experimental period of 225 days, the process was shown to be highly efficient in decontaminating the wastewater. The performance of the bioaugmented reactor was demonstrated by the reduction of COD rates up to 95%. The residual total petroleum hydrocarbon (TPH) decreased from 320 mg TPH l(-1) to 8 mg TPH l(-1). Analysis using gas chromatography-mass spectrometry (GC-MS) identified 26 hydrocarbons. The use of the mixed cultures demonstrated high degradation performance for hydrocarbons range n-alkanes (C10-C35). Six microbial isolates from the CSTR were characterized and species identification was confirmed by sequencing the 16S rRNA genes. The partial 16S rRNA gene sequences demonstrated that 5 strains were closely related to Aeromonas punctata (Aeromonas caviae), Bacillus cereus, Ochrobactrum intermedium, Stenotrophomonas maltophilia and Rhodococcus sp. The 6th isolate was affiliated to genera Achromobacter. Besides, the treated wastewater could be considered as non toxic according to the phytotoxicity test since the germination index of Lepidium sativum ranged between 57 and 95%. The treatment provided satisfactory results and presents a feasible technology for the treatment of hydrocarbon-rich wastewater from petrochemical industries and petroleum refineries. Copyright © 2011 Elsevier B.V. All rights reserved.

Effect of salinity and incubation time of planktonic cells on biofilm formation, motility, exoprotease production, and quorum sensing of Aeromonas hydrophila.

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Jahid, Iqbal Kabir; Mizan, Md Furkanur Rahaman; Ha, Angela J; Ha, Sang-Do

2015-08-01

The aim of this study was to determine the effect of salinity and age of cultures on quorum sensing, exoprotease production, and biofilm formation by Aeromonas hydrophila on stainless steel (SS) and crab shell as substrates. Biofilm formation was assessed at various salinities, from fresh (0%) to saline water (3.0%). For young and old cultures, planktonic cells were grown at 30 °C for 24 h and 96 h, respectively. Biofilm formation was assessed on SS, glass, and crab shell; viable counts were determined in R2A agar for SS and glass, but Aeromonas-selective media was used for crab shell samples to eliminate bacterial contamination. Exoprotease activity was assessed using a Fluoro™ protease assay kit. Quantification of acyl-homoserine lactone (AHL) was performed using the bioreporter strain Chromobacterium violaceum CV026 and the concentration was confirmed using high-performance liquid chromatography (HPLC). The concentration of autoinducer-2 (AI-2) was determined with Vibrio harveyi BB170. The biofilm structure at various salinities (0-3 %) was assessed using field emission electron microscopy (FESEM). Young cultures of A. hydrophila grown at 0-0.25% salinity showed gradual increasing of biofilm formation on SS, glass and crab shell; swarming and swimming motility; exoproteases production, AHL and AI-2 quorum sensing; while all these phenotypic characters reduced from 0.5 to 3.0% salinity. The FESEM images also showed that from 0 to 0.25% salinity stimulated formation of three-dimensional biofilm structures that also broke through the surface by utilizing the chitin surfaces of crab, while 3% salinity stimulated attachment only for young cultures. However, in marked contrast, salinity (0.1-3%) had no effect on the stimulation of biofilm formation or on phenotypic characters for old cultures. However, all concentrations reduced biofilm formation, motility, protease production and quorum sensing for old culture. Overall, 0-0.25% salinity enhanced biofilm formation

An investigation of the bactericidal activity of selected essential oils to Aeromonas spp.

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Clifford E. Starliper

2015-01-01

Full Text Available Diseases of fishes caused by Aeromonas spp. are common, have broad host ranges and may cause high mortality. Treatments of captive-reared populations using antimicrobials are limited with concerns for bacterial resistance development and environmental dissemination. This study was done to determine whether selected plant-derived essential oils were bactericidal to Aeromonas spp. Initially, twelve essential oils were evaluated using a disk diffusion assay to an isolate of A. salmonicida subsp. salmonicida, cause of fish furunculosis. The greatest zones of inhibition were obtained with oils of cinnamon Cinnamomum cassia, oregano Origanum vulgare, lemongrass Cymbopogon citratus and thyme Thymus vulgaris. Minimum bactericidal concentrations (MBC’s were determined for these four oils, Allimed® (garlic extract, Allium sativum and colloidal silver to sixty-nine isolates representing nine Aeromonas spp. The lowest mean MBCs (0.02–0.04% were obtained with three different sources of cinnamon oil. MBCs for three sources of oregano and lemongrass oils ranged from 0.14% to 0.30% and 0.10% to 0.65%, respectively, and for two thyme oils were 2.11% and 2.22%. The highest concentration (5% of Allimed® tested resulted in MBCs to twelve isolates. A concentration of silver greater than 15 mg/L would be required to determine MBCs for all but one isolate.

[Role of Aeromonas in the monitoring of a hydroecosystem in the Volga-Caspian area].

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Obukhova, O V; Lartseva, L V; Lisitskaia, I A

2011-01-01

The paper gives data obtained over many years on the quantity of bacteria of the genus Aeromonas in the hydroecosystems of the delta of the Volga and Caspian Sea and in commercial fishes (sturgeon (Acipenseridae), pike perch (Stizostedion), carp (Cyprinus carpio), silurus (Silurus glanis), etc.). It provides a detailed analysis of their seasonal trends and spatial distribution, markers of their pathogenicity, and antibiotic resistance. The findings suggest that Aeromonas is a causative agent of sapronoses and meets the criteria for natural focal infections. They show a wide range of tolerance to the abiotic factors of hydroecosystems, as well as mixotrophy and autrophic nutrition in the environment and they are of great sanitary and hygienic importance.

Application of cell co-culture system to study fat and muscle cells.

Science.gov (United States)

Pandurangan, Muthuraman; Hwang, Inho

2014-09-01

Animal cell culture is a highly complex process, in which cells are grown under specific conditions. The growth and development of these cells is a highly unnatural process in vitro condition. Cells are removed from animal tissues and artificially cultured in various culture vessels. Vitamins, minerals, and serum growth factors are supplied to maintain cell viability. Obtaining result homogeneity of in vitro and in vivo experiments is rare, because their structure and function are different. Living tissues have highly ordered complex architecture and are three-dimensional (3D) in structure. The interaction between adjacent cell types is quite distinct from the in vitro cell culture, which is usually two-dimensional (2D). Co-culture systems are studied to analyze the interactions between the two different cell types. The muscle and fat co-culture system is useful in addressing several questions related to muscle modeling, muscle degeneration, apoptosis, and muscle regeneration. Co-culture of C2C12 and 3T3-L1 cells could be a useful diagnostic tool to understand the muscle and fat formation in animals. Even though, co-culture systems have certain limitations, they provide a more realistic 3D view and information than the individual cell culture system. It is suggested that co-culture systems are useful in evaluating the intercellular communication and composition of two different cell types.

Optimization of the Static Human Osteoblast/Osteoclast Co-culture System

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James Jam Jolly

2018-03-01

Full Text Available Osteoblasts (OBs and osteoclasts (OCs are 2 major groups of bone cells. Their cell-to-cell interactions are important to ensure the continuity of the bone-remodeling process. Therefore, the present study was carried out to optimize an OB/OC co-culture system utilizing the human OB cell line hFOB 1.19 and OCs extracted from peripheral blood mononuclear cells (PBMNCs. It was a 2-step procedure, involving the optimization of the OB culture and the co-culture of the OBs with PBMNCs at an optimum ratio. Firstly, pre-OBs were cultured to 90% confluency and the time required for differentiation was determined. OB differentiation was determined using the van Gieson staining to detect the presence of collagen and Alizarin Red for calcium. Secondly, OBs and OCs were co-cultured at the ratios of 1 OC: 1 OB, 1 OC: 4 OBs, 2 OCs: 1 OB, and 1 OC: 2 OBs. Tartrate-resistant acid phosphatase (TRAP staining was used to detect the differentiation of the OCs. The results showed that collagen was present on day 1, whereas calcium was detected as early as day 3. Based on the result of TRAP staining, 1 OC: 2 OBs was taken as the most appropriate ratio. No macrophage colony-stimulating factor and receptor activator of the nuclear factor-κB ligand were added because they were provided by the OBs. In conclusion, these optimization processes are vital as they ensure the exact time point and ratio of the OB/OC co-culture in order to produce a reliable and reproducible co-culture system.

Spirulina chitosan gel induction on healing process of Cavia cobaya post extraction socket

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Rostiny Rostiny

2014-03-01

Full Text Available Background: Prominent residual ridge is necessary to gain retention and stabilility for succesful prosthodontic treatment such as removable, fixed or implant. Spirulina is a natural substance that can help tissue healing and chitosan also a natural substance that reported to have the ability to help bone remodelling. The combination gel of spirulina and chitosan could be considered as an alternative material to maintain residual ridge height after tooth extraction. Purpose: The aim of study was to examine the effect of combination gel of Spirulina and chitosan on healing process of Cavia cobaya post tooth extraction socket by counting the amount of osteoclast, osteoblast and colagen as an indicator. Methods: Twenty eight cavia cobaya were divided into 4 groups. Insisive mandible extraction was done and the sockets were filled with 3% CMCNa for control groups, 3% spirulina chitosan 200 mg for group 1, 6% spirulina chitosan 200 mg for group 2, 12% spirulina chitosan 200 mg for group 3. After 30 days, histopathology examination was done by using microscope to count the amount of osteoclast, osteoblast and collagen. Results: Data was analyzed by using Anova and Tukey HSD. For osteoclast, there was no significant different between every groups, while for osteoblast and collagen there was significant different between groups. The results showed that induction of combination gel spirulina chitosan was able to accumulate collagen fiber and resulting faster wound healing. Conclusion: Combination 12% gel spirulina chitosan 200 mg could be used as an alternative material for better bone remodeling after tooth extraction.Latar belakang: Residual ridge yang prominen sangat dibutuhkan untuk mendapatkan retensi dan stabilitas untuk menunjang keberhasilan perawatan di bidang prostodonsia seperti pada kasus removable, fixed atau implant. Tindakan pencabutan gigi dapat merusak jaringan periodontal, sementum dan tulang alveolar yang mengakibatkan resorbsi ridge

Recovery of atypical Aeromonas salmonicida from ulcerated fish from the Baltic Sea

DEFF Research Database (Denmark)

Wiklund, T.; Tabolina, I; Bezgachina, T.V.

1999-01-01

Ulcerated fish of six different species were collected during the BMB/ICES Sea-going Workshop "Fish Diseases and Parasites in the Baltic Sea", 25 November to 8 December 1994, and examined for bacteriological infections. Atypical Aeromonas salmonicida strains were isolated from the majority...

Engraftment Outcomes after HPC Co-Culture with Mesenchymal Stromal Cells and Osteoblasts

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Matthew M. Cook

2013-09-01

Full Text Available Haematopoietic stem cell (HSC transplantation is an established cell-based therapy for a number of haematological diseases. To enhance this therapy, there is considerable interest in expanding HSCs in artificial niches prior to transplantation. This study compared murine HSC expansion supported through co-culture on monolayers of either undifferentiated mesenchymal stromal cells (MSCs or osteoblasts. Sorted Lineage− Sca-1+ c-kit+ (LSK haematopoietic stem/progenitor cells (HPC demonstrated proliferative capacity on both stromal monolayers with the greatest expansion of LSK shown in cultures supported by osteoblast monolayers. After transplantation, both types of bulk-expanded cultures were capable of engrafting and repopulating lethally irradiated primary and secondary murine recipients. LSKs co-cultured on MSCs showed comparable, but not superior, reconstitution ability to that of freshly isolated LSKs. Surprisingly, however, osteoblast co-cultured LSKs showed significantly poorer haematopoietic reconstitution compared to LSKs co-cultured on MSCs, likely due to a delay in short-term reconstitution. We demonstrated that stromal monolayers can be used to maintain, but not expand, functional HSCs without a need for additional haematopoietic growth factors. We also demonstrated that despite apparently superior in vitro performance, co-injection of bulk cultures of osteoblasts and LSKs in vivo was detrimental to recipient survival and should be avoided in translation to clinical practice.

Transmissão do Trypanosoma cruzi em três gerações de Cavia porcellus sem a participação de triatomíneos

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Ítalo A. Sherlock

1976-02-01

Full Text Available Foi verificada a transmissão sucessiva do T. cruzi em três gerações da cobaia Cavia porcellus sem a participação de triatomíneos. Embora não fosse determinado qual das vias, se placentária, leite, excreções ou contágio direto pelo qual o protozoário foi transmitido para os descendentes, chama-se atenção para a importância da manutenção de reservatórios da Doença de Chagas, mesmo na ausência de vetores invertebrados.The sucessive transmission of Trypanosoma cruzi among three generations of the guinea pig Cavia porcellus without the participation of triatomine bugs is verified. Although the mode of transmission, such as congenital, infected milk or other excretion or direct contagion was not defined, this maybe of importance in natural maintenance of reservoirs of T. cruzi without the invertebrate vectors.

Draft Genome Sequence of Fish Pathogen Aeromonas bestiarum GA97-22.

Science.gov (United States)

Kumru, Salih; Tekedar, Hasan C; Griffin, Matt J; Waldbieser, Geoffrey C; Liles, Mark R; Sonstegard, Tad; Schroeder, Steven G; Lawrence, Mark L; Karsi, Attila

2018-06-14

Aeromonas bestiarum is a Gram-negative mesophilic motile bacterium causing acute hemorrhagic septicemia or chronic skin ulcers in fish. Here, we report the draft genome sequence of A. bestiarum strain GA97-22, which was isolated from rainbow trout in 1997. This genome sequence will improve our understanding of the complex taxonomy of motile aeromonads.

Detection limits of Legionella pneumophila in environmental samples after co-culture with Acanthamoeba polyphaga

Science.gov (United States)

2013-01-01

Background The efficiency of recovery and the detection limit of Legionella after co-culture with Acanthamoeba polyphaga are not known and so far no investigations have been carried out to determine the efficiency of the recovery of Legionella spp. by co-culture and compare it with that of conventional culturing methods. This study aimed to assess the detection limits of co-culture compared to culture for Legionella pneumophila in compost and air samples. Compost and air samples were spiked with known concentrations of L. pneumophila. Direct culturing and co-culture with amoebae were used in parallel to isolate L. pneumophila and recovery standard curves for both methods were produced for each sample. Results The co-culture proved to be more sensitive than the reference method, detecting 102-103 L. pneumophila cells in 1 g of spiked compost or 1 m3 of spiked air, as compared to 105-106 cells in 1 g of spiked compost and 1 m3 of spiked air. Conclusions Co-culture with amoebae is a useful, sensitive and reliable technique to enrich L. pneumophila in environmental samples that contain only low amounts of bacterial cells. PMID:23442526

Co-culture systems and technologies: taking synthetic biology to the next level.

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Goers, Lisa; Freemont, Paul; Polizzi, Karen M

2014-07-06

Co-culture techniques find myriad applications in biology for studying natural or synthetic interactions between cell populations. Such techniques are of great importance in synthetic biology, as multi-species cell consortia and other natural or synthetic ecology systems are widely seen to hold enormous potential for foundational research as well as novel industrial, medical and environmental applications with many proof-of-principle studies in recent years. What is needed for co-cultures to fulfil their potential? Cell-cell interactions in co-cultures are strongly influenced by the extracellular environment, which is determined by the experimental set-up, which therefore needs to be given careful consideration. An overview of existing experimental and theoretical co-culture set-ups in synthetic biology and adjacent fields is given here, and challenges and opportunities involved in such experiments are discussed. Greater focus on foundational technology developments for co-cultures is needed for many synthetic biology systems to realize their potential in both applications and answering biological questions. © 2014 The Author(s) Published by the Royal Society. All rights reserved.

Resistensi Bakteri Aeromonas sp. Isolat Ikan Piranha (Pygosentrus nattereri Bali Safari And Marine Park terhadap Antibiotik

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Ni Made Herawati

2013-11-01

Full Text Available Normal 0 false false false EN-US X-NONE X-NONE Piranha (Pygosentrus nattereri is a fish that has been conserved at Conservation Center of Bali Safari and Marine Park. In May 2012 there was a case that twelve Piranhas suffered from illness and two of them died without any specific clinical signs. The dead Piranhas were necropsied and the tissues or parts of organs were collected for microbial examination interests. Spesies specific bacterial examination was also included using Microgen GNA TM + B-ID system test The bacterial examination found that the fishes were infected by Aeromonas hydropila and Aeromonas veronii. Further examination about antibiotic resistance tests showed that those Aeromonas spp were resistant to chloramphenicol and a combination of trimethoprim and sulphamethoxazole. It can be suggested that, next, the usage of other antibiotics is strongly recommended for the case of Piranha infected by Aeromonas spp at Bali Safari and Marine Park. /* Style Definitions */ table.MsoNormalTable {mso-style-name:"Table Normal"; mso-tstyle-rowband-size:0; mso-tstyle-colband-size:0; mso-style-noshow:yes; mso-style-priority:99; mso-style-qformat:yes; mso-style-parent:""; mso-padding-alt:0in 5.4pt 0in 5.4pt; mso-para-margin:0in; mso-para-margin-bottom:.0001pt; text-align:justify; line-height:150%; mso-pagination:widow-orphan; font-size:11.0pt; font-family:"Calibri","sans-serif"; mso-ascii-font-family:Calibri; mso-ascii-theme-font:minor-latin; mso-fareast-font-family:"Times New Roman"; mso-fareast-theme-font:minor-fareast; mso-hansi-font-family:Calibri; mso-hansi-theme-font:minor-latin;}

Immobilized anaerobic fermentation for bio-fuel production by Clostridium co-culture.

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Xu, Lei; Tschirner, Ulrike

2014-08-01

Clostridium thermocellum/Clostridium thermolacticum co-culture fermentation has been shown to be a promising way of producing ethanol from several carbohydrates. In this research, immobilization techniques using sodium alginate and alkali pretreatment were successfully applied on this co-culture to improve the bio-ethanol fermentation performance during consolidated bio-processing (CBP). The ethanol yield obtained increased by over 60 % (as a percentage of the theoretical maximum) as compared to free cell fermentation. For cellobiose under optimized conditions, the ethanol yields were approaching about 85 % of the theoretical efficiency. To examine the feasibility of this immobilization co-culture on lignocellulosic biomass conversion, untreated and pretreated aspen biomasses were also used for fermentation experiments. The immobilized co-culture shows clear benefits in bio-ethanol production in the CBP process using pretreated aspen. With a 3-h, 9 % NaOH pretreatment, the aspen powder fermentation yields approached 78 % of the maximum theoretical efficiency, which is almost twice the yield of the untreated aspen fermentation.

Co-culture systems-based strategies for articular cartilage tissue engineering.

Science.gov (United States)

Zhang, Yu; Guo, Weimin; Wang, Mingjie; Hao, Chunxiang; Lu, Liang; Gao, Shuang; Zhang, Xueliang; Li, Xu; Chen, Mingxue; Li, Penghao; Jiang, Peng; Lu, Shibi; Liu, Shuyun; Guo, Quanyi

2018-03-01

Cartilage engineering facilitates repair and regeneration of damaged cartilage using engineered tissue that restores the functional properties of the impaired joint. The seed cells used most frequently in tissue engineering, are chondrocytes and mesenchymal stem cells. Seed cells activity plays a key role in the regeneration of functional cartilage tissue. However, seed cells undergo undesirable changes after in vitro processing procedures, such as degeneration of cartilage cells and induced hypertrophy of mesenchymal stem cells, which hinder cartilage tissue engineering. Compared to monoculture, which does not mimic the in vivo cellular environment, co-culture technology provides a more realistic microenvironment in terms of various physical, chemical, and biological factors. Co-culture technology is used in cartilage tissue engineering to overcome obstacles related to the degeneration of seed cells, and shows promise for cartilage regeneration and repair. In this review, we focus first on existing co-culture systems for cartilage tissue engineering and related fields, and discuss the conditions and mechanisms thereof. This is followed by methods for optimizing seed cell co-culture conditions to generate functional neo-cartilage tissue, which will lead to a new era in cartilage tissue engineering. © 2017 Wiley Periodicals, Inc.

The influence of Aloe vera and xenograft XCB toward of bone morpho protein 2 BMP2 expression and amount of osteoblast of alveolar bone induced into tooth extraction sockets Cavia cobaya

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Utari Kresnoadi

2014-12-01

Full Text Available Tooth extraction can cause inflammation leading to alveolar ridge resorption. In addition, prominent ridge has crucial role for making denture su-ccessfully. Thus, socket preservation is needed to prevent greater alveolar ridge resorption. An innovative material, a combination of Aloe vera and xe-nograft (XCB, is then considered as a biogenic stimulator that can reduce inflammation, as a result, the growth of alveolar bone is expected to be impro-ved. This research is aimed to prove whether the mixture of Aloe vera and xenograft can stimulate BMP2 and increase osteoblasts. Forty-eight Cavia co-baya animals were divided into eight groups each of which consisted of six animals. The mandibular incisors of those Cavia cobaya animals were then extracted and filled with PEG as Group Control, XCB as Group XCB, Aloe vera as Group Aloe vera, and a combination of Aloe vera +XCB as Group Aloe vera +XCB. Next, the first four groups were sacrificed seven days after extraction, and the second four groups were sacrificed 30 days after extrac-tion. And then, immunohistochemical and histopathology examinations were conducted to examine BMP2 expression and osteoblasts. Based on the re-sult known that the mixture of Aloe vera and xenograft can increase BMP2 expression and amount of osteoblasts. It can be concluded that the mixture of Aloe vera and xenograft can increase BMP2 expression and amount of osteoblast cel . It can be used as an alternative material to increase the growth of alveolar bone after extraction.

Biomaterials in co-culture systems: towards optimizing tissue integration and cell signaling within scaffolds.

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Battiston, Kyle G; Cheung, Jane W C; Jain, Devika; Santerre, J Paul

2014-05-01

Most natural tissues consist of multi-cellular systems made up of two or more cell types. However, some of these tissues may not regenerate themselves following tissue injury or disease without some form of intervention, such as from the use of tissue engineered constructs. Recent studies have increasingly used co-cultures in tissue engineering applications as these systems better model the natural tissues, both physically and biologically. This review aims to identify the challenges of using co-culture systems and to highlight different approaches with respect to the use of biomaterials in the use of such systems. The application of co-culture systems to stimulate a desired biological response and examples of studies within particular tissue engineering disciplines are summarized. A description of different analytical co-culture systems is also discussed and the role of biomaterials in the future of co-culture research are elaborated on. Understanding the complex cell-cell and cell-biomaterial interactions involved in co-culture systems will ultimately lead the field towards biomaterial concepts and designs with specific biochemical, electrical, and mechanical characteristics that are tailored towards the needs of distinct co-culture systems. Copyright © 2014 Elsevier Ltd. All rights reserved.

A new karyotype for the genus Cavia from a southern island of Brazil (Rodentia - Caviidae

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A. Gava

1998-03-01

Full Text Available Intraspecific karyotype variation in mammal species is very common and often caused by centromeric fusion of acrocentric chromosomes. We describe here a new karyotype 2n = 62 (FN = 112 for the genus Cavia from the Moleques do Sul Islands, of the southern coast of Brazil. We analyzed two male and four female karyotypes that had twenty-four biarmed pairs and six pairs of acrocentric chromosomes. The sexual pair consisted of a metacentric X-chromosome and a large acrocentric Y. C-bands were found in the centromeric and pericentromeric regions of almost all chromosomes, except for some small biarmed and acrocentric ones. Nucleolus organizer regions appeared in two biarmed chromosomes, and G-banding patterns were also seen.RESUMO A variação cariotípica nas espécies de mamíferos é bastante comum e geralmente causada pela fusão de cromossomos acrocêntricos. Foi descrito neste trabalho um novo cariótipo, com 2n = 62 e FN = 112, para o gênero Cavia proveniente das ilhas Moleques do Sul, da costa sul do Brasil. Foram analisados os cariótipos de dois machos e quatro fêmeas que possuiam 24 pares de cromossomos com dois braços e seis pares de acrocêntricos. O par sexual era constituído por um cromossomo X metacêntrico grande e um Y acrocêntrico. As bandas C estavam localizadas nas regiões centroméricas e pericentroméricas da maioria dos cromossomos, com exceção de alguns acrocêntricos e os cromossomos de dois braços menores. As regiões organizadoras de nucléolo ocorreram em dois cromossomos com dois braços e o padrão de bandamento G foi também apresentado.

Characterization of class 1 integrons associated with R-plasmids in clinical Aeromonas salmonicida isolates from various geographical areas

DEFF Research Database (Denmark)

Schmidt, A.S.; Bruun, Morten Sichlau; Larsen, J.L.

2001-01-01

Class 1 integrons were found in 26 of 40 antibiotic-resistant isolates of the fish pathogen Aeromonas salmonicida from Northern Europe and North America. Three different dhfr genes, conferring trimethoprim resistance, and one ant(3 " )1a aminoglycoside resistance gene were identified as gene...... inserts. The gene cassettes tended to be conserved among isolates from a particular geographical area. Nineteen isolates transferred R- plasmids carrying different tet determinants to Escherichia coli in filter mating assays, and in 15 cases, the class 1 integrons were co-transferred. Transferable...... sulphadiazine, trimethoprim and streptomycin resistances were invariably encoded by integrons. It thus appears that integron-encoded antibiotic resistance genes contribute substantially to the horizontal spread of antimicrobial resistance within this species, being associated with conjugative plasmids....

Ureolytic/Non-Ureolytic Bacteria Co-Cultured Self-Healing Agent for Cementitious Materials Crack Repair

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Hyeong Min Son

2018-05-01

Full Text Available The present study investigated the CaCO3 precipitation performance of ureolytic and non-ureolytic bacteria co-cultured as a self-healing agent for cementitious materials crack repair. Three different inoculum ratios of ureolytic Sporosarcina pasteurii and non-ureolytic Bacillus thuringiensis (10:0, 8:2, or 5:5 were used. The effect of coculturing ureolytic and non-ureolytic bacteria on microbial metabolism was investigated by measuring the rate of growth in urea-containing medium and the rate of NH4+ and CaCO3 production in urea–calcium lactate medium. The self-healing efficiency of co-cultured bacteria was examined by exposing cement mortar specimens with predefined cracks to media containing single urease-producing or co-cultured bacteria. The obtained results provide new findings, where CaCO3 precipitation is improved by co-culturing ureolytic and non-ureolytic bacteria, owing to the relatively faster growth rate of non-ureolytic bacteria. The crack filling rate correlated with the width of crack, in particular, specimens with a smaller crack width showed the faster filling effect, indicating that the crack width can be a dominant factor influencing the CaCO3 precipitation capacity of co-cultured bacteria.

Estereología Comparativa entre el Bazo del Cuye (Cavia porcellus) y la Rata (Rattus novergicus, Sprague Dawley)

OpenAIRE

Furrianca, María Cristina; Vásquez, Bélgica; del Sol, Mariano

2008-01-01

El bazo es el órgano linfático periférico más grande del organismo y conocer sus aspectos morfológicos cuantitativos es importante para determinar posibles patologías. El objetivo del estudio fue determinar en dos especies: cuye (Cavia porcellus) y rata (Rattus novergicus Sprague Dawley), las características estereológicas del bazo, para obtener patrones de normalidad cuantitativos, los que servirán de base para futuros estudios morfofuncionales. Se utilizaron 5 bazos de cada especie, clínica...

Functional 3-D cardiac co-culture model using bioactive chitosan nanofiber scaffolds.

Science.gov (United States)

Hussain, Ali; Collins, George; Yip, Derek; Cho, Cheul H

2013-02-01

The in vitro generation of a three-dimensional (3-D) myocardial tissue-like construct employing cells, biomaterials, and biomolecules is a promising strategy in cardiac tissue regeneration, drug testing, and tissue engineering applications. Despite significant progress in this field, current cardiac tissue models are not yet able to stably maintain functional characteristics of cardiomyocytes for long-term culture and therapeutic purposes. The objective of this study was to fabricate bioactive 3-D chitosan nanofiber scaffolds using an electrospinning technique and exploring its potential for long-term cardiac function in the 3-D co-culture model. Chitosan is a natural polysaccharide biomaterial that is biocompatible, biodegradable, non-toxic, and cost effective. Electrospun chitosan was utilized to provide structural scaffolding characterized by scale and architectural resemblance to the extracellular matrix (ECM) in vivo. The chitosan fibers were coated with fibronectin via adsorption in order to enhance cellular adhesion to the fibers and migration into the interfibrous milieu. Ventricular cardiomyocytes were harvested from neonatal rats and studied in various culture conditions (i.e., mono- and co-cultures) for their viability and function. Cellular morphology and functionality were examined using immunofluorescent staining for alpha-sarcomeric actin (SM-actin) and gap junction protein, Connexin-43 (Cx43). Scanning electron microscopy (SEM) and light microscopy were used to investigate cellular morphology, spatial organization, and contractions. Calcium indicator was used to monitor calcium ion flux of beating cardiomyocytes. The results demonstrate that the chitosan nanofibers retained their cylindrical morphology in long-term cell cultures and exhibited good cellular attachment and spreading in the presence of adhesion molecule, fibronectin. Cardiomyocyte mono-cultures resulted in loss of cardiomyocyte polarity and islands of non-coherent contractions. However

JST Thesaurus Headwords and Synonyms: Aeromonas salmonicida [MeCab user dictionary for science technology term[Archive

Lifescience Database Archive (English)

Full Text Available MeCab user dictionary for science technology term Aeromonas salmonicida 名詞 一般 * * *... * Ａｅｒｏｍｏｎａｓ　ｓａｌｍｏｎｉｃｉｄａ Ａｅｒｏｍｏｎａｓ　ｓａｌｍｏｎｉｃｉｄａ エイイーアールオーエムオーエヌエイエス　エスエイエルエムオーエヌアイシーアイディーエイ Thesaurus2015 200906081596351600 C LS07 UNKNOWN_2 Aeromonas salmonicida

Biorelevant media resistant co-culture model mimicking permeability of human intestine.

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Antoine, Delphine; Pellequer, Yann; Tempesta, Camille; Lorscheidt, Stefan; Kettel, Bernadette; Tamaddon, Lana; Jannin, Vincent; Demarne, Frédéric; Lamprecht, Alf; Béduneau, Arnaud

2015-03-15

Cell culture models are currently used to predict absorption pattern of new compounds and formulations in the human gastro-intestinal tract (GIT). One major drawback is the lack of relevant apical incubation fluids allowing mimicking luminal conditions in the GIT. Here, we suggest a culture model compatible with biorelevant media, namely Fasted State Simulated Intestinal Fluid (FaSSIF) and Fed State Simulated Intestinal Fluid (FeSSIF). Co-culture was set up from Caco-2 and mucus-secreting HT29-MTX cells using an original seeding procedure. Viability and cytotoxicity assays were performed following incubation of FeSSIF and FaSSIF with co-culture. Influence of biorelevant fluids on paracellular permeability or transporter proteins were also evaluated. Results were compared with Caco-2 and HT29-MTX monocultures. While Caco-2 viability was strongly affected with FeSSIF, no toxic effect was detected for the co-cultures in terms of viability and lactate dehydrogenase release. The addition of FeSSIF to the basolateral compartment of the co-culture induced cytotoxic effects which suggested the apical mucus barrier being cell protective. In contrast to FeSSIF, FaSSIF induced a slight increase of the paracellular transport and both tested media inhibited partially the P-gp-mediated efflux in the co-culture. Additionally, the absorptive transport of propranolol hydrochloride, a lipophilic β-blocker, was strongly affected by biorelevant fluids. This study demonstrated the compatibility of the Caco-2/HT29-MTX model with some of the current biorelevant media. Combining biorelevant intestinal fluids with features such as mucus secretion, adjustable paracellular and P-gp mediated transports, is a step forward to more realistic in-vitro models of the human intestine. Copyright © 2015. Published by Elsevier B.V.

Neural differentiation of adipose-derived stem cells by indirect co-culture with Schwann cells

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Li Xiaojie

2009-01-01

Full Text Available To investigate whether adipose-derived stem cells (ADSCs could be subject to neural differentiation induced only by Schwann cell (SC factors, we co-cultured ADSCs and SCs in transwell culture dishes. Immunoassaying, Western blot analysis, and RT-PCR were performed (1, 3, 7, 14 d and the co-cultured ADSCs showed gene and protein expression of S-100, Nestin, and GFAP. Further, qRT-PCR disclosed relative quantitative differences in the above three gene expressions. We think ADSCs can undergo induced neural differentiation by being co-cultured with SCs, and such differentia­tions begin 1 day after co-culture, become apparent after 7 days, and thereafter remain stable till the 14th day.

Prolactin family of the guinea pig, Cavia porcellus.

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Alam, S M Khorshed; Konno, Toshihiro; Rumi, M A Karim; Dong, Yafeng; Weiner, Carl P; Soares, Michael J

2010-08-01

Prolactin (PRL) is a multifunctional hormone with prominent roles in regulating growth and reproduction. The guinea pig (Cavia porcellus) has been extensively used in endocrine and reproduction research. Thus far, the PRL cDNA and protein have not been isolated from the guinea pig. In the present study, we used information derived from the public guinea pig genome database as a tool for identifying guinea pig PRL and PRL-related proteins. Guinea pig PRL exhibits prominent nucleotide and amino acid sequence differences when compared with PRLs of other eutherian mammals. In contrast, guinea pig GH is highly conserved. Expression of PRL and GH in the guinea pig is prominent in the anterior pituitary, similar to known expression patterns of PRL and GH for other species. Two additional guinea pig cDNAs were identified and termed PRL-related proteins (PRLRP1, PRLRP2). They exhibited a more distant relationship to PRL and their expression was restricted to the placenta. Recombinant guinea pig PRL protein was generated and shown to be biologically active in the PRL-responsive Nb2 lymphoma cell bioassay. In contrast, recombinant guinea pig PRLRP1 protein did not exhibit PRL-like bioactivity. In summary, we have developed a new set of research tools for investigating the biology of the PRL family in an important animal model, the guinea pig.

Design of biomimetic cellular scaffolds for co-culture system and their application

Science.gov (United States)

Kook, Yun-Min; Jeong, Yoon; Lee, Kangwon; Koh, Won-Gun

2017-01-01

The extracellular matrix of most natural tissues comprises various types of cells, including fibroblasts, stem cells, and endothelial cells, which communicate with each other directly or indirectly to regulate matrix production and cell functionality. To engineer multicellular interactions in vitro, co-culture systems have achieved tremendous success achieving a more realistic microenvironment of in vivo metabolism than monoculture system in the past several decades. Recently, the fields of tissue engineering and regenerative medicine have primarily focused on three-dimensional co-culture systems using cellular scaffolds, because of their physical and biological relevance to the extracellular matrix of actual tissues. This review discusses several materials and methods to create co-culture systems, including hydrogels, electrospun fibers, microfluidic devices, and patterning for biomimetic co-culture system and their applications for specific tissue regeneration. Consequently, we believe that culture systems with appropriate physical and biochemical properties should be developed, and direct or indirect cell–cell interactions in the remodeled tissue must be considered to obtain an optimal tissue-specific microenvironment. PMID:29081966

Presence of Vibrios and Aeromonas strains and total psychrotrophs in hexagonal spotted grouper and Spanish Mackerel Fish

International Nuclear Information System (INIS)

Al-Dagal, M.

2002-01-01

This work was conducted on different parts of hexagonal-spotted grouper and Spanish mackerel fish from 10 major fishery shops in Riyadh city to assess the presence of vibrios and aeromonas using the API20E and Biolog system. Also, pH and total psychotrophs were assessed as two indexes of quality. Similar data were obtained from both identification systems except for Aeromonas media-like and Vibrio anguillarum which are not included in the API20E database. Aeromonas hydrophila, A. media-like, V. alginolyticus, V.anguillarum, V. damsel and V. fluvialis were recovered from fish samples A. medial-like and V. damsel most predominant. Unexpectedly, V. cholera and V. parahemolyticus were not recovered. The numbers of skin samples of grouper having more than 7 log CFU/gm were much higher than that of mackerel skin samples. Similar psychrotrophic counts were noticed in gut samples of the two fish types, indicating similarly in growth habitat. Most of the grouper fish samples had a pH in the range of >6.40-7.02, whereas 95% of mackerel samples had a pH at 6.40 or below. (author)

Occurrence and Variety of β-Lactamase Genes among Aeromonas spp. Isolated from Urban Wastewater Treatment Plant

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Marta Piotrowska

2017-05-01

Full Text Available Members of the genus Aeromonas that commonly occur in various aquatic ecosystems are taken into account as vectors spreading antibiotic resistance genes (ARGs in the environment. In our study strains of Aeromonas spp. (n = 104 not susceptible to ampicillin were isolated from municipal sewage of different levels of purification – raw sewage, activated sludge and treated wastewater. The crucial step of the study was the identification of β-lactamase resistance genes. The identified genes encode β-lactamases from 14 families – blaTEM, blaOXA, blaSHV, blaCTX-M, blaMOX, blaACC, blaFOX, blaGES, blaPER, blaV EB, blaKPC, cphA, imiH, and cepH. There were no significant differences in number of identified ARGs between isolation points. BlaOXA, blaFOX variants and, characteristic for Aeromonas genus, metallo-β-lactamase cphA-related genes were the most commonly identified types of β-lactam resistance determinants. Moreover, we found four extended-spectrum β-lactamases (blaSHV -11, blaCTX-M-27, blaCTX-M-98, and blaPER-4 – and seven AmpC (blaACC, blaFOX-2-like, blaFOX-3, blaFOX-4-like, blaFOX-9, blaFOX-10-like, and blaFOX-13-like types and variants of genes that had never been found among Aeromonas spp. before. Five of the β-lactamases families (blaTEM, blaOXA, blaFOX, blaV EB, and cphA were identified in all three isolation sites, which supports the hypothesis that wastewater treatment plants (WWTPs are hot spots of ARGs dissemination. The obtained ARGs sequences share high identity with previously described β-lactamases, but new variants of those genes have to be considered as well. Characterization of antibiotic susceptibility was performed using disk the diffusion method with 12 different antibiotics according to CLSI guidelines. Over 60% of the strains are unsusceptible to cefepime and chloramphenicol and the majority of the strains have a multidrug resistance phenotype (68%. Finally, analysis of plasmid profiles among the resistant strains

An experimental approach towards the development of an in vitro cortical-thalamic co-culture model.

Science.gov (United States)

Kanagasabapathi, Thirukumaran T; Massobrio, Paolo; Tedesco, Mariateresa; Martinoia, Sergio; Wadman, Wytse J; Decré, Michel M J

2011-01-01

In this paper, we propose an experimental approach to develop an in vitro dissociated cortical-thalamic co-culture model using a dual compartment neurofluidic device. The device has two compartments separated by 10 μm wide and 3 μm high microchannels. The microchannels provide a physical isolation of neurons allowing only neurites to grow between the compartments. Long-term viable co-culture was maintained in the compartmented device, neurite growth through the microchannels was verified using immunofluorescence staining, and electrophysiological recordings from the co-culture system was investigated. Preliminary analysis of spontaneous activities from the co-culture shows a distinctively different firing pattern associated with cultures of individual cell types and further analysis is proposed for a deeper understanding of the dynamics involved in the network connectivity in such a co-culture system.

Decolorization of synthetic brilliant green carpet industry dye through fungal co-culture technology.

Science.gov (United States)

Kumari, Simpal; Naraian, Ram

2016-09-15

Aim of the present study was to evaluate the efficiency of fungal co-culture for the decolorization of synthetic brilliant green carpet industry dye. For this purpose two lignocellulolytic fungi Pleurotus florida (PF) and Rhizoctonia solani (RS) were employed. The study includes determination of enzyme profiles (laccase and peroxidase), dye decolorization efficiency of co-culture and crude enzyme extracts. Both fungi produced laccase and Mn peroxidase and successfully decolorized solutions of different concentrations (2.0, 4.0, 6.0, & 8.0(w/v) of dye. The co-culture resulted highest 98.54% dye decolorization at 2% (w/v) of dye as compared to monocultures (82.12% with PF and 68.89% with RS) during 12 days of submerged fermentation. The lower levels of dyes were rapidly decolorized, while higher levels in slow order as 87.67% decolorization of 8% dye. The promising achievement of the study was remarkable decolorizing efficiency of co-culture over monocultures. The direct treatment of the mono and co-culture enzyme extracts to dye also influenced remarkable. The highest enzymatic decolorization was through combined (PF and RS) extracts, while lesser by monoculture extracts. Based on the observations and potentiality of co-culture technology; further it can be exploited for the bioremediation of areas contaminated with hazardous environmental pollutants including textile and other industry effluents. Copyright © 2016 Elsevier Ltd. All rights reserved.

Purinergic modulation of adult guinea pig cardiomyocytes in long term cultures and co-cultures with extracardiac or intrinsic cardiac neurones.

Science.gov (United States)

Horackova, M; Huang, M H; Armour, J A

1994-05-01

To determine the capacity of ATP to modify cardiomyocytes directly or indirectly via peripheral autonomic neurones, the effects of various purinergic agents were studied on long term cultures of adult guinea pig ventricular myocytes and their co-cultures with extracardiac (stellate ganglion) or intrinsic cardiac neurones. Ventricular myocytes and cardiac neurones were enzymatically dissociated and plated together or alone (myocytes only). Myocyte cultures were used for experiments after three to six weeks. The electrical and contractile properties of cultured myocytes and myocyte-neuronal networks were investigated. The spontaneous beating frequency of ventricular myocytes co-cultured with stellate ganglion neurones increased by approximately 140% (p under control conditions, but when beta adrenergic receptors of tetrodotoxin sensitive neural responses were blocked, ATP induced greater augmentation (> 100%). In contrast, ATP induced much smaller effects in non-innervated myocyte cultures (approximately 26%, p UTP > MSATP > beta gamma ATP > alpha beta ATP. Adenosine (10(-4) M) attenuated the beating frequency of myocytes in both types of co-culture, while not significantly affecting non-innervated myocyte cultures. The experimental model used in this study showed that extrinsic and intrinsic cardiac neurones which possess P2 receptors can greatly enhance cardiac myocyte contractile rate when activated by ATP. Since adenosine reduced contractile rate in both types of co-cultures while not affecting non-innervated myocytes, it is concluded that some of these neurones possess P1 receptors.

Three-dimensional co-culture facilitates the differentiation of embryonic stem cells into mature cardiomyocytes.

Science.gov (United States)

Ou, Dong-Bo; He, Yong; Chen, Rui; Teng, Ji-Wei; Wang, Hong-Tao; Zeng, Di; Liu, Xiong-Tao; Ding, Lu; Huang, Jin-Yan; Zheng, Qiang-Sun

2011-12-01

The cardiomyocyte (CM) differentiation of embryonic stem cells (ESCs) is routinely cultured as two-dimensional (2D) monolayer, which doesn't mimic in vivo physiological environment and may lead to low differentiated level of ESCs. Here, we develop a novel strategy that enhances CM differentiation of ESCs in collagen matrix three-dimensional (3D) culture combined with indirect cardiac fibroblasts co-culture. ESCs were cultured in hanging drops to form embryoid bodies (EBs) and then applied on collagen matrix. The EBs were indirectly co-cultured with cardiac fibroblasts by the hanging cell culture inserts (PET 1 µm). The molecular expressions and ultrastructural characteristics of ESC-derived CMs (ESCMs) were analyzed by real time RT-PCR, immunocytochemistry, and Transmission Electron Microscopy (TEM). We found that the percentage of beating EBs with cardiac fibroblasts co-culture was significantly higher than that without co-culture after differentiation period of 8 days. Type I collagen used as 3D substrates enhanced the late-stage CM differentiation of ESCs and had effect on ultrastructural mature of ESCMs in late-stage development. The combined effects of 3D and co-culture that mimic in vivo physiological environment further improved the efficiency of CM differentiation from ESCs, resulting in fiber-like structures of cardiac cells with organized sarcomeric structure in ESCMs. This novel 3D co-culture system emphasizes the fact that the ESC differentiation is actively responding to cues from their environment and those cues can drive phenotypic control, which provides a useful in vitro model to investigate CM differentiation of stem cells. Copyright © 2011 Wiley Periodicals, Inc.

A yeast co-culture-based biosensor for determination of waste water contamination levels.

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Yudina, N Yu; Arlyapov, V A; Chepurnova, M A; Alferov, S V; Reshetilov, A N

2015-10-01

Artificial microbial co-cultures were formed to develop the receptor element of a biosensor for assessment of biological oxygen demand (BOD). The co-cultures possessed broad substrate specificities and enabled assays of water and fermentation products within a broad BOD range (2.4-80 mg/dm(3)) with a high correlation to the standard method (R = 0.9988). The use of the co-cultures of the yeasts Pichia angusta, Arxula adeninivorans and Debaryomyces hansenii immobilized in N-vinylpyrrolidone-modified poly(vinyl alcohol) enabled developing a BOD biosensor possessing the characteristics not inferior to those in the known biosensors. The results are indicative of a potential of using these co-cultures as the receptor element base in prototype models of instruments for broad application. Copyright © 2015 Elsevier Inc. All rights reserved.

Role of pH in oxidase variability of Aeromonas hydrophila.

OpenAIRE

Hunt, L K; Overman, T L; Otero, R B

1981-01-01

Some strains of Aeromonas hydrophila may be oxidase negative or only weakly oxidase positive by the Kovacs method taken from the surface of a differential medium, such as MacConkey agar. Six strains of A. hydrophila, two oxidase variable, one oxidase constant, and three weakly oxidase positive on MacConkey agar, were studied to determine the cause of oxidase variability. The bacteriostatic dyes in MacConkey agar were considered possible inhibitors of the oxidase reaction. The concentration of...

Use of bacterial co-cultures for the efficient production of chemicals.

Science.gov (United States)

Jones, J Andrew; Wang, Xin

2017-12-02

The microbial production of chemicals has traditionally relied on a single engineered microbe to enable the complete bioconversion of substrate to final product. Recently, a growing fraction of research has transitioned towards employing a modular co-culture engineering strategy using multiple microbes growing together to facilitate a divide-and-conquer approach for chemical biosynthesis. Here, we review key success stories that leverage the unique advantages of co-culture engineering, while also addressing the critical concerns that will limit the wide-spread implementation of this technology. Future studies that address the need to monitor and control the population dynamics of each strain module, while maintaining robust flux routes towards a wide range of desired products will lead the efforts to realize the true potential of co-culture engineering. Copyright © 2017 Elsevier Ltd. All rights reserved.

Social preferences of developing guinea pigs (Cavia porcellus) from the preweaning to the periadolescent periods.

Science.gov (United States)

Hennessy, Michael B; Young, Travis L; O'Leary, Shonagh K; Maken, Deborah S

2003-12-01

Preference tests in a novel environment (Experiment 1) and unobtrusive observations in a specialized living environment (Experiment 2) examined the attractiveness of various classes of conspecifics for maturing guinea pigs (Cavia porcellus). It was found that (a). the young continued to remain near the mother well beyond weaning; (b). there was increased time spent with unrelated adult females, but not males, after weaning; (c). male and female offspring behaved similarly; and (d). littermates spent considerable time with each other. These results provide no evidence that guinea pigs approaching sexual maturity begin to associate preferentially with novel animals or potential breeding partners. Choices were largely predictable from earlier findings of the ability of various classes of conspecifics to reduce hypothalamic-pituitary-adrenal activity of the young.

Resistance of Salmonds Against Aeromonas Salmonicida

DEFF Research Database (Denmark)

Holten-Andersen, Lars; Dalsgaard, Inger; Buchmann, Kurt

2011-01-01

Furunculosis, caused by Aeromonas salmonicida, continues to be a health problem for the growing salmonid aquaculture. Despite effective vaccination programs regular outbreaks occur at Danish trout farms calling for repeated antibiotic treatment. We hypothesized that a difference in natural...... over a 30-day period post challenge and the results were summarized in mortality curves. A. salmonicida was recovered from mortalities during the entire test period. At day 30 the survival was 6.2 % and 34.0 % for rainbow trout and Baltic salmon, respectively. Significant differences in susceptibility...... to A. salmonicida were demonstrated between the two salmonids and hazard ratio estimation between rainbow trout and Baltic salmon showed a 3.36 higher risk of dying from the infection in the former. The finding that Baltic salmon carries a high level of natural resistance to furunculosis might raise...

Commentary on “Co-creating Stakeholder and Brand Identities: A Cross-cultural Consumer Perspective”

DEFF Research Database (Denmark)

Csaba, Fabian

2017-01-01

This commentary raises awareness for the relevance of other cultural dimensions- besides individualism and collectivism - and alternative approaches to cross-cultural research for exploring cultural variations in stakeholders' co-construction of brand identity and their own identities. The author...... influence of various cultures on each other can provide additional, relevant insights into reciprocal identity co-construction processes between brands and stakeholders....

Reação em cadeia da polimerase para detecção de Clostridium chauvoei em tecidos de Cavia porcellus PCR detection of Clostridium chauvoei in tissues of Cavia porcellus

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Ronnie Antunes de Assis

2005-12-01

Full Text Available O objetivo deste trabalho foi padronizar uma técnica de reação em cadeia da polimerase (PCR, para detecção de Clostridium chauvoei, em tecidos fixados em formol e incluídos em parafina de cobaias (Cavia porcellus experimentalmente infectadas com esse microrganismo. Os animais foram sacrificados, e amostras do músculo da área de inoculação (MAI, fígado, miocárdio e baço foram disponibilizadas para a técnica de PCR. O clostrídio foi detectado em todas as secções do MAI, fígado e miocárdio, mas não foi observado em secções do baço. Reações cruzadas não foram observadas a partir de secções do MAI dos animais com inoculação de outras espécies de clostrídios, bem como nenhuma amplificação foi observada a partir de secções do MAI dos animais controle. Esses resultados mostram que a técnica de PCR desenvolvida neste estudo, pode ser usada para detecção de Clostridium chauvoei em tecidos fixados em formol e incluídos em parafina.The objective of this work was the standardization of a polymerase chain reaction (PCR for detection of Clostridium chauvoei in formalin-fixed, paraffin-embedded tissues of guinea-pigs (Cavia porcellus infected experimentally with this microorganism. The animals were sacrificed, and samples of muscle from inoculation area (MIA, liver, myocardium and spleen were available for PCR technique. Clostridium chauvoei was detected in all sections of the MIA, liver and myocardium, and no product was observed in sections of the spleen. Cross-reactions were not observed in sections of MIA of the animals inoculated with other clostridia, as well as no amplification was observed in sections of MIA of control animals. These results show that the PCR technique developed in this study may be useful for detection of C. chauvoei in formalin-fixed, paraffin-embedded tissues.

In vitro co-cultures of human gut bacterial species as predicted from co-occurrence network analysis

DEFF Research Database (Denmark)

Das, Promi; Ji, Boyang; Kovatcheva-Datchary, Petia

2018-01-01

Network analysis of large metagenomic datasets generated by current sequencing technologies can reveal significant co-occurrence patterns between microbial species of a biological community. These patterns can be analyzed in terms of pairwise combinations between all species comprising a community...... thetaiotaomicron, as well as Faecalibacterium prausnitzii and Roseburia inulinivorans as model organisms for our study. We then delineate the outcome of the co-cultures when equal distributions of resources were provided. The growth behavior of the co-culture was found to be dependent on the types of microbial...... species present, their specific metabolic activities, and resulting changes in the culture environment. Through this reductionist approach and using novel in vitro combinations of microbial species under anaerobic conditions, the results of this work will aid in the understanding and design of synthetic...

[Biological characteristics of mesenchymal stem cell and hematopoietic stem cell in the co-culture system].

Science.gov (United States)

Wei, Wei; Xu, Chao; Ye, Zhi-Yong; Huang, Xiao-Jun; Yuan, Jia-En; Ma, Tian-Bao; Lin, Han-Biao; Chen, Xiu-Qiong

2016-10-25

The aim of the present study was to obtain the qualified hematopoietic stem/progenitor cells (HSC/HPC) and human umbilical cord-mesenchymal stem cells (MSC) in vitro in the co-culture system. Cord blood mononuclear cells were separated from umbilical cord blood by Ficoll lymphocyte separation medium, and then CD34 + HSC was collected by MACS immunomagnetic beads. The selected CD34 + HSC/HPC and MSC were transferred into culture flask. IMDM culture medium with 15% AB-type cord plasma supplemented with interleukin-3 (IL-3), IL-6, thrombopoietin (TPO), stem cell factor (SCF) and FMS-like tyrosine kinase 3 ligand (Flt-3L) factors were used as the co-culture system for the amplification of HSC/HPC and MSC. The cellular growth status and proliferation on day 6 and 10 after co-culture were observed by using inverted microscope. The percentage of positive expression of CD34 in HSC/HPC, as well as the percentages of positive expressions of CD105, CD90, CD73, CD45, CD34 and HLA-DR in the 4 th generation MSC, was tested by flow cytometry. Semisolid colony culture was used to test the HSC/HPC colony forming ability. The osteogenic, chondrogenesis and adipogenic ability of the 4 th generation MSC were assessed. The karyotype analysis of MSC was conducted by colchicines. The results demonstrated that the HSC/HPC of co-culture group showed higher ability of amplification, CFU-GM and higher CD34 + percentage compared with the control group. The co-cultured MSC maintained the ability to differentiate into bone cells, fat cells and chondrocytes. And the karyotype stability of MSC remained normal. These results reveal that the appropriate co-culture system for MSC and HSC is developed, and via this co-culture system we could gain both two kinds of these cells. The MSCs under the co-culture system maintain the biological characteristics. The CFU-GM ability, cell counting and the flow cytometry results of HSC/HPC under the co-culture system are conform to the criterion, showing that

Plesiomonasshigelloides and Aeromonadaceae family pathogens isolated from marine mammals of Southern and Southeastern Brazilian coast Plesiomonasshigelloides e patógenos da família Aeromonadaceae isolados de mamíferos marinhos da costa sul e sudeste do Brasil

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Christiane S. Pereira

2008-12-01

Full Text Available The aquatic environment is the habitat of many microorganisms, including Plesiomonasshigelloides and Aeromonas species which are pathogenic to human and animals. In the present investigation, we evaluated the occurrence of these pathogens from marine mammals beached or accidentally captured by fishing net in southeastern (RJ and southern (RS coastal Brazilian regions. A total of 198 swabs from 27 specimens of marine mammals, including 11 different species, were collected by DEENSP and GEMARS-CECLIMAR/UFRGS Institutes and sent to LRNCEB/IOC/FIOCRUZ. The samples were enriched in Alkaline Peptone Water (APW added with 1% of sodium chloride (NaCl, APW plus 3% NaCl and incubated at 37ºC for 18-24 hours. Following, samples were streaked onto Pseudomonas-Aeromonas Selective Agar Base (GSP Agar and suspected colonies were biochemically characterized. The results revealed 114 strains, including ten Aeromonas species and P.shigelloides. The main pathogens isolated were A.veronii biogroup veronii (19.3%, A. caviae (12.3%, A. hydrophila (9.6% and P.shigelloides (7%. The pathogens were isolated in both coastal and offshore marine mammals. These data point the importance of epidemiological surveillance and microbiological monitoring and reinforce the need to implement environmental protection programs, especially related to endangered cetacean species.O ambiente aquático é o habitat de vários microrganismos, incluindo Plesiomonasshigelloides e espécies de Aeromonas, os quais são patogênicos para o homem e os animais. Na presente investigação, foi avaliada a ocorrência destes patógenos a partir de swabs coletados de mamíferos marinhos encalhados ou capturados acidentalmente em redes de pesca nas regiões costeiras do sudeste (RJ e sul (RS do Brasil. O total de 198 swabs de 27 espécimes de mamíferos marinhos, incluindo 11 espécies distintas, foi coletado por profissionais dos institutos DEENSP, GEMARS-CECLIMAR/UFRGS e enviado ao LRNCEB

Co-culture of 3D tumor spheroids with fibroblasts as a model for epithelial–mesenchymal transition in vitro

International Nuclear Information System (INIS)

Kim, Sun-Ah; Lee, Eun Kyung; Kuh, Hyo-Jeong

2015-01-01

Epithelial–mesenchymal transition (EMT) acts as a facilitator of metastatic dissemination in the invasive margin of malignant tumors where active tumor–stromal crosstalks take place. Co-cultures of cancer cells with cancer-associated fibroblasts (CAFs) are often used as in vitro models of EMT. We established a tumor–fibroblast proximity co-culture using HT-29 tumor spheroids (TSs) with CCD-18co fibroblasts. When co-cultured with TSs, CCD-18co appeared activated, and proliferative activity as well as cell migration increased. Expression of fibronectin increased whereas laminin and type I collagen decreased in TSs co-cultured with fibroblasts compared to TSs alone, closely resembling the margin of in vivo xenograft tissue. Active TGFβ1 in culture media significantly increased in TS co-cultures but not in 2D co-cultures of cancer cells–fibroblasts, indicating that 3D context-associated factors from TSs may be crucial to crosstalks between cancer cells and fibroblasts. We also observed in TSs co-cultured with fibroblasts increased expression of α-SMA, EGFR and CTGF; reduced expression of membranous β-catenin and E-cadherin, together suggesting an EMT-like changes similar to a marginal region of xenograft tissue in vivo. Overall, our in vitro TS–fibroblast proximity co-culture mimics the EMT-state of the invasive margin of in vivo tumors in early metastasis. - Highlights: • An adjacent co-culture of tumor spheroids and fibroblasts is presented as EMT model. • Activation of fibroblasts and increased cell migration were shown in co-culture. • Expression of EMT-related factors in co-culture was similar to that in tumor tissue. • Crosstalk between spheroids and fibroblasts was demonstrated by secretome analysis

Co-culture of 3D tumor spheroids with fibroblasts as a model for epithelial–mesenchymal transition in vitro

Energy Technology Data Exchange (ETDEWEB)

Kim, Sun-Ah, E-mail: j.sarah.k@gmail.com [Department of Biomedicine & Health Sciences, College of Medicine, The Catholic University of Korea, Seoul 137-701 (Korea, Republic of); Lee, Eun Kyung, E-mail: leeek@catholic.ac.kr [Department of Biochemistry, College of Medicine, The Catholic University of Korea, Seoul 137-701 (Korea, Republic of); Cancer Evolution Research Center, College of Medicine, The Catholic University of Korea, Seoul 137-701 (Korea, Republic of); Kuh, Hyo-Jeong, E-mail: hkuh@catholic.ac.kr [Department of Biomedicine & Health Sciences, College of Medicine, The Catholic University of Korea, Seoul 137-701 (Korea, Republic of); Cancer Evolution Research Center, College of Medicine, The Catholic University of Korea, Seoul 137-701 (Korea, Republic of)

2015-07-15

Epithelial–mesenchymal transition (EMT) acts as a facilitator of metastatic dissemination in the invasive margin of malignant tumors where active tumor–stromal crosstalks take place. Co-cultures of cancer cells with cancer-associated fibroblasts (CAFs) are often used as in vitro models of EMT. We established a tumor–fibroblast proximity co-culture using HT-29 tumor spheroids (TSs) with CCD-18co fibroblasts. When co-cultured with TSs, CCD-18co appeared activated, and proliferative activity as well as cell migration increased. Expression of fibronectin increased whereas laminin and type I collagen decreased in TSs co-cultured with fibroblasts compared to TSs alone, closely resembling the margin of in vivo xenograft tissue. Active TGFβ1 in culture media significantly increased in TS co-cultures but not in 2D co-cultures of cancer cells–fibroblasts, indicating that 3D context-associated factors from TSs may be crucial to crosstalks between cancer cells and fibroblasts. We also observed in TSs co-cultured with fibroblasts increased expression of α-SMA, EGFR and CTGF; reduced expression of membranous β-catenin and E-cadherin, together suggesting an EMT-like changes similar to a marginal region of xenograft tissue in vivo. Overall, our in vitro TS–fibroblast proximity co-culture mimics the EMT-state of the invasive margin of in vivo tumors in early metastasis. - Highlights: • An adjacent co-culture of tumor spheroids and fibroblasts is presented as EMT model. • Activation of fibroblasts and increased cell migration were shown in co-culture. • Expression of EMT-related factors in co-culture was similar to that in tumor tissue. • Crosstalk between spheroids and fibroblasts was demonstrated by secretome analysis.

Antibiotic resistance profiling and phenotyping of Aeromonas species isolated from aquatic sources

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Olumide A. Odeyemi

2017-01-01

Full Text Available This study aimed to investigate antibiotics resistance pattern and phenotyping of Aeromonas species isolated from different aquatic sources in Melaka, Malaysia. A total of 53 Aeromonas species were isolated from the following sources: sediment (n = 13, bivalve (n = 10, sea cucumber (n = 16 and sea water (n = 14 and resistance to 12 antibiotics – Tetracycline (30 μg, Kanamycin (30 μg, Oxytetracycline (30 μg, Ampicillin (10 μg, Streptomycin (10 μg, Gentamicin (10 μg, Sulphamethoxazole (25 μg, Nalixidic acid (30 μg, Trimethoprim (1.25 μg, Novobiocin (5 μg, Penicilin (10 μg and Chloramphenicol (10 μg was tested. The results obtained from this study reveal multi drug resistance pattern among the isolates. All the isolates were completely resistant to Ampicillin, Novobiocin, Sulphamethoxazole and Trimethoprim, respectively but susceptible to Tetracycline (100%, Kanamycin (5.7%, Gentamicin (5.7% and Oxytetracycline (24.5%. Antibiotics phenotyping of the bacteria revealed 21 different phenotypes among the isolates.

Co-culture with infrapatellar fat pad differentially stimulates proteoglycan synthesis and accumulation in cartilage and meniscus tissues.

Science.gov (United States)

Nishimuta, James F; Bendernagel, Monica F; Levenston, Marc E

2017-09-01

Although osteoarthritis is widely viewed as a disease of the whole joint, relatively few studies have focused on interactions among joint tissues in joint homeostasis and degeneration. In particular, few studies have examined the effects of the infrapatellar fat pad (IFP) on cartilaginous tissues. The aim of this study was to test the hypothesis that co-culture with healthy IFP would induce degradation of cartilage and meniscus tissues. Bovine articular cartilage, meniscus, and IFP were cultured isolated or as cartilage-fat or meniscus-fat co-cultures for up to 14 days. Conditioned media were assayed for sulfated glycosaminoglycan (sGAG) content, nitrite content, and matrix metalloproteinase (MMP) activity, and explants were assayed for sGAG and DNA contents. Co-cultures exhibited increased cumulative sGAG release and sGAG release rates for both cartilage and meniscus, and the cartilage (but not meniscus) exhibited a substantial synergistic effect of co-culture (sGAG release in co-culture was significantly greater than the summed release from isolated cartilage and fat). Fat co-culture did not significantly alter the sGAG content of either cartilage or meniscus explants, indicating that IFP co-culture stimulated net sGAG production by cartilage. Nitrite release was increased relative to isolated tissue controls in co-cultured meniscus, but not the cartilage, with no synergistic effect of co-culture. Interestingly, MMP-2 production was decreased by co-culture for both cartilage and meniscus. This study demonstrates that healthy IFP may modulate joint homeostasis by stimulating sGAG production in cartilage. Counter to our hypothesis, healthy IFP did not promote degradation of either cartilage or meniscus tissues.

A defined co-culture of Geobacter sulfurreducens and Escherichia coli in a membrane-less microbial fuel cell.

Science.gov (United States)

Bourdakos, Nicholas; Marsili, Enrico; Mahadevan, Radhakrishnan

2014-04-01

Wastewater-fed microbial fuel cells (MFCs) are a promising technology to treat low-organic carbon wastewater and recover part of the chemical energy in wastewater as electrical power. However, the interactions between electrochemically active and fermentative microorganisms cannot be easily studied in wastewater-fed MFCs because of their complex microbial communities. Defined co-culture MFCs provide a detailed understanding of such interactions. In this study, we characterize the extracellular metabolites in laboratory-scale membrane-less MFCs inoculated with Geobacter sulfurreducens and Escherichia coli co-culture and compare them with pure culture MFCs. G. sulfurreducens MFCs are sparged to maintain anaerobic conditions, while co-culture MFCs rely on E. coli for oxygen removal. G. sulfurreducens MFCs have a power output of 128 mW m(-2) , compared to 63 mW m(-2) from the co-culture MFCs. Analysis of metabolites shows that succinate production in co-culture MFCs decreases current production by G. sulfurreducens and that the removal of succinate is responsible for the increased current density in the late co-culture MFCs. Interestingly, pH adjustment is not required for co-culture MFCs but a base addition is necessary for E. coli MFCs and cultures in vials. Our results show that defined co-culture MFCs provide clear insights into metabolic interactions among bacteria while maintaining a low operational complexity. © 2013 Wiley Periodicals, Inc.

El pH de la carne de cobayo (Cavia porcellus procedente del manejo deficiente del bienestar animal durante el sacrificio en la sierra central del Perú.

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LUCAS JR

2017-09-01

Full Text Available En la región andina del Perú es común observar el sacrificio de los cobayos sin aturdimiento ni descanso ante mortem. El objetivo del presente estudio fue evaluar el pH de la carne de cobayo (Cavia porcellus resultante de este manejo. Se evaluó el pH de 60 canales a las 0, 1, 6, 12, y 24 h posteriores al sacrificio sin descanso ante mortem ni aturdimiento. El pH fue de 6.79 ± 0.3, 6.74 ± 0.34, 6.59 ± 0.28, 6.45 ± 0.35 y 5.96 ± 0.11, a la 0, 1, 6, 12 y 24 horas post sacrificio respectivamente. La ecuación que representa la evolución del pH fue Y=6.7765-0.0231X-0.0005X2 (R2=0.996. SUMMARY. The pH of guinea pig (Cavia porcellus meat from the welfare-deficient slaughtering in the Peruvian central Andes. In Andean region of Peru is common to slaughter Guinea pig without lairage and stunning. The aim of this study was to evaluate the pH of meat of Guinea pig (Cavia porcellus from the unsuitable welfare practices during slaughtering in the Peruvian central Andes. The pH of 60 guinea pig carcasses were analyzed at 0, 1, 6, 12 and 24 hours after the end of the slaughter process. The pH was 6.79 ± 0.3, 6.74 ± 0.34, 6.59 ± 0.28, 6.45 ± 0.35 and 5.96 ± 0.11 for 0, 1, 6, 12 y 24 h post slaughtering. The equation that graphs pH evolution was Y=6.7765-0.0231X-0.0005X2 (R2=0.996.

Invasion fitness for gene-culture co-evolution in family-structured populations and an application to cumulative culture under vertical transmission.

Science.gov (United States)

Mullon, Charles; Lehmann, Laurent

2017-08-01

Human evolution depends on the co-evolution between genetically determined behaviors and socially transmitted information. Although vertical transmission of cultural information from parent to offspring is common in hominins, its effects on cumulative cultural evolution are not fully understood. Here, we investigate gene-culture co-evolution in a family-structured population by studying the invasion fitness of a mutant allele that influences a deterministic level of cultural information (e.g., amount of knowledge or skill) to which diploid carriers of the mutant are exposed in subsequent generations. We show that the selection gradient on such a mutant, and the concomitant level of cultural information it generates, can be evaluated analytically under the assumption that the cultural dynamic has a single attractor point, thereby making gene-culture co-evolution in family-structured populations with multigenerational effects mathematically tractable. We apply our result to study how genetically determined phenotypes of individual and social learning co-evolve with the level of adaptive information they generate under vertical transmission. We find that vertical transmission increases adaptive information due to kin selection effects, but when information is transmitted as efficiently between family members as between unrelated individuals, this increase is moderate in diploids. By contrast, we show that the way resource allocation into learning trades off with allocation into reproduction (the "learning-reproduction trade-off") significantly influences levels of adaptive information. We also show that vertical transmission prevents evolutionary branching and may therefore play a qualitative role in gene-culture co-evolutionary dynamics. More generally, our analysis of selection suggests that vertical transmission can significantly increase levels of adaptive information under the biologically plausible condition that information transmission between relatives is

Human pericyte-endothelial cell interactions in co-culture models mimicking the diabetic retinal microvascular environment.

Science.gov (United States)

Tarallo, Sonia; Beltramo, Elena; Berrone, Elena; Porta, Massimo

2012-12-01

Pericytes regulate vascular tone, perfusion pressure and endothelial cell (EC) proliferation in capillaries. Thiamine and benfotiamine counteract high glucose-induced damage in vascular cells. We standardized two human retinal pericyte (HRP)/EC co-culture models to mimic the diabetic retinal microvascular environment. We aimed at evaluating the interactions between co-cultured HRP and EC in terms of proliferation/apoptosis and the possible protective role of thiamine and benfotiamine against high glucose-induced damage. EC and HRP were co-cultured in physiological glucose and stable or intermittent high glucose, with or without thiamine/benfotiamine. No-contact model: EC were plated on a porous membrane suspended into the medium and HRP on the bottom of the same well. Cell-to-cell contact model: EC and HRP were plated on the opposite sides of the same membrane. Proliferation (cell counts and DNA synthesis), apoptosis and tubule formation in Matrigel were assessed. In the no-contact model, stable high glucose reduced proliferation of co-cultured EC/HRP and EC alone and increased co-cultured EC/HRP apoptosis. In the contact model, both stable and intermittent high glucose reduced co-cultured EC/HRP proliferation and increased apoptosis. Stable high glucose had no effects on HRP in separate cultures. Both EC and HRP proliferated better when co-cultured. Thiamine and benfotiamine reversed high glucose-induced damage in all cases. HRP are sensitive to soluble factors released by EC when cultured in high glucose conditions, as suggested by conditioned media assays. In the Matrigel models, addition of thiamine and benfotiamine re-established the high glucose-damaged interactions between EC/HRP and stabilized microtubules.

Evaluation of Medicinal Plant Hepatotoxicity in Co-cultures of Hepatocytes and Monocytes

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Bashar Saad

2006-01-01

Full Text Available Non-parenchymal cells might play an important role in the modulation of xenobiotic metabolism in liver and its pharmacological and toxicological consequences. Therefore, the role of cell-to-cell interactions in herbal induced liver toxicity was investigated in monocultures of cells from the human hepatocyte cell line (HepG2 and in co-cultures of cells from the HepG2 cell line and cells from the human monocyte cell line (THP1. Cells were treated with various concentrations (1–500 µg ml−1 of extracts of Pistacia palaestina, Juglans regia and Quercus ithaburensis for 24 h. Extracts from Cleome droserifolia, a known toxic plant, were taken as positive control. In the co-culture system, toxic effects were observed after exposure to extracts of Pistacia palestina and C. droserifolia. These two extracts significantly reduced by cell viability as measured the MTT test and the LDH assay. Whereas in hepatocyte cultures, only extracts of C. droserifolia were found to affect the cell viability. The production levels of albumin from hepatocytes were not affected by treatment with plant extracts in both culture systems. It seems that the observed reduction in cell viability after exposure to extracts of P. palestina in co-cultures but not in monocultures is a result of monocyte-derived factors. The use of liver cell co-cultures is therefore a useful approach to investigate the influence of intercellular communication on xenobiotic metabolism in liver.

Aeromonas hydrophila as cause of hemorrhagic septicemia in a ground-hornbill (Bucorvus abyssinicus).

Science.gov (United States)

Ocholi, R A; Kalejaiye, J O

1990-01-01

An acute hemorrhagic septicemia in a captive ground-hornbill (Bucorvus abyssinicus) is reported. Aeromonas hydrophila was isolated from lungs, liver, kidney, and intestines. Postmortem lesions were characterized by hemorrhages in the internal organs. Microscopically, necrotic lesions were found in the liver, lungs, and kidney. Factors that might have contributed to the clinical disease condition are discussed.

THE PREVALENCE OF Salmonella sp., Listeria sp. AND Aeromonas spp. IN CATFISH (CLARIAS (Clarias gariepinus AND TILAPIA (Tilapia mossambica BY PELLETING METHOD

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Titik Budiati

2013-06-01

Full Text Available The aim of this study was to modify the isolation methods of Aeromonas sp., Salmonella spp., and Listeria sp. in catfish (Clarias gariepinus and tilapia (Tilapia mossambica obtained from wet markets and ponds in Malaysia by pelleting the sample. A total of 108 samples (32 catfish intestines, 32 tilapia intestines, and 44 water samples were obtained from nine wet markets and eight ponds. The modified method was employed by pelleting the samples and followed by either implementing pre-enrichment or without pre-enrichment on the isolation of Salmonella and Listeria spp. The modified method (by pelleting the sample in combination with pre-enrichment was the most efficient for Salmonella and Listeria isolation. The sensitivity of the modified Salmonella isolation method was 0.53 and 0.73 for fish and water samples, respectively. The sensitivity of the modified Listeria method was 1 and 0.92 for fish and water samples, respectively. However, the sensitivity of the method by pelleting the sample was similar to those of non-pelleting the sample on Aeromonas isolation. Five species of Aeromonas spp., seven serovars of Salmonella sp., and four species of Listeria sp. were observed in catfish, tilapia and water samples. Overall, by pelleting the sample offered the beneficial to isolate Aeromonas spp., Salmonella sp. and Listeria spp. in catfish, tilapia and water.

POTENTIAL FOR GREAT EGRETS (ARDEA ALBA) TO TRANSMIT A VIRULENT STRAIN OF AEROMONAS HYDROPHILA AMONG CHANNEL CATFISH (ICTALURUS PUNCTATUS) CULTURE PONDS.

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Jubirt, Madison M; Hanson, Larry A; Hanson-Dorr, Katie C; Ford, Lorelei; Lemmons, Scott; Fioranelli, Paul; Cunningham, Fred L

2015-07-01

Aeromonas hydrophila is a gram-negative, rod-shaped, facultative, anaerobic bacterium that is ubiquitous in freshwater and slightly brackish aquatic environments and infects fish, humans, reptiles, and birds. Recent severe outbreaks of disease in commercial channel catfish (Ictalurus punctatus) aquaculture ponds have been associated with a highly virulent A. hydrophila strain (VAH), which is genetically distinct from less-virulent strains. The epidemiology of this disease has not been determined. Given that A. hydrophila infects birds, we hypothesized that fish-eating birds may serve as a reservoir for VAH and spread the pathogen by flying to uninfected ponds. Great Egrets (Ardea alba) were used in this transmission model because these wading birds frequently prey on farmed catfish. Great Egrets that were fed VAH-infected catfish shed VAH in feces demonstrating their potential to spread VAH.

Chondrogenesis of synovium-derived mesenchymal stem cells in gene-transferred co-culture system.

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Varshney, Rohan R; Zhou, Ruijie; Hao, Jinghua; Yeo, Suan Siong; Chooi, Wai Hon; Fan, Jiabing; Wang, Dong-An

2010-09-01

A co-culture strategy has been developed in this study wherein rabbit synovial mesenchymal stem cells (SMSCs) are co-cultured with growth factor (GF) transfected articular chondrocytes. Toward this end, both SMSCs and early passage rabbit articular chondrocytes that had been adenovirally transduced with transforming growth factor-beta 3 (TGF-beta3) gene were separately encapsulated in alginate beads and co-cultured in the same pool of chondrogenic medium. The chondrocytes act as transfected companion cells (TCCs) providing GF supply to induce chondrogenic differentiation of SMSCs that play the role of therapeutic progenitor cells (TPCs). Against the same TCC based TGF-beta3 release profile, the co-culture was started at different time points (Day 0, Day 10 and Day 20) but made to last for identical periods of exposure (30 days) so that the exposure conditions could be optimized in terms of initiation and duration. Transfection of TCCs prevents the stem cell based TPCs from undergoing the invasive procedure. It also prevents unpredictable complications in the TPCs caused by long-term constitutive over-expression of a GF. The adenovirally transfected TCCs exhibit a transient GF expression which results in a timely termination of GF supply to the TPCs. The TCC-sourced transgenic TGF-beta3 successfully induced chondrogenesis in the TPCs. Real-time PCR results show enhanced expression of cartilage markers and immuno/histochemical staining for Glycosaminoglycans (GAG) and Collagen II also shows abundant extracellular matrix (ECM) production and chondrogenic morphogenesis in the co-cultured TPCs. These results confirm the efficacy of directing stem cell differentiation towards chondrogenesis and cartilage tissue formation by co-culturing them with GF transfected chondrocytes.

Diarrea del viajero asociada a Aeromonas hydrophila Traveller's diarrohea caused by Aeromonas hydrophila

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M. L. Pita Toledo

2001-06-01

Full Text Available Aeromonas hydrophila es una bacteria que se encuentra frecuentemente en aguas dulces y salobres. Desde hace varios años se conoce que determinadas cepas de la especie pueden causar enfermedad en pescados y en anfibios, existiendo cierta controversia sobre su patogeneicidad en humanos. No obstante, puede dar lugar a infección en personas, que la adquieren bien a través de heridas abiertas o bien por ingestión de alimentos o agua contaminados. Sin embargo, en prácticamente la totalidad de las publicaciones se hace referencia a casos aislados más que a brotes producidos por esta bacteria. Generalmente la infección por A. hydrophila da lugar a gastroenteritis en personas sanas, pudiendo generar una septicemia en individuos inmunodeprimidos.En 1998 se produjo una alarma en el sector hotelero por un incremento considerable de los casos de gastroenteritis entre los turistas que acudían durante sus vacaciones a determinadas zonas costeras de la isla de Tenerife.Tras tener conocimiento de la situación, la Dirección General de Salud Pública inició una investigación epidemiológica con el objetivo de conocer la magnitud real del problema detectado e identificar los factores asociados a la misma, para poner en marcha las medidas correctoras y de control necesarias. El análisis inicial permitió determinar que se trataba de un proceso de gran magnitud en su extensión aunque de carácter leve, que afectaba únicamente a los turistas en una zona costera en concreto, compatible por lo tanto con la diarrea del viajero, planteándose un origen probablemente hídrico.Aeromonas hydrophila is a bacteria frequently found in fresh water and saline water. Certain strains of the species have been known to cause disease in fish and amphibians for several years, although some controversy exists about its ability to cause illness in humans. Nevertheless, either through open wounds or contaminated food or water, it can give rise to infection. However, almost

Garlic exerts allelopathic effects on pepper physiology in a hydroponic co-culture system

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Haiyan Ding

2016-05-01

Full Text Available A hydroponic co-culture system was adopted to determine the allelopathic potential of garlic on the growth of pepper plants. Different numbers of garlic plants (0, 2, 4, 8 and 12 were hydroponically co-cultured with two pepper plants to investigate allelopathic effects on the growth attributes and antioxidative defense system of the test pepper plants. The responses of the pepper plants depended on the number of garlic plants included in the co-culture system, indicating an association of pepper growth with the garlic root exudate concentration. When grown at a pepper/garlic ratio of 1:1 or 1:2, the pepper plant height, chlorophyll content, and peroxidase (POD, catalase (CAT and phenylalanine ammonia-lyase (PAL activities were significantly increased after 30 days of co-culture; in contrast, reduction in methane dicarboxylic aldehyde (MDA content was observed. However, when the pepper/garlic ratio was 1:4 or higher, these morphological indices and protective enzyme activities were significantly inhibited, whereas MDA levels in the pepper leaves were significantly increased due to severe membrane lipid peroxidation. The results indicate that although low concentrations of garlic root exudates appear to induce protective enzyme systems and promote pepper growth, high concentrations have deleterious effects. These findings suggest that further investigations should optimize the co-culture pepper/garlic ratio to reduce continuous cropping obstacles in pepper production.

Co-culture of microalgae, cyanobacteria, and macromycetes for exopolysaccharides production: process preliminary optimization and partial characterization.

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Angelis, S; Novak, A C; Sydney, E B; Soccol, V T; Carvalho, J C; Pandey, A; Noseda, M D; Tholozan, J L; Lorquin, J; Soccol, C R

2012-07-01

In this study, the biomass and exopolysaccharides (EPS) production in co-cultures of microalgae/cyanobacteria and macromycetes was evaluated as a technology for producing new polysaccharides for medical and/or industrial application. Based on biomass and EPS productivity of monocultures, two algae and two fungi were selected and cultured in different co-culture arrangements. The hydrosoluble EPS fractions from mono- and co-cultures were characterized by ¹³C NMR spectroscopy and gas chromatography coupled to mass spectrometry and compared. It was found that co-cultures resulted in the production of an EPS different from those produced by monocultures, showing fungal predominance with microalgal/cyanobacterial traces. Co-cultures conditions were screened (temperature, agitation speed, fungal and microalgae inoculation rate, initial pH, illumination rate, and glucose concentration) in order to achieve maximum biomass and EPS production, resulting in an increase of 33 and 61% in exopolysaccharides and biomass productions, respectively (patent pending).

Knockout of the alanine racemase gene in Aeromonas hydrophila HBNUAh01 results in cell wall damage and enhanced membrane permeability.

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Liu, Dong; Zhang, Lu; Xue, Wen; Wang, Yaping; Ju, Jiansong; Zhao, Baohua

2015-07-01

This study focused on the alanine racemase gene (alr-2), which is involved in the synthesis of d-alanine that forms the backbone of the cell wall. A stable alr-2 knockout mutant of Aeromonas hydrophila HBNUAh01 was constructed. When the mutant was supplemented with d-alanine, growth was unaffected; deprivation of d-alanine caused the growth arrest of the starved mutant cells, but not cell lysis. No alanine racemase activity was detected in the culture of the mutant. Additionally, a membrane permeability assay showed increasing damage to the cell wall during d-alanine starvation. No such damage was observed in the wild type during culture. Scanning and transmission electron microscopy analyses revealed deficiencies of the cell envelope and perforation of the cell wall. Leakage of UV-absorbing substances from the mutants was also observed. Thus, the partial viability of the mutants and their independence of d-alanine for growth indicated that inactivation of alr-2 does not impose an auxotrophic requirement for d-alanine. © FEMS 2015. All rights reserved. For permissions, please e-mail: journals.permissions@oup.com.

A dual-color luciferase assay system reveals circadian resetting of cultured fibroblasts by co-cultured adrenal glands.

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Takako Noguchi

Full Text Available In mammals, circadian rhythms of various organs and tissues are synchronized by pacemaker neurons in the suprachiasmatic nucleus (SCN of the hypothalamus. Glucocorticoids released from the adrenal glands can synchronize circadian rhythms in other tissues. Many hormones show circadian rhythms in their plasma concentrations; however, whether organs outside the SCN can serve as master synchronizers to entrain circadian rhythms in target tissues is not well understood. To further delineate the function of the adrenal glands and the interactions of circadian rhythms in putative master synchronizing organs and their target tissues, here we report a simple co-culture system using a dual-color luciferase assay to monitor circadian rhythms separately in various explanted tissues and fibroblasts. In this system, circadian rhythms of organs and target cells were simultaneously tracked by the green-emitting beetle luciferase from Pyrearinus termitilluminans (ELuc and the red-emitting beetle luciferase from Phrixothrix hirtus (SLR, respectively. We obtained tissues from the adrenal glands, thyroid glands, and lungs of transgenic mice that expressed ELuc under control of the promoter from a canonical clock gene, mBmal1. The tissues were co-cultured with Rat-1 fibroblasts as representative target cells expressing SLR under control of the mBmal1 promoter. Amplitudes of the circadian rhythms of Rat-1 fibroblasts were potentiated when the fibroblasts were co-cultured with adrenal gland tissue, but not when co-cultured with thyroid gland or lung tissue. The phases of Rat-1 fibroblasts were reset by application of adrenal gland tissue, whereas the phases of adrenal gland tissue were not influenced by Rat-1 fibroblasts. Furthermore, the effect of the adrenal gland tissue on the fibroblasts was blocked by application of a glucocorticoid receptor (GR antagonist. These results demonstrate that glucocorticoids are strong circadian synchronizers for fibroblasts and that

Breeding of guinea-pig (Cavia porcellus, L. 1758) for meat production in Butembo, Democratic Republic of Congo : Literature review and breeding scheme

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Sikiminywa, K.-L.; Godeau, Jean-Marie; Nyongombe, U.-F.; Hornick, Jean-Luc

2013-01-01

Providing animal proteins to humans requires choosing the animal species to be raised when access to land is limited. Butembo, is a refuge city of conflicts area in the east of Democratic Republic of Congo, where guinea-pig (Cavia porcellus) is considered as food. Therefore the breeding of this animal is an option to make more meat available. Genetic selection of guinea pigs and popularization of improved farming techniques can contribute to solve problems of food security in the region. The ...

Novel co-culture plate enables growth dynamic-based assessment of contact-independent microbial interactions.

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Thomas J Moutinho

Full Text Available Interactions between microbes are central to the dynamics of microbial communities. Understanding these interactions is essential for the characterization of communities, yet challenging to accomplish in practice. There are limited available tools for characterizing diffusion-mediated, contact-independent microbial interactions. A practical and widely implemented technique in such characterization involves the simultaneous co-culture of distinct bacterial species and subsequent analysis of relative abundance in the total population. However, distinguishing between species can be logistically challenging. In this paper, we present a low-cost, vertical membrane, co-culture plate to quantify contact-independent interactions between distinct bacterial populations in co-culture via real-time optical density measurements. These measurements can be used to facilitate the analysis of the interaction between microbes that are physically separated by a semipermeable membrane yet able to exchange diffusible molecules. We show that diffusion across the membrane occurs at a sufficient rate to enable effective interaction between physically separate cultures. Two bacterial species commonly found in the cystic fibrotic lung, Pseudomonas aeruginosa and Burkholderia cenocepacia, were co-cultured to demonstrate how this plate may be implemented to study microbial interactions. We have demonstrated that this novel co-culture device is able to reliably generate real-time measurements of optical density data that can be used to characterize interactions between microbial species.

Thin-film fixed-bed reactor (TFFBR for solar photocatalytic inactivation of aquaculture pathogen Aeromonas hydrophila

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Khan Sadia J

2012-01-01

Full Text Available Abstract Background Outbreaks of infectious diseases by microbial pathogens can cause substantial losses of stock in aquaculture systems. There are several ways to eliminate these pathogens including the use of antibiotics, biocides and conventional disinfectants, but these leave undesirable chemical residues. Conversely, using sunlight for disinfection has the advantage of leaving no chemical residue and is particularly suited to countries with sunny climates. Titanium dioxide (TiO2 is a photocatalyst that increases the effectiveness of solar disinfection. In recent years, several different types of solar photocatalytic reactors coated with TiO2 have been developed for waste water and drinking water treatment. In this study a thin-film fixed-bed reactor (TFFBR, designed as a sloping flat plate reactor coated with P25 DEGUSSA TiO2, was used. Results The level of inactivation of the aquaculture pathogen Aeromonas hydrophila ATCC 35654 was determined after travelling across the TFFBR under various natural sunlight conditions (300-1200 W m-2, at 3 different flow rates (4.8, 8.4 and 16.8 L h-1. Bacterial numbers were determined by conventional plate counting using selective agar media, cultured (i under conventional aerobic conditions to detect healthy cells and (ii under conditions designed to neutralise reactive oxygen species (agar medium supplemented with the peroxide scavenger sodium pyruvate at 0.05% w/v, incubated under anaerobic conditions, to detect both healthy and sub-lethally injured (oxygen-sensitive cells. The results clearly demonstrate that high sunlight intensities (≥ 600 W m-2 and low flow rates (4.8 L h-1 provided optimum conditions for inactivation of A. hydrophila ATCC 3564, with greater overall inactivation and fewer sub-lethally injured cells than at low sunlight intensities or high flow rates. Low sunlight intensities resulted in reduced overall inactivation and greater sub-lethal injury at all flow rates. Conclusions This

Microplastics as a vector for the transport of the bacterial fish pathogen species Aeromonas salmonicida.

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Viršek, Manca Kovač; Lovšin, Marija Nika; Koren, Špela; Kržan, Andrej; Peterlin, Monika

2017-12-15

Microplastics is widespread in the marine environment where it can cause numerous negative effects. It can provide space for the growth of organisms and serves as a vector for the long distance transfer of marine microorganisms. In this study, we examined the sea surface concentrations of microplastics in the North Adriatic and characterized bacterial communities living on the microplastics. DNA from microplastics particles was isolated by three different methods, followed by PCR amplification of 16S rDNA, clone libraries preparation and phylogenetic analysis. 28 bacterial species were identified on the microplastics particles including Aeromonas spp. and hydrocarbon-degrading bacterial species. Based on the 16S rDNA sequences the pathogenic fish bacteria Aeromonas salmonicida was identified for the first time on microplastics. Because A. salmonicida is responsible for illnesses in fish, it is crucial to get answers if and how microplastics pollution is responsible for spreading of diseases. Copyright © 2017 Elsevier Ltd. All rights reserved.

Staging Scenes of Co-Cultural Communication: Acting out Aspects of Marginalized and Dominant Identities

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Root, Elizabeth

2018-01-01

Courses: Intercultural Communication, Interracial Communication, or an Interpersonal Communication class that covers co-cultural theory. Objectives: Students will be able to demonstrate a practical application of co-cultural theory by creating scenes that illustrate different communicative approaches and desired outcomes based on communication…

Naringenin-responsive riboswitch-based fluorescent biosensor module for Escherichia coli co-cultures.

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Xiu, Yu; Jang, Sungho; Jones, J Andrew; Zill, Nicholas A; Linhardt, Robert J; Yuan, Qipeng; Jung, Gyoo Yeol; Koffas, Mattheos A G

2017-10-01

The ability to design and construct combinatorial synthetic metabolic pathways has far exceeded our capacity for efficient screening and selection of the resulting microbial strains. The need for high-throughput rapid screening techniques is of upmost importance for the future of synthetic biology and metabolic engineering. Here we describe the development of an RNA riboswitch-based biosensor module with dual fluorescent reporters, and demonstrate a high-throughput flow cytometry-based screening method for identification of naringenin over producing Escherichia coli strains in co-culture. Our efforts helped identify a number of key operating parameters that affect biosensor performance, including the selection of promoter and linker elements within the sensor-actuator domain, and the effect of host strain, fermentation time, and growth medium on sensor dynamic range. The resulting biosensor demonstrates a high correlation between specific fluorescence of the biosensor strain and naringenin titer produced by the second member of the synthetic co-culture system. This technique represents a novel application for synthetic microbial co-cultures and can be expanded from naringenin to any metabolite if a suitable riboswitch is identified. The co-culture technique presented here can be applied to a variety of target metabolites in combination with the SELEX approach for aptamer design. Due to the compartmentalization of the two genetic constructs responsible for production and detection into separate cells and application as independent modules of a synthetic microbial co-culture we have subsequently reduced the need for re-optimization of the producer module when the biosensor is replaced or removed. Biotechnol. Bioeng. 2017;114: 2235-2244. © 2017 Wiley Periodicals, Inc. © 2017 Wiley Periodicals, Inc.

Calcitriol enhances fat synthesis factors and calpain activity in co-cultured cells.

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Choi, Hyuck; Myung, Kyuho

2014-08-01

We have conducted an in vitro experiment to determine whether calcitriol can act as a fat synthesizer and/or meat tenderizer when skeletal muscle cells, adipose tissue, and macrophages are co-cultured. When co-cultured, pro-inflammatory cytokine tumor necrosis factor-α (TNF-α) and interleukin-6 (IL-6) expression increased, whereas decreased anti-inflammatory cytokine (IL-10 and IL-15) expression decreased in both C2C12 and 3T3-L1 cells. Calcitriol increased reactive oxygen species (ROS) production in the media. While adiponectin gene expression decreased, leptin, resistin, CCAAT-enhancer-binding protein-beta (C/EBP-β), and peroxisome proliferator-activated receptor gamma (PPAR-γ) gene expression was significantly (P cultured with two different cell types. Inducible nitric oxide synthase (iNOS) protein levels were also stimulated in the C2C12 and 3T3-L1 cells, but arginase l was attenuated by calcitriol. Cacitriol highly amplified (P = 0.008) µ-calpain gene expression in co-cultured C2C12 cells. The results showed an overall increase in pro-inflammatory cytokines and a decrease in anti-inflammatory cytokines of C2C12 and 3T3-L1 cells with calcitriol in co-culture systems. µ-Calpain protein was also augmented in differentiated C2C12 cells with calcitriol. These findings suggest that calcitriol can be used as not only fat synthesizer, but meat tenderizer, in meat-producing animals. © 2014 International Federation for Cell Biology.

Identification of chitinolytic bacteria isolated from shrimp pond sediment and characterization of their chitinase encoding gene

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Triwijayani, A. U.; Puspita, I. D.; Murwantoko; Ustadi

2018-03-01

Chitinolytic bacteria are a group of bacteria owning enzymes that able to hydrolyze chitin. Previously, we isolated chitinolytic bacteria from shrimp pond sediment in Bantul, Yogyakarta, and obtained five isolates showing high chitinolytic index named as isolate PT1, PT2, PT5, PT6 and PB2. The aims of this study were to identify chitinolytic bacteria isolated from shrimp pond sediment and to characterize the chitinase encoding gene from each isolate. The molecular technique was performed by amplification of 16S rDNA, amplification of chitinase encoding gene and sequence analysis. Two chitinolytic bacteria of PT1 and PT2 were similar to Aeromonas bivalvium strain D15, PT5 to Pseudomonas stutzeri strain BD-2.2.1, PT6 to Serratia marcescens strain FZSF02 and PB2 to Streptomyces misionensis strain OsiRt-1. The comparison of chitinase encoding gene between three isolates with those in Gen Bank shows that PT1 had similar sequences with the chi1 gene in Aeromonas sp. 17m, PT2 with chi1 gene in A. caviae (CB101) and PT6 with chiB gene in S. Marcescens (BJL200).

Lactobacillus caviae sp. nov., an obligately heterofermentative bacterium isolated from the oral cavity of a guinea pig (Cavia aperea f. porcellus).

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Killer, Jiri; Pechar, Radko; Švec, Pavel; Salmonová, Hana; Švejstil, Roman; Geigerová, Martina; Rada, Vojtěch; Vlková, Eva; Mekadim, Chahrazed

2017-08-01

A Gram-stain-positive, facultatively anaerobic, and catalase- and oxidase-negative bacterial strain designated MOZM2T, having 98.4 % 16S rRNA gene sequence identity with Lactobacillus reuteri DSM 20016T, was isolated from a swab of the oral cavity of a home-bred guinea pig. Comparative analyses based on the hsp60, pheS and tuf genes confirmed L. reuteri as its closest relative species, with calculated sequence similarities of 92.8, 88.8 and 96.9 %, respectively. DNA-DNA hybridisation revealed a 42 % degree of genetic similarity between the novel strain and L. reuteri DSM 20016T. Strain MOZM2T degrades carbohydrates via the 6-phosphogluconate/phosphoketolase pathway, evidenced by its production of gas from glucose and the end products of hexose catabolism. Comparative analysis of the cellular fatty acid profiles determined significant differences between MOZM2T and L. reuteri DSM 20016T in their proportions of C8 : 0, C14 : 1, C17 : 0, C18 : 2ω6t and C20 : 0 fatty acids. Results of genotypic analyses also demonstrated differences between these two strains. They also differed in DNA G+C content, and some biochemical and physiological characteristics. We therefore believe that the examined bacterial isolate should be considered as a new taxon within the group of obligately heterofermentative lactobacilli. The species name Lactobacillus caviae sp. nov. is proposed, of which the type strain is MOZM2T (=CCM 8609T=DSM 100239T=LMG 28780T).

Boosting dark fermentation with co-cultures of extreme thermophiles for biohythane production from garden waste.

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Abreu, Angela A; Tavares, Fábio; Alves, Maria Madalena; Pereira, Maria Alcina

2016-11-01

Proof of principle of biohythane and potential energy production from garden waste (GW) is demonstrated in this study in a two-step process coupling dark fermentation and anaerobic digestion. The synergistic effect of using co-cultures of extreme thermophiles to intensify biohydrogen dark fermentation is demonstrated using xylose, cellobiose and GW. Co-culture of Caldicellulosiruptor saccharolyticus and Thermotoga maritima showed higher hydrogen production yields from xylose (2.7±0.1molmol(-1) total sugar) and cellobiose (4.8±0.3molmol(-1) total sugar) compared to individual cultures. Co-culture of extreme thermophiles C. saccharolyticus and Caldicellulosiruptor bescii increased synergistically the hydrogen production yield from GW (98.3±6.9Lkg(-1) (VS)) compared to individual cultures and co-culture of T. maritima and C. saccharolyticus. The biochemical methane potential of the fermentation end-products was 322±10Lkg(-1) (CODt). Biohythane, a biogas enriched with 15% hydrogen could be obtained from GW, yielding a potential energy generation of 22.2MJkg(-1) (VS). Copyright © 2016 Elsevier Ltd. All rights reserved.

The biofilteration ability of oysters (Crassostrea gigas) to reduce Aeromonas salmonicida in salmon culture.

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Ma, Xiaona; Li, Xian; Sun, Guoxiang; Sharawy, Zaki Zaki; Qiu, Tianlong; Du, Yishuai; Liu, Ying

2017-07-01

Pathogen contamination in the environment is inevitable with the rapid development of intensive aquaculture. Therefore, alternative ecofriendly biological strategies to control pathogenic bacteria are required. However, our aim was to investigate the ability of oysters (Crassostrea gigas) to filter the important opportunistic pathogen, Aeromonas salmonicida (strain C4), using a green fluorescent protein tag (GFP) in the Atlantic salmon (Salmo salar) farming wastewater. Hence, A. salmonicida removal efficiency and ingestion rate were detected in two different oyster stages (larvae and adults). To evaluate the practical performance of oysters as A. salmonicida biofilter, adult oysters were applied to an integrated constructed wetlands system (ICWS) and their long-term C4-GFP removal efficiency was recorded for 60 days. Overall, our results clearly indicated that oysters had substantial A. salmonicida removal ability via their ingestion process when observed under a fluorescent microscope. Approximately 88-95% of C4-GFP was removed by oyster larvae at an ingestion rate of 6.4 × 10 3 -6.2 × 10 5  CFU/h·ind, while 79-92% of C4-GFP was removed by adult oysters at an ingestion rate of 2.1 × 10 4 -3.1 × 10 6  CFU/h·ind. Furthermore, 57.9 ± 17.2% of C4-GFP removal efficiency was achieved when oysters were applied to ICWS. We, therefore, concluded that using oysters as a biofilter represents an effective alternative for removing A. salmonicida from aquaculture wastewater. However, the fate of oysters after ingesting the pathogenic bacteria, acting as a potential reservoir or vector for pathogens, is still debatable. This research provides the basis for the application of oysters as a biofilter to remove pathogens from aquaculture wastewater in industrialized production.

Modelling the effects of PSII inhibitor pulse exposure on two algae in co-culture.

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Copin, Pierre-Jean; Chèvre, Nathalie

2018-03-01

A weakness of standard testing procedures is that they do not consider interactions between organisms, and they focus only on single species. Furthermore, these procedures do not take into account pulse exposure. However, pulse exposure is of particular importance because in streams, after crop application and during and after precipitation, herbicide concentrations fluctuate widely and can exceed the Annual Average Environmental Quality Standards (AA-EQS), which aim to protect the aquatic environment. The sensitivity of the algae Scenedesmus vacuolatus and Pseudokirchneriella subcapitata in a co-culture exposed to pulses is thus analysed in this study. As a first step, the growths of the algae in co-culture are investigated. For initial cell densities fixed, respectively, to 100,000 and 50,000 cells/mL, the growth of each alga is exponential over at least 48 h. S. vacuolatus seems to influence the growth of P. subcapitata negatively. Allelopathy is a possible explanation for this growth inhibition. The toxicity of the herbicide isoproturon is later tested on the algae S. vacuolatus and P. subcapitata cultured alone and in the co-culture. Despite the supplementary stress on the algae in the co-culture competing for nutrients, the toxicity of the herbicide is lower for the two algae when they are in the co-culture than when they are in separated culture. A model is adapted and used to predict the cell-density inhibition on the alga S. vacuolatus in the co-culture with the alga P. subcapitata exposed to a pulse concentration of isoproturon. Four laboratory experiments are performed to validate the model. The comparison between the laboratory and the modelled effects shows good agreement. The differences can be considered minor most of time. For future studies, it is important to ensure that the cell count is precise, as it is used to determine the parameters of the model. The differences can be also induced by the fact that the cell number of the alga P

A porcine astrocyte/endothelial cell co-culture model of the blood-brain barrier.

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Jeliazkova-Mecheva, Valentina V; Bobilya, Dennis J

2003-10-01

A method for the isolation of porcine atrocytes as a simple extension of a previously described procedure for isolation of brain capillary endothelial cells from adolescent pigs [Methods Cell Sci. 17 (1995) 2] is described. The obtained astroglial culture purified through two passages and by the method of the selective detachment was validated by a phase contrast microscopy and through an immunofluorescent assay for the glial fibrillary acidic protein (GFAP). Porcine astrocytes were co-cultivated with porcine brain capillary endothelial cells (PBCEC) for the development of an in vitro blood-brain barrier (BBB) model. The model was visualized by an electron microscopy and showed elevated transendothellial electrical resistance and reduced inulin permeability. To our knowledge, this is the first report for the establishment of a porcine astrocyte/endothelial cell co-culture BBB model, which avoids interspecies and age differences between the two cell types, usually encountered in the other reported co-culture BBB models. Considering the availability of the porcine brain tissue and the close physiological and anatomical relation between the human and pig brain, the porcine astrocyte/endothelial cell co-culture system can serve as a reliable and easily reproducible model for different in vitro BBB studies.

Semipermeable Capsules Wrapping a Multifunctional and Self-regulated Co-culture Microenvironment for Osteogenic Differentiation

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Correia, Clara R.; Pirraco, Rogério P.; Cerqueira, Mariana T.; Marques, Alexandra P.; Reis, Rui L.; Mano, João F.

2016-02-01

A new concept of semipermeable reservoirs containing co-cultures of cells and supporting microparticles is presented, inspired by the multi-phenotypic cellular environment of bone. Based on the deconstruction of the “stem cell niche”, the developed capsules are designed to drive a self-regulated osteogenesis. PLLA microparticles functionalized with collagen I, and a co-culture of adipose stem (ASCs) and endothelial (ECs) cells are immobilized in spherical liquified capsules. The capsules are coated with multilayers of poly(L-lysine), alginate, and chitosan nano-assembled through layer-by-layer. Capsules encapsulating ASCs alone or in a co-culture with ECs are cultured in endothelial medium with or without osteogenic differentiation factors. Results show that osteogenesis is enhanced by the co-encapsulation, which occurs even in the absence of differentiation factors. These findings are supported by an increased ALP activity and matrix mineralization, osteopontin detection, and the up regulation of BMP-2, RUNX2 and BSP. The liquified co-capsules also act as a VEGF and BMP-2 cytokines release system. The proposed liquified capsules might be a valuable injectable self-regulated system for bone regeneration employing highly translational cell sources.

Cultural impact on co-design teamwork in distributed bi- national teams

NARCIS (Netherlands)

Man, J.; Lu, Y.; Brombacher, A.C.; Ying, F.

2014-01-01

This paper is designed to study the impact of cultural difference on co-design teamwork. The aim of this study is to explore the impact of supporting and hindering cultural factors on codesign teamwork in distributed bi-national teams from Netherland and China. In order to achieve the research

Rabbit uterine epithelial cells: Co-culture with spermatozoa

International Nuclear Information System (INIS)

Boice, M.L.

1988-01-01

A primary culture of rabbit uterine epithelial cells was established and their effects on sperm function were examined in vitro. Epithelial cells were isolated from uteri of estrous rabbits and cultured on floating collagen gels in phenol red-free medium supplemented with 5% fetal bovine serum. Light microscopy and keratin staining showed that the epithelial cell population established in culture had morphological characteristics similar to that seen in the intact endometrium. Cells were cultured with 3 H-leucine and uptake of label by cells and its incorporation into cellular and secretory proteins determined. When compared to cells cultured for 24-48 h, incorporation of label into cellular protein was lower at 72-96 h, but secretion increased. Estradiol 17-β did not affect label uptake or incorporation, but did enhance proliferation of cells as judged by total DNA content of the cell population. Analysis of proteins in media by sodium dodecyl sulfate polyacrylamide gel electrophoresis and fluorography suggested that epithelial and stromal cells synthesis proteins that may be secretory in nature during 72-96 h culture. Twenty-nine to thirty-one h after initiation of epithelial cultures, 1-2 x 10 6 sperm were co-incubated with cells and sperm viability, motility, loss of acrosome and fertilizing ability determined

Treatment of real wastewater using co-culture of immobilized Chlorella vulgaris and suspended activated sludge.

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Mujtaba, Ghulam; Lee, Kisay

2017-09-01

The use of algal-bacterial symbiotic association establishes a sustainable and cost-effective strategy in wastewater treatment. Using municipal wastewater, the removal performances of inorganic nutrients (nitrogen and phosphorus) and organic pollutants were investigated by the co-culture system having different inoculum ratios (R) of suspended activated sludge to alginate-immobilized microalgae Chlorella vulgaris. The co-culture reactors with lower R ratios obtained more removal of nitrogen than in pure culture of C. vulgaris. The reactor with R = 0.5 (sludge/microalgae) showed the highest performance representing 66% removal after 24 h and 95% removal after 84 h. Phosphorus was completely eliminated (100%) in the co-culture system with inoculum ratios of 0.5 and 1.0 after 24 h and in the pure C. vulgaris culture after 36 h. The COD level was greatly reduced in the activated sludge reactor, while, it was increasing in pure C. vulgaris culture after 24 h of incubation. However, COD was almost stabilized after 24 h in the reactors with high R ratios such as 2.0, 5.0, and 10 due to the higher concentration of activated sludge. The growth of C. vulgaris was promoted from 0.03 g/L/d to 0.05 g/L/d in the co-culture of low inoculum ratios such as R = 0.5, implying that there exist an optimum inoculum ratio in the co-culture system in order to achieve efficient removal of nutrients. Copyright © 2017 Elsevier Ltd. All rights reserved.

Immunoglobulin production in human mixed lymphocyte cultures: implications for co-cultures of cells from patients and healthy donors

International Nuclear Information System (INIS)

Ruemke, H.C.; Terpstra, F.G.; Huis, B.; Out, T.A.; Zeijlemaker, W.P.

1982-01-01

When human peripheral blood lymphocytes (PBL) are cultured in the presence of irradiated allogeneic lymphocytes, the resulting mixed lymphocyte reaction (MLR) leads to the secretion into the supernatant of substantial amounts of IgM and IgG, derived from nonirradiated responder B lymphocytes. Our data indicate that stimulation to Ig production by responder B cells may result from different types of of interactions. First, B cells and monocytes among the irradiated stimulator cells activate T responder B cells to produce Ig; second, ''responder'' B cells activate irradiated ''stimulator'' T cells, leading to a ''helper'' signal, back to the responder B cells and leading to Ig production. The latter system is radiosensitive, because allogeneic T cells, irradiated at a dose of 4000 rad or more, failed to induce Ig production by responder B cells. In some combinations of human allogeneic lymphocytes, the co-culture of the cells leads to inhibition of Ig production, both in the presence and in the absence of PWM. Thus, co-culture of allogeneic cells may cause ''positive'' as well as ''negative'' allogeneic effects. The implications of these findings for the interpretation of co-cultures that are aimed at establishing defects in lymphocytes from patients with, for example, immunodeficiencies, who fail to produce Ig in the presence of PWM are discussed

Microbial transformation of tannin-rich substrate to gallic acid through co-culture method.

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Banerjee, Rintu; Mukherjee, Gargi; Patra, Krushna Chandra

2005-05-01

Modified solid-state fermentation (MSSF) of tannin-rich substrate yielding tannase and gallic acid was carried out using a co-culture of the filamentous fungi, Rhizopus oryzae (RO IIT RB-13, NRRL 21498) and Aspergillus foetidus (GMRB013 MTCC 3557). Powdered fruits of Terminalia chebula and powdered pod cover of Caesalpinia digyna was used in the process and the different process parameters for maximum production of tannase and gallic acid by co-culture method were optimized through media engineering. MSSF was carried out at the optimum conditions of 30 degrees C and 80% relative humidity. The optimal pH and incubation period was 5.0 and 48 h respectively. Through the co-culture technique the maximum yield of tannase and gallic acid was found to be 41.3 U/ml and 94.8% respectively.

The effect of combined Moringa oleifera and demineralized freeze-dried bovine bone xenograft on the amount of osteoblast and osteoclast in the healing of tooth extraction socket of Cavia cobaya

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Rostiny Rostiny

2016-12-01

Full Text Available Background: Alveolar bone has an important role in providing support to teeth and dentures. Loss of support caused by alveolar resorption will cause functional and aesthetic problems. Preservation socket using bone graft is one way to maintain the dimensions of the alveolar bone. Moringa oleifera leaf can increase the activity of bone graft in the formation of new bone. Purpose: This study was aimed to evaluate the effect of combined Moringa oleivera leaf extract and demineralized freeze-dried bovine bone xenograft (DFDBBX towards the formation of osteoblasts and osteoclasts in the tooth extraction sockets of cavia cobaya. Method: This study used 28 cavia cobayas divided into four groups. The combination of Moringa oleifera leaf extract and DFDBBX was inducted into the sockets of lower incisor tooth with certain dose in each group, ointment 1 containing PEG (a mixture of PEG 400 and PEG 4000 for control group, ointment 2 containing Moringa oleifera leaf extract and DFDBBX and PEG (at active subtance consentration of 0.5% for group 1, ointment 3 containing Moringa oleifera leaf extract and DFDBBX and PEG (at active substance concentration of 1% for group 2, and Ointment 4 containing Moringa oleifera leaf extract and DFDBBX and PEG (at active substance consentration of 2% for group 3. paraffin block preparations were made for histopathology examination using hematoxylin eosin staining. Result: The results showed that there were significant differences of the number of osteoblasts and osteoclasts in each treatment group (p <0.05. Conclusion: It can be concluded that the combination of Moringa oleifera leaf extract and DFDBBX at 2% cocentration can increase the number of osteoblasts and decrease osteoclasts in the healing of tooth extraction sockets of cavia cobaya.

Necrotizing Fasciitis Caused by Inconspicuous Infection of Aeromonas hydrophila in an Immunocompromised Host

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Kuo-Chun Liao

2010-09-01

Full Text Available We present a case of serious necrotizing fasciitis due to Aeromonas hydrophila without a suggestive history of routes of pathogen invasion in a 60-year-old male. Despite prompt broad-spectrum antibiotic and extensive surgical therapies, the patient died within 72 hours following initial presentation. Our experience suggests clinicians be highly alert to the disease when an immunocompromised patient featured fulminant soft-tissue infection in the endemic area.

Effect of Fibroblast Co-culture on In Vitro Maturation and Fertilization of Mouse Preantral Follicles

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Mahmoud Heidari

2011-01-01

Full Text Available Background: The aim of this study was to evaluate fibroblast co-culture on in vitro maturation andfertilization of prepubertal mouse preantral follicles.Materials and Methods: The ovaries of 12-14 day old mice were dissected and 120-150 μmintact preantral follicles with one or two layers of granulosa cells, and round oocytes were culturedindividually in α-minimal essential medium (α-MEM supplemented with 5% fetal bovine serum(FBS, 100 mIU/ml recombinant follicle stimulating hormone, 1% insulin, transferrin, seleniummix, 100 μg/ml penicillin and 50 μg/ml streptomycin as base medium for 12 days. A total number of226 follicules were cultured under two conditions: i base medium as control group (n=113; ii basemedium co-cultured with mouse embryonic fibroblast (MEF (n=113. Follicular diameters, alone,in addition to other factors were analyzed by student’s t-test and chi-square test, respectively.Results: The co-culture group showed significant differences (p<0.05 in growth rate (days 4, 6 and8 of the culture period and survival rate. However, there was no significant difference in antrumformation, ovulation rate and embryonic development of released oocytes. There were significantdifferences (p<0.05 in the estradiol and progesterone secretion at all days between the co-cultureand control groups.Conclusion: Fibroblast co-culture increased survival rate and steroid production of preantralfollicles by promoting granulosa cell proliferation.

β-Carotene from Yeasts Enhances Laccase Production of Pleurotus eryngii var. ferulae in Co-culture.

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Guo, Chaolin; Zhao, Liting; Wang, Feng; Lu, Jian; Ding, Zhongyang; Shi, Guiyang

2017-01-01

Laccase is widely used in several industrial applications and co-culture is a common method for enhancing laccase production in submerged fermentation. In this study, the co-culture of four yeasts with Pleurotus eryngii var. ferulae was found to enhance laccase production. An analysis of sterilization temperatures and extraction conditions revealed that the stimulatory compound in yeasts was temperature-sensitive, and that it was fat-soluble. An LC-MS analysis revealed that the possible stimulatory compound for laccase production in the four yeast extracts was β-carotene. Moreover, the addition of 4 mg β-carotene to 150 mL of P. eryngii var. ferulae culture broth improved laccase production by 2.2-fold compared with the control (i.e., a monoculture), and was similar to laccase production in co-culture. In addition, the enhanced laccase production was accompanied by an increase of lac gene transcription, which was 6.2-time higher than the control on the fifth day. Therefore, it was concluded that β-carotene from the co-cultured yeasts enhanced laccase production in P. eryngii var. ferulae , and strains that produce β-carotene could be selected to enhance fungal laccase production in a co-culture. Alternatively, β-carotene or crude extracts of β-carotene could be used to induce high laccase production in large scale.

Developing a mesophilic co-culture for direct conversion of cellulose to butanol in consolidated bioprocess.

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Wang, Zhenyu; Cao, Guangli; Zheng, Ju; Fu, Defeng; Song, Jinzhu; Zhang, Junzheng; Zhao, Lei; Yang, Qian

2015-01-01

Consolidated bioprocessing (CBP) of butanol production from cellulosic biomass is a promising strategy for cost saving compared to other processes featuring dedicated cellulase production. CBP requires microbial strains capable of hydrolyzing biomass with enzymes produced on its own with high rate and high conversion and simultaneously produce a desired product at high yield. However, current reported butanol-producing candidates are unable to utilize cellulose as a sole carbon source and energy source. Consequently, developing a co-culture system using different microorganisms by taking advantage of their specific metabolic capacities to produce butanol directly from cellulose in consolidated bioprocess is of great interest. This study was mainly undertaken to find complementary organisms to the butanol producer that allow simultaneous saccharification and fermentation of cellulose to butanol in their co-culture under mesophilic condition. Accordingly, a highly efficient and stable consortium N3 on cellulose degradation was first developed by multiple subcultures. Subsequently, the functional microorganisms with 16S rRNA sequences identical to the denaturing gradient gel electrophoresis (DGGE) profile were isolated from consortium N3. The isolate Clostridium celevecrescens N3-2 exhibited higher cellulose-degrading capability was thus chosen as the partner strain for butanol production with Clostridium acetobutylicum ATCC824. Meanwhile, the established stable consortium N3 was also investigated to produce butanol by co-culturing with C. acetobutylicum ATCC824. Butanol was produced from cellulose when C. acetobutylicum ATCC824 was co-cultured with either consortium N3 or C. celevecrescens N3-2. Co-culturing C. acetobutylicum ATCC824 with the stable consortium N3 resulted in a relatively higher butanol concentration, 3.73 g/L, and higher production yield, 0.145 g/g of glucose equivalent. The newly isolated microbial consortium N3 and strain C. celevecrescens N3

Long-term culture and differentiation of porcine red bone marrow hematopoietic cells co-cultured with immortalized mesenchymal cells.

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Garba, Abubakar; Acar, Delphine D; Roukaerts, Inge D M; Desmarets, Lowiese M B; Devriendt, Bert; Nauwynck, Hans J

2017-09-01

Mesenchymal cells are multipotent stromal cells with self-renewal, differentiation and immunomodulatory capabilities. We aimed to develop a co-culture model for differentiating hematopoietic cells on top of immortalized mesenchymal cells for studying interactions between hematopoietic and mesenchymal cells, useful for adequately exploring the therapeutic potential of mesenchymal cells. In this study, we investigated the survival, proliferation and differentiation of porcine red bone marrow hematopoietic cells co-cultured with immortalized porcine bone marrow mesenchymal cells for a period of five weeks. Directly after collection, primary porcine bone marrow mesenchymal cells adhered firmly to the bottom of the culture plates and showed a fibroblast-like appearance, one week after isolation. Upon immortalization, porcine bone marrow mesenchymal cells were continuously proliferating. They were positive for simian virus 40 (SV40) large T antigen and the mesenchymal cell markers CD44 and CD55. Isolated red bone marrow cells were added to these immortalized mesenchymal cells. Five weeks post-seeding, 92±6% of the red bone marrow hematopoietic cells were still alive and their number increased 3-fold during five weekly subpassages on top of the immortalized mesenchymal cells. The red bone marrow hematopoietic cells were originally small and round; later, the cells increased in size. Some of them became elongated, while others remained round. Tiny dendrites appeared attaching hematopoietic cells to the underlying immortalized mesenchymal cells. Furthermore, weekly differential-quick staining of the cells indicated the presence of monoblasts, monocytes, macrophages and lymphocytes in the co-cultures. At three weeks of co-culture, flow cytometry analysis showed an increased surface expression of CD172a, CD14, CD163, CD169, CD4 and CD8 up to 37±0.8%, 40±8%, 41±4%, 23±3% and 19±5% of the hematopoietic cells, respectively. In conclusion, continuous mesenchymal cell

Bottom-up fabrication of artery-mimicking tubular co-cultures in collagen-based microchannel scaffolds.

Science.gov (United States)

Tan, A; Fujisawa, K; Yukawa, Y; Matsunaga, Y T

2016-10-20

We developed a robust bottom-up approach to construct open-ended, tubular co-culture constructs that simulate the human vascular morphology and microenvironment. By design, these three-dimensional artificial vessels mimic the basic architecture of an artery: a collagen-rich extracellular matrix (as the tunica externa), smooth muscle cells (SMCs) (as the tunica media), and an endothelial cell (EC) lining (as the tunica interna). A versatile needle-based fabrication technique was employed to achieve controllable arterial layouts within a PDMS-hosted collagen microchannel scaffold (330 ± 10 μm in diameter): (direct co-culture) a SMC/EC bilayer to follow the structure of an arteriole-like segment; and (encapsulated co-culture) a lateral SMC multilayer covered by an EC monolayer lining to simulate the architecture of a larger artery. Optical and fluorescence microscopy images clearly evidenced the progressive cell elongation and sprouting behavior of SMCs and ECs along the collagen gel contour and within the gel matrix under static co-culture conditions. The progressive cell growth patterns effectively led to the formation of a tubular co-culture with an internal endothelial lining expressing prominent CD31 (cluster of differentiation 31) intercellular junction markers. During a 4-day static maturation period, the artery constructs showed modest alteration in the luminal diameters (i.e. less than 10% changes from the initial measurements). This argues in favor of stable and predictable arterial architecture achieved via the proposed fabrication protocols. Both co-culture models showed a high glucose metabolic rate during the initial proliferation phase, followed by a temporary quiescent (and thus, mature) stage. These proof-of-concept models with a controllable architecture create an important foundation for advanced vessel manipulations such as the integration of relevant physiological functionality or remodeling into a vascular disease-mimicking tissue.

Co-fermentation of cellobiose and xylose by mixed culture of recombinant Saccharomyces cerevisiae and kinetic modeling.

Science.gov (United States)

Chen, Yingying; Wu, Ying; Zhu, Baotong; Zhang, Guanyu; Wei, Na

2018-01-01

Efficient conversion of cellulosic sugars in cellulosic hydrolysates is important for economically viable production of biofuels from lignocellulosic biomass, but the goal remains a critical challenge. The present study reports a new approach for simultaneous fermentation of cellobiose and xylose by using the co-culture consisting of recombinant Saccharomyces cerevisiae specialist strains. The co-culture system can provide competitive advantage of modularity compared to the single culture system and can be tuned to deal with fluctuations in feedstock composition to achieve robust and cost-effective biofuel production. This study characterized fermentation kinetics of the recombinant cellobiose-consuming S. cerevisiae strain EJ2, xylose-consuming S. cerevisiae strain SR8, and their co-culture. The motivation for kinetic modeling was to provide guidance and prediction of using the co-culture system for simultaneous fermentation of mixed sugars with adjustable biomass of each specialist strain under different substrate concentrations. The kinetic model for the co-culture system was developed based on the pure culture models and incorporated the effects of product inhibition, initial substrate concentration and inoculum size. The model simulations were validated by results from independent fermentation experiments under different substrate conditions, and good agreement was found between model predictions and experimental data from batch fermentation of cellobiose, xylose and their mixtures. Additionally, with the guidance of model prediction, simultaneous co-fermentation of 60 g/L cellobiose and 20 g/L xylose was achieved with the initial cell densities of 0.45 g dry cell weight /L for EJ2 and 0.9 g dry cell weight /L SR8. The results demonstrated that the kinetic modeling could be used to guide the design and optimization of yeast co-culture conditions for achieving simultaneous fermentation of cellobiose and xylose with improved ethanol productivity, which is

The genome of Aeromonas salmonicida subsp. salmonicida A449: insights into the evolution of a fish pathogen

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Murphy Colleen

2008-09-01

Full Text Available Abstract Background Aeromonas salmonicida subsp. salmonicida is a Gram-negative bacterium that is the causative agent of furunculosis, a bacterial septicaemia of salmonid fish. While other species of Aeromonas are opportunistic pathogens or are found in commensal or symbiotic relationships with animal hosts, A. salmonicida subsp. salmonicida causes disease in healthy fish. The genome sequence of A. salmonicida was determined to provide a better understanding of the virulence factors used by this pathogen to infect fish. Results The nucleotide sequences of the A. salmonicida subsp. salmonicida A449 chromosome and two large plasmids are characterized. The chromosome is 4,702,402 bp and encodes 4388 genes, while the two large plasmids are 166,749 and 155,098 bp with 178 and 164 genes, respectively. Notable features are a large inversion in the chromosome and, in one of the large plasmids, the presence of a Tn21 composite transposon containing mercury resistance genes and an In2 integron encoding genes for resistance to streptomycin/spectinomycin, quaternary ammonia compounds, sulphonamides and chloramphenicol. A large number of genes encoding potential virulence factors were identified; however, many appear to be pseudogenes since they contain insertion sequences, frameshifts or in-frame stop codons. A total of 170 pseudogenes and 88 insertion sequences (of ten different types are found in the A. salmonicida genome. Comparison with the A. hydrophila ATCC 7966T genome reveals multiple large inversions in the chromosome as well as an approximately 9% difference in gene content indicating instances of single gene or operon loss or gain. A limited number of the pseudogenes found in A. salmonicida A449 were investigated in other Aeromonas strains and species. While nearly all the pseudogenes tested are present in A. salmonicida subsp. salmonicida strains, only about 25% were found in other A. salmonicida subspecies and none were detected in other

Impact of commercial cigarette smoke condensate on brain tissue co-cultured with astrocytes and blood-brain barrier endothelial cells.

Science.gov (United States)

Lee, Seon-Bong; Kim, Ju-Hyeong; Cho, Myung-Haing; Choe, Eun-Sang; Kim, Kwang-Sik; Shim, Soon-Mi

2017-01-01

The purpose of the current study was to investigate the effect of two commercial cigarette smoke condensates (CCSC) on oxidative stress and cell cytotoxicity in human brain (T98G) or astrocytes (U-373 MG) in the presence of human brain microvascular endothelial cells (HBMEC). Cell viability of mono-culture of T98G or U-373 MG was markedly decreased in a concentration-dependent manner, and T98G was more susceptible than U-373 MG to CCSC exposure. Cytotoxicity was less prominent when T98G was co-cultured with HBMEC than when T98G was co-cultured with U-373 MG. Significant reduction in trans-epithelial electric resistance (TEER), a biomarker of cellular integrity was noted in HBMEC co-cultured with T98G (HBMEC-T98G co-culture) and U-373 MG co-cultured with T98G (U-373 MG-T98G co-culture) after 24 or 48 hr CCSC exposure, respectively. TEER value of U-373 MG co-cultured with T98G (79-84%) was higher than HBMEC co-cultured with T98G (62-63%) within 120-hr incubation with CCSC. Reactive oxygen species (ROS) generated by CCSC in mono-culture of T98G and U-373 MG reached highest levels at 4 and 16 mg/ml, respectively. ROS production by T98G fell when co-cultured with HBMEC or U-373MG. These findings suggest that adverse consequences of CCSC treatment on brain cells may be protected by blood-brain barrier or astrocytes, but with chronic exposure toxicity may be worsened due to destruction of cellular integrity.

Total protein electrophoresis and RAPD fingerprinting analysis for the identification of Aeromonas at the species level Análise eletroforética de proteínas totais e marcadores de RAPD na identificação de Aeromonas ao nível de espécie

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Ana Paula Longaray Delamare

2002-12-01

Full Text Available Fifteen well-defined strains of Aeromonas of thirteen species were analyzed by SDS protein electrophoretic analysis (SDS-PAGE and random amplified polymorphic DNA analysis (RAPD. The comparison between the patterns obtained by both methods allowed differentiating all the strains. Clusters formed by the unweighted pair group method with arithmetic averages applied to protein data correlates with the genetic and biochemical information about the species. The results show that protein fingerprinting has the potential to differentiate Aeromonas species, but the low qualitative variation indicates that this technique is not efficient for the characterization of strains within a species. Conversely, RAPD fingerprinting allows the identification of strains but the high variability limits its potential as an aiding method for species identification.Quinze linhagens de Aeromonas pertencentes a treze espécies foram avaliadas através de eletroforese de proteínas totais (SDS-PAGE e segmentos de DNA amplificados ao acaso (RAPD. A comparação entre os padrões obtidos por ambos métodos permitiu diferenciar todas as linhagens. Agrupamentos formados com base nos dados protéicos mostraram relação com informações bioquímicas e genéticas a respeito das espécies. Os resultados mostraram que análises protéicas têm potencial para diferenciar espécies de Aeromonas, mas a baixa variação qualitativa indica que esta técnica não é eficiente para a caracterização entre linhagens dentro de espécies. Ao contrário, marcadores de RAPD permitem identificar linhagens, mas a alta variabilidade limita seu potencial como método auxiliar na identificação de espécies.

Glia co-culture with neurons in microfluidic platforms promotes the formation and stabilization of synaptic contacts.

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Shi, Mingjian; Majumdar, Devi; Gao, Yandong; Brewer, Bryson M; Goodwin, Cody R; McLean, John A; Li, Deyu; Webb, Donna J

2013-08-07

Two novel microfluidic cell culture schemes, a vertically-layered set-up and a four chamber set-up, were developed for co-culturing central nervous system (CNS) neurons and glia. The cell chambers in these devices were separated by pressure-enabled valve barriers, which permitted us to control communication between the two cell types. The unique design of these devices facilitated the co-culture of glia with neurons in close proximity (∼50-100 μm), differential transfection of neuronal populations, and dynamic visualization of neuronal interactions, such as the development of synapses. With these co-culture devices, initial synaptic contact between neurons transfected with different fluorescent markers, such as green fluorescent protein (GFP) and mCherry-synaptophysin, was imaged using high-resolution fluorescence microscopy. The presence of glial cells had a profound influence on synapses by increasing the number and stability of synaptic contacts. Interestingly, as determined by liquid chromatography-ion mobility-mass spectrometry, neuron-glia co-cultures produced elevated levels of soluble factors compared to that secreted by individual neuron or glia cultures, suggesting a potential mechanism by which neuron-glia interactions could modulate synaptic function. Collectively, these results show that communication between neurons and glia is critical for the formation and stability of synapses and point to the importance of developing neuron-glia co-culture systems such as the microfluidic platforms described in this study.

Teste intradérmico com proteínas recombinantes de Mycobacterium bovis como antígenos em Cavia porcellus

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Elaine S.P. Melo

2014-10-01

Full Text Available O teste intradérmico para o diagnóstico da tuberculose bovina utiliza derivados proteicos purificados (PPD de Mycobacterium bovis que são capazes de induzir reações de hipersensibilidade em animais infectados. No entanto, apresenta baixa especificidade devido à ocorrência de reações cruzadas com outras micobactérias. Neste sentido, o objetivo desse trabalho foi produzir proteínas recombinantes (ESAT-6, PE13, PE5 e ESX-1 de Mycobacterium bovis e avaliá-las como antígenos em teste intradérmico utilizando Cavia porcellus como modelo, e verificar se as condições empregadas na purificação (nativa ou desnaturante interferem no desempenho antigênico dessas proteínas. As proteínas foram testadas em Cavia porcellus previamente sensibilizados com cepa M. bovis AN5 inativada, individualmente (160 µg ou combinadas na forma de um coquetel (40 µg cada. O coquetel de proteínas induziu reações de hipersensibilidade nos animais sensibilizados significativamente superiores (p=0,002 as observadas nos animais não sensibilizados, possibilitando diferenciação. No entanto, as proteínas isoladamente não foram capazes de promover essa diferenciação. As condições de solubilização e purificação influenciaram o desempenho antigênico da proteína ESAT-6, pois, quando produzida em condição desnaturante desencadeou reações inespecíficas nos animais não sensibilizados, enquanto que aquela produzida em condições nativas e aplicada em concentrações de 6, 12, 24 e 48µg induziu reações significativas apenas nos animais sensibilizados, confirmando o seu potencial como antígeno.

Culturable gut microbiota diversity in zebrafish.

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Cantas, Leon; Sørby, Jan Roger Torp; Aleström, Peter; Sørum, Henning

2012-03-01

The zebrafish (Danio rerio) is an increasingly used laboratory animal model in basic biology and biomedicine, novel drug development, and toxicology. The wide use has increased the demand for optimized husbandry protocols to ensure animal health care and welfare. The knowledge about the correlation between culturable zebrafish intestinal microbiota and health in relation to environmental factors and management procedures is very limited. A semi-quantitative level of growth of individual types of bacteria was determined and associated with sampling points. A total of 72 TAB line zebrafish from four laboratories (Labs A-D) in the Zebrafish Network Norway were used. Diagnostic was based on traditional bacterial culture methods and biochemical characterization using commercial kits, followed by 16S rDNA gene sequencing from pure subcultures. Also selected Gram-negative isolates were analyzed for antibiotic susceptibility to 8 different antibiotics. A total of 13 morphologically different bacterial species were the most prevalent: Aeromonas hydrophila, Aeromonas sobria, Vibrio parahaemolyticus, Photobacterium damselae, Pseudomonas aeruginosa, Pseudomonas fluorescens, Pseudomonas luteola, Comamonas testosteroni, Ochrobactrum anthropi, Staphylococcus cohnii, Staphylococcus epidermidis, Staphylococcus capitis, and Staphylococcus warneri. Only Lab B had significantly higher levels of total bacterial growth (OR=2.03), whereas numbers from Lab C (OR=1.01) and Lab D (OR=1.12) were found to be similar to the baseline Lab A. Sexually immature individuals had a significantly higher level of harvested total bacterial growth than mature fish (OR=0.82), no statistically significant differences were found between male and female fish (OR=1.01), and the posterior intestinal segment demonstrated a higher degree of culturable bacteria than the anterior segment (OR=4.1). Multiple antibiotic (>3) resistance was observed in 17% of the strains. We propose that a rapid conventional

Triple co-culture cell model as an in vitro model for oral particulate vaccine systems

DEFF Research Database (Denmark)

Nielsen, Line Hagner; De Rossi, C.; Lehr, C-M.

; this was not observed with ovalbumin and blank solution. An example of the results is shown in Figure 2 for IL-17A. An established co-culture of Caco-2, THP-1 and MUTZ-3 cells showed promise as an in vitro model for testing of oral vaccine formulations. Mobility of co-culture immune cells as well as cytokine production......A triple co-culture cell model of Caco-2 cells, dendritic cells and macrophages (Figure 1) has previously been developed for studying intestinal permeability in a state of inflammation [1],[2]. The aim of this study was to investigate the applicability of this cell model for testing...... the model antigen ovalbumin was spray dried to obtain a particulate vaccine model system for testing in the cell model. The precursors were shown to form cubosomes when dispersed in aqueous medium, and was therefore used as the vaccine formulation for testing on the co-cultures. After 11 days, the TEER...

Phototrophic hydrogen production from glucose by pure and co-cultures of Clostridium butyricum and Rhodobacter sphaeroides

Energy Technology Data Exchange (ETDEWEB)

Fang, Herbert H.P.; Zhu, Heguang; Zhang, Tong [Centre for Environmental Engineering Research, Department of Civil Engineering, University of Hong Kong, Pokfulam Road, Hong Kong (China)

2006-12-15

Phototrophic hydrogen production from glucose by pure and co-cultures of Clostridium butyricum and Rhodobacter sphaeroides was studied in batch experiments. Results showed that in all batches hydrogen was produced after a lag phase of about 10h; pure culture of R. sphaeroides produced hydrogen at rates substantially lower than C. butyricum. In co-culture systems, R. sphaeroides even with cell populations 5.9 times higher still could not compete with C. butyricum for glucose. In co-culture systems, R. sphaeroides syntrophically interacted with C. butyricum, using the acetate and butyrate produced by the latter as substrate for hydrogen production. Hydrogen production was ceased in all batches when the pH was lowered to the level of pH 6.5, resulting from the accumulation of fatty acids. It was also demonstrated in this study that fluorescence in situ hybridization (FISH) was an effective means for the quantification of the relative abundance of individual bacteria in a co-culture system. (author)

Establishment of mouse neuron and microglial cell co-cultured models and its action mechanism.

Science.gov (United States)

Zhang, Bo; Yang, Yunfeng; Tang, Jun; Tao, Yihao; Jiang, Bing; Chen, Zhi; Feng, Hua; Yang, Liming; Zhu, Gang

2017-06-27

The objective of this study is to establish a co-culture model of mouse neurons and microglial cells, and to analyze the mechanism of action of oxygen glucose deprivation (OGD) and transient oxygen glucose deprivation (tOGD) preconditioning cell models. Mouse primary neurons and BV2 microglial cells were successfully cultured, and the OGD and tOGD models were also established. In the co-culture of mouse primary neurons and microglial cells, the cell number of tOGD mouse neurons and microglial cells was larger than the OGD cell number, observed by a microscope. CCK-8 assay result showed that at 1h after treatment, the OD value in the control group is lower compared to all the other three groups (P control group compared to other three groups (P neurons cells were cultured. In the meantime mouse BV2 microglia cells were cultured. Two types of cells were co-cultured, and OGD and tOGD cell models were established. There were four groups in the experiment: control group (OGD), treatment group (tOGD+OGD), placebo group (tOGD+OGD+saline) and minocycline intervention group (tOGD+OGD+minocycline). CCK-8 kit was used to detect cell viability and flow cytometry was used to detect apoptosis. In this study, mouse primary neurons and microglial cells were co-cultured. The OGD and tOGD models were established successfully. tOGD was able to effectively protect neurons and microglial cells from damage, and inhibit the apoptosis caused by oxygen glucose deprivation.

Increased methane production in cyanobacteria and methanogenic microbe co-cultures.

Science.gov (United States)

Yeung, Tracey; Kwan, Matthew; Adler, Lewis; Mills, Toby J; Neilan, Brett A; Conibeer, Gavin; Patterson, Robert

2017-11-01

A novel light-to-bioenergy system produced 3.5 times the baseline methane output using a co-culture of cyanobacteria (Oscillatoria sp.) and a methanogenic microbial community. Analysis of micronutrients in the system during the growth phase indicated that cobalt, iron, nickel and zinc were not appreciably consumed. The stable consumption and return of macronutrients calcium and magnesium were also observed. Essential macronutrients nitrogen, in the form of nitrate, and phosphorus showed no cycling during the growth phase and were depleted at rates of 0.35mg/L/day and 0.40µg/L/day, respectively. Biofilm formation increased the resilience of biomass to bacterial degradation in an anaerobic digester, as shown by viability assays of cyanobacterial biofilms in the co-culture. Copyright © 2017 Elsevier Ltd. All rights reserved.

Aerobic bacteria cultured from the mouth of the American opossum (Didelphis virginiana) with reference to bacteria associated with bite infections.

OpenAIRE

Howell, J M; Dalsey, W C

1990-01-01

The American opossum inflicts bite injuries both when hunted for food and when accidentally provoked when handled in captivity. This study involved aerobically culturing organisms from the mouths of seven wild opossums (Didelphis virginiana). Isolates included streptococci, coagulase-positive and -negative staphylococci, Aeromonas spp., Citrobacter freundii, Eikenella corrodens, and Escherichia coli.

Comparative evaluation of infection methods and environmental factors on challenge success: Aeromonas salmonicida infection in vaccinated rainbow trout

DEFF Research Database (Denmark)

Chettri, Jiwan Kumar; Skov, Jakob; Jaafar, Rzgar M.

2015-01-01

When testing vaccine-induced protection an effective and reliable challenge method is a basic requirement and we here present a comparative study on different challenge methods used for infection of rainbow trout Oncorhynchus mykiss with Aeromonas salmonicida, a bacterial pathogen eliciting...

Ractopamina y nivel de proteína de la dieta, respuesta productiva y características de la carcasa de cuyes (Cavia porcellus)

OpenAIRE

García Leandro, Madeline Victoria

2017-01-01

Universidad Nacional Agraria La Molina. Escuela de Posgrado. Maestría en Nutrición El objetivo del presente trabajo fue determinar el efecto del Clorhidrato de Ractopamina (RAC) y el nivel de proteína de la dieta sobre la respuesta productiva y características de la carcasa de cuyes (Cavia porcellus). Se emplearon 96 cuyes machos de 49 días de edad que se distribuyeron en 24 pozas. Cada poza fue una unidad experimental con cuatro cuyes. Se les administró seis dietas con dos niveles de RAC ...

Engineering systems for the generation of patterned co-cultures for controlling cell-cell interactions.

Science.gov (United States)

Kaji, Hirokazu; Camci-Unal, Gulden; Langer, Robert; Khademhosseini, Ali

2011-03-01

Inside the body, cells lie in direct contact or in close proximity to other cell types in a tightly controlled architecture that often regulates the resulting tissue function. Therefore, tissue engineering constructs that aim to reproduce the architecture and the geometry of tissues will benefit from methods of controlling cell-cell interactions with microscale resolution. We discuss the use of microfabrication technologies for generating patterned co-cultures. In addition, we categorize patterned co-culture systems by cell type and discuss the implications of regulating cell-cell interactions in the resulting biological function of the tissues. Patterned co-cultures are a useful tool for fabricating tissue engineered constructs and for studying cell-cell interactions in vitro, because they can be used to control the degree of homotypic and heterotypic cell-cell contact. In addition, this approach can be manipulated to elucidate important factors involved in cell-matrix interactions. Patterned co-culture strategies hold significant potential to develop biomimetic structures for tissue engineering. It is expected that they would create opportunities to develop artificial tissues in the future. This article is part of a Special Issue entitled Nanotechnologies - Emerging Applications in Biomedicine. 2010 Elsevier B.V. All rights reserved.

Draft genome sequence of Pseudomonas mosselii Gil3, isolated from catfish and antagonistic against hypervirulent Aeromonas hydrophila

Science.gov (United States)

Pseudomonas mosselii Gil3 was isolated from a catfish that survived from lethal challenge with hypervirulent Aeromonas hydrophila (vAh). When assayed in vitro, the bacterium showed antagonism against vAh. Sequence analysis revealed that the genome of P. mosselii Gil3 encodes numerous aromatic metabo...

Molecular Phylogenetics and Temporal Diversification in the Genus Aeromonas Based on the Sequences of Five Housekeeping Genes

Science.gov (United States)

Lorén, J. Gaspar; Farfán, Maribel; Fusté, M. Carmen

2014-01-01

Several approaches have been developed to estimate both the relative and absolute rates of speciation and extinction within clades based on molecular phylogenetic reconstructions of evolutionary relationships, according to an underlying model of diversification. However, the macroevolutionary models established for eukaryotes have scarcely been used with prokaryotes. We have investigated the rate and pattern of cladogenesis in the genus Aeromonas (γ-Proteobacteria, Proteobacteria, Bacteria) using the sequences of five housekeeping genes and an uncorrelated relaxed-clock approach. To our knowledge, until now this analysis has never been applied to all the species described in a bacterial genus and thus opens up the possibility of establishing models of speciation from sequence data commonly used in phylogenetic studies of prokaryotes. Our results suggest that the genus Aeromonas began to diverge between 248 and 266 million years ago, exhibiting a constant divergence rate through the Phanerozoic, which could be described as a pure birth process. PMID:24586399

Effet de Panicum maximum sur la productivité des femelles primipares durant le cycle de reproduction chez le cobaye (Cavia porcellus L.

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Danho, M.

2012-01-01

Full Text Available Effect of Panicum maximum on Productivity of Primiparous Females during Reproduction Cycle in Guinea Pigs (Cavia porcellus L.. In Ivory Coast, Guinea pigs reared for meat (Cavia porcellus L. are mainly fed with Panicum maximum. To assess the effect of the latter during pregnancy and lactation (RC of these animals, primiparous dams were fed ad libitum, Panicum maximum alone during the RC (MOD1 or associated with pellets for rabbits during lactation (MOD2, or associated with pellets for rabbits during the last part of the pregnancy period and the lactation (MOD3, or associated with pellets for rabbits during the entire RC (MOD4. The number of corpora lutea per female was 1.3 ± 0.5 and 2.0 ± 0.0 respectively for MOD1 and MOD4. No pre-embryonic mortality was recorded. The mean weight of the young guinea pigs of MOD1 (54.7 ± 10. g was only 55% of that of MOD4 (98.6 ± 13.6 g. At weaning, the average weight gain of young guinea pigs of MOD1 (40.5 ± 22.2 g represented a third of those obtained with other diets that did not significantly differ. At the end of RC, the weight gain of dams was 17 ± 13.3% for MOD1 compared to 50% for MOD2, MOD3 and MOD4. Feeding Panicum maximum alone induces chronic malnutrition which in turn is responsible of the low ovulation rate and reduced growth in guinea pig breeding.

Transitions from mono- to co- to tri-culture uniquely affect gene expression in breast cancer, stromal, and immune compartments.

Science.gov (United States)

Regier, Mary C; Maccoux, Lindsey J; Weinberger, Emma M; Regehr, Keil J; Berry, Scott M; Beebe, David J; Alarid, Elaine T

2016-08-01

Heterotypic interactions in cancer microenvironments play important roles in disease initiation, progression, and spread. Co-culture is the predominant approach used in dissecting paracrine interactions between tumor and stromal cells, but functional results from simple co-cultures frequently fail to correlate to in vivo conditions. Though complex heterotypic in vitro models have improved functional relevance, there is little systematic knowledge of how multi-culture parameters influence this recapitulation. We therefore have employed a more iterative approach to investigate the influence of increasing model complexity; increased heterotypic complexity specifically. Here we describe how the compartmentalized and microscale elements of our multi-culture device allowed us to obtain gene expression data from one cell type at a time in a heterotypic culture where cells communicated through paracrine interactions. With our device we generated a large dataset comprised of cell type specific gene-expression patterns for cultures of increasing complexity (three cell types in mono-, co-, or tri-culture) not readily accessible in other systems. Principal component analysis indicated that gene expression was changed in co-culture but was often more strongly altered in tri-culture as compared to mono-culture. Our analysis revealed that cell type identity and the complexity around it (mono-, co-, or tri-culture) influence gene regulation. We also observed evidence of complementary regulation between cell types in the same heterotypic culture. Here we demonstrate the utility of our platform in providing insight into how tumor and stromal cells respond to microenvironments of varying complexities highlighting the expanding importance of heterotypic cultures that go beyond conventional co-culture.

Boron nitride nanotube-mediated stimulation of cell co-culture on micro-engineered hydrogels.

Directory of Open Access Journals (Sweden)

Leonardo Ricotti

Full Text Available In this paper, we describe the effects of the combination of topographical, mechanical, chemical and intracellular electrical stimuli on a co-culture of fibroblasts and skeletal muscle cells. The co-culture was anisotropically grown onto an engineered micro-grooved (10 µm-wide grooves polyacrylamide substrate, showing a precisely tuned Young's modulus (∼ 14 kPa and a small thickness (∼ 12 µm. We enhanced the co-culture properties through intracellular stimulation produced by piezoelectric nanostructures (i.e., boron nitride nanotubes activated by ultrasounds, thus exploiting the ability of boron nitride nanotubes to convert outer mechanical waves (such as ultrasounds in intracellular electrical stimuli, by exploiting the direct piezoelectric effect. We demonstrated that nanotubes were internalized by muscle cells and localized in both early and late endosomes, while they were not internalized by the underneath fibroblast layer. Muscle cell differentiation benefited from the synergic combination of topographical, mechanical, chemical and nanoparticle-based stimuli, showing good myotube development and alignment towards a preferential direction, as well as high expression of genes encoding key proteins for muscle contraction (i.e., actin and myosin. We also clarified the possible role of fibroblasts in this process, highlighting their response to the above mentioned physical stimuli in terms of gene expression and cytokine production. Finally, calcium imaging-based experiments demonstrated a higher functionality of the stimulated co-cultures.

Molecular characterization and susceptibility to antimicrobial drugs of isolated bacterials from shrimps (“Litopenaeus vannamei” Caracterização molecular e susceptibilidade aos antimicrobianos de isolados bacterianos de camarões

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Ricardo Castelo Branco Albinati

2010-06-01

Full Text Available The objective was to isolate bacteria from gut of shrimps from Litopenaeus vannamei, by biochemical characterization and molecular identification, inhibition activity in vitro of Bacillus cereus and sensitivity pattern determination. The bacterial species isolated were: Aeromonas caviae (n = 7, Alcaligenes denitrificans (n = 1, Bacillus cereus (n = 1 and Enterobacter spp. (n = 3. Bacillus cereus isolated in this study did not have inhibitory activity to other shrimps isolated bacteria evaluated. In the susceptibility to antimicrobial drug test, it were observed 68,7% to erythromycin, 50% to tetracycline, 81,2% to trimethoprim:sulfamethoxazole, neomycin and estreptomycin, 12,5% to lincomycin and ampicillin, 87,5% to enrofloxacin and nitrofurantoin, 93,7% to ceftriaxone, 100% to norfloxacin and nalidix acid. The characterization molecular is important on identifying the microrganisms studied. The nalidixic acid and norfloxacin are antimicrobial drugs with high sensitivity for bacteria isolated from shrimps.Objetivou-se isolar bactérias provenientes do trato intestinal de camarões da espécie Litopenaeus vannamei, por meio da caracterização bioquímica e molecular, atividade de inibição in vitro do Bacillus cereus e perfil de sensibilidade aos antimicrobianos. As espécies bacterianas identificadas foram Aeromonas caviae (n = 7, Alcaligenes denitrificans (n = 1, Bacillus cereus (n = 1 e Enterobacter spp. (n = 3. Bacillus cereus obtido neste estudo não apresentou atividade de inibição frente às demais bactérias isoladas de camarões. Quanto ao perfil de sensibilidade aos antimicrobianos, foram observados 68,7% de eritromicina, 50% de tetraciclina, 81,2% de sulfametoxazol/trimetoprina, neomicina e estreptomicina, 12,5% de lincomicina e ampicilina, 87,5% de enrofloxacina e nitrofurantoína, 93,7% de ceftriaxona, 100% de norfloxacina e ácido nalidíxico. A caracterização molecular é útil para identificação dos microrganismos estudados

Characterization of fetal growth by repeated ultrasound measurements in the wild guinea pig (Cavia aperea).

Science.gov (United States)

Schumann, K; Guenther, A; Göritz, F; Jewgenow, K

2014-08-01

Fetal growth during pregnancy has previously been studied in the domesticated guinea pig (Cavia aperea f. porcellus) after dissecting pregnant females, but there are no studies describing the fetal growth in their wild progenitor, the wild guinea pig (C aperea). In this study, 50 pregnancies of wild guinea pig sows were investigated using modern ultrasound technique. The two most common fetal growth parameters (biparietal diameter [BPD] and crown-rump-length [CRL]) and uterine position were measured. Data revealed similar fetal growth patterns in the wild guinea pig and domesticated guinea pig in the investigated gestation period, although they differ in reproductive milestones such as gestation length (average duration of pregnancy 68 days), average birth weight, and litter mass. In this study, pregnancy lasted on average 60.2 days with a variance of less than a day (0.96 days). The measured fetal growth parameters are strongly correlated with each (R = 0.91; P guinea pig. Copyright © 2014 Elsevier Inc. All rights reserved.

Decreased hypertrophic differentiation accompanies enhanced matrix formation in co-cultures of outer meniscus cells with bone marrow mesenchymal stromal cells

Science.gov (United States)

2012-01-01

Introduction The main objective of this study was to determine whether meniscus cells from the outer (MCO) and inner (MCI) regions of the meniscus interact similarly to or differently with mesenchymal stromal stem cells (MSCs). Previous study had shown that co-culture of meniscus cells with bone marrow-derived MSCs result in enhanced matrix formation relative to mono-cultures of meniscus cells and MSCs. However, the study did not examine if cells from the different regions of the meniscus interacted similarly to or differently with MSCs. Methods Human menisci were harvested from four patients undergoing total knee replacements. Tissue from the outer and inner regions represented pieces taken from one third and two thirds of the radial distance of the meniscus, respectively. Meniscus cells were released from the menisci after collagenase treatment. Bone marrow MSCs were obtained from the iliac crest of two patients after plastic adherence and in vitro culture until passage 2. Primary meniscus cells from the outer (MCO) or inner (MCI) regions of the meniscus were co-cultured with MSCs in three-dimensional (3D) pellet cultures at 1:3 ratio, respectively, for 3 weeks in the presence of serum-free chondrogenic medium containing TGF-β1. Mono-cultures of MCO, MCI and MSCs served as experimental control groups. The tissue formed after 3 weeks was assessed biochemically, histochemically and by quantitative RT-PCR. Results Co-culture of inner (MCI) or outer (MCO) meniscus cells with MSCs resulted in neo-tissue with increased (up to 2.2-fold) proteoglycan (GAG) matrix content relative to tissues formed from mono-cultures of MSCs, MCI and MCO. Co-cultures of MCI or MCO with MSCs produced the same amount of matrix in the tissue formed. However, the expression level of aggrecan was highest in mono-cultures of MSCs but similar in the other four groups. The DNA content of the tissues from co-cultured cells was not statistically different from tissues formed from mono-cultures of

Study on O2 generation and CO2 absorption capability of four co-cultured salad plants in an enclosed system

Science.gov (United States)

Guo, Shuangsheng; Ai, Weidang; Tang, Yongkang; Cheng, Quanyong; Shen, Yunze; Qin, Lifeng; Ma, Jialu; Zhu, Jingtao; Ren, Jin

2014-06-01

The ability to generate O2 and absorb CO2 of several co-cultured vegetable plants in an enclosed system was studied to provide theoretical reference for the future man-plant integrated tests. Four kinds of salad plants (Lactuca sativa L. var. Dasusheng, Lactuca sativa L. var. Youmaicai, Gynura bicolor and Cichorium endivia L.) were grown in the CELSS Integration Test Platform (CITP). The environmental factors including O2 and CO2 concentration were continuously monitored on-line and the plant biomass was measured at the end of the test. The changing rules of O2 and CO2 concentration in the system were basically understood and it was found that the O2 generated by the plants could satisfy the respiratory needs of 1.75 persons by calculation. It was also found that the plants could absorb the CO2 breathed out by 2 persons when the light intensity was raised to 550 mmol m-2 s-1 PPF. The results showed that the co-cultured plants hold good compatibility and excellent O2-generating and CO2-absorbing capability. They could also supply some fresh edible vegetable for a 2-person crew.

Huperzine A protects neural stem cells against Aβ-induced apoptosis in a neural stem cells and microglia co-culture system

Science.gov (United States)

Zhu, Ning; Lin, Jizong; Wang, Kewan; Wei, Meidan; Chen, Qingzhuang; Wang, Yong

2015-01-01

Objectives: This study aims to explore whether Huperzine A (HupA) could protect neural stem cells against amyloid beta-peptide Aβ induced apoptosis in a neural stem cells (NSCs) and microglia co-culture system. Methods: Rat NSCs and microglial cells were isolated, cultured and identified with immunofluorescence Assays (IFA). Co-culture systems of NSCs and microglial cells were employed using Transwell Permeable Supports. The effects of Aβ1-42 on NSCs were studied in 4 groups using co-culture systems: NSCs, Aβ+NSCs, co-culture and Aβ+co-culture groups. Bromodeoxyuridine (BrdU) incorporation and flow cytometry were utilized to assess the differences of proliferation, differentiation and apoptosis of NSCs between the groups. LQ test was performed to assess the amounts of IL-6, TNF-α and MIP-α secreted, and flow cytometry and Western blotting were used to assess apoptosis of NSCs and the expressions of Bcl-2 and Bax in each group. Results: IFA results showed that isolated rat NSCs were nestin-positive and microglial cells were CD11b/c-positive. Among all the groups, the Aβ+co-culture group has the lowest BrdU expression level, the lowest MAP2-positive, ChAT-positive cell counts and the highest NSC apoptosis rate. Smaller amounts of IL-6, TNF-α and MIP-α were being secreted by microglial cells in the HupA+Aβ+co-culture group compared with those in the Aβ+ co-culture group. Also the Bcl-2: Bax ratio was much higher in the HupA+Aβ+co-culture group than in the Aβ+co-culture group. Conclusions: HupA inhibits cell apoptosis through restraining microglia’s inflammatory response induced by Aβ1-42. PMID:26261518

Aerobic bacteria cultured from the mouth of the American opossum (Didelphis virginiana) with reference to bacteria associated with bite infections.

Science.gov (United States)

Howell, J M; Dalsey, W C

1990-01-01

The American opossum inflicts bite injuries both when hunted for food and when accidentally provoked when handled in captivity. This study involved aerobically culturing organisms from the mouths of seven wild opossums (Didelphis virginiana). Isolates included streptococci, coagulase-positive and -negative staphylococci, Aeromonas spp., Citrobacter freundii, Eikenella corrodens, and Escherichia coli. PMID:2229365

Monitoring Dynamic Interactions between Breast Cancer Cells and Human Bone Tissue in a Co-Culture Model

Science.gov (United States)

Contag, Christopher H.; Lie, Wen-Rong; Bammer, Marie C.; Hardy, Jonathan W.; Schmidt, Tobi L.; Maloney, William J.; King, Bonnie L.

2015-01-01

Purpose Bone is a preferential site of breast cancer metastasis and models are needed to study this process at the level of the microenvironment. We have used bioluminescence imaging (BLI) and multiplex biomarker immunoassays to monitor dynamic breast cancer cell behaviors in co-culture with human bone tissue. Procedures Femur tissue fragments harvested from hip replacement surgeries were co-cultured with luciferase-positive MDA-MB-231-fLuc cells. BLI was performed to quantify breast cell division and track migration relative to bone tissue. Breast cell colonization of bone tissues was assessed with immunohistochemistry. Biomarkers in co-culture supernatants were profiled with MILLIPLEX® immunoassays. Results BLI demonstrated increased MDA-MB-231-fLuc proliferation (pbones, and revealed breast cell migration toward bone. Immunohistochemistry illustrated MDA-MB-231-fLuc colonization of bone, and MILLIPLEX® profiles of culture supernatants suggested breast/bone crosstalk. Conclusions Breast cell behaviors that facilitate metastasis occur reproducibly in human bone tissue co-cultures and can be monitored and quantified using BLI and multiplex immunoassays. PMID:24008275

Air-dust-borne associations of phototrophic and hydrocarbon-utilizing microorganisms: promising consortia in volatile hydrocarbon bioremediation.

Science.gov (United States)

Al-Bader, Dhia; Eliyas, Mohamed; Rayan, Rihab; Radwan, Samir

2012-11-01

Aquatic and terrestrial associations of phototrophic and heterotrophic microorganisms active in hydrocarbon bioremediation have been described earlier. The question arises: do similar consortia also occur in the atmosphere? Dust samples at the height of 15 m were collected from Kuwait City air, and analyzed microbiologically for phototrophic and heterotrophic hydrocarbon-utilizing microorganisms, which were subsequently characterized according to their 16S rRNA gene sequences. The hydrocarbon utilization potential of the heterotrophs alone, and in association with the phototrophic partners, was measured quantitatively. The chlorophyte Gloeotila sp. and the two cyanobacteria Nostoc commune and Leptolyngbya thermalis were found associated with dust, and (for comparison) the cynobacteria Leptolyngbya sp. and Acaryochloris sp. were isolated from coastal water. All phototrophic cultures harbored oil vapor-utilizing bacteria in the magnitude of 10(5) g(-1). Each phototrophic culture had its unique oil-utilizing bacteria; however, the bacterial composition in Leptolyngbya cultures from air and water was similar. The hydrocarbon-utilizing bacteria were affiliated with Acinetobacter sp., Aeromonas caviae, Alcanivorax jadensis, Bacillus asahii, Bacillus pumilus, Marinobacter aquaeolei, Paenibacillus sp., and Stenotrophomonas maltophilia. The nonaxenic cultures, when used as inocula in batch cultures, attenuated crude oil in light and dark, and in the presence of antibiotics and absence of nitrogenous compounds. Aqueous and diethyl ether extracts from the phototrophic cultures enhanced the growth of the pertinent oil-utilizing bacteria in batch cultures, with oil vapor as a sole carbon source. It was concluded that the airborne microbial associations may be effective in bioremediating atmospheric hydrocarbon pollutants in situ. Like the aquatic and terrestrial habitats, the atmosphere contains dust-borne associations of phototrophic and heterotrophic hydrocarbon

Susceptibilities to carbapenems and presence of cphA gene on food-borne Aeromonas

Directory of Open Access Journals (Sweden)

Bibiana María Martín Talavera

2006-07-01

Full Text Available The purpose of this study was to determine the susceptibilities of food-borne Aeromonas to carbapenems, as well as to investigate the presence of a metallo carbapenemase-encoding gene, named cphA. Minimum Inhibitory Concentration (MIC was determined following NCCLS standards. All the tested microorganisms were susceptible to imipenem, meropenem and biapenem. However, a strong inoculum size effect on carbapenem MICs was observed for most of the strains. Six strains, out of seven, showed the presence of metallo--beta-lactamases but cphA gene was detected in only two strains of A. veronii bv. sobria.O objetivo deste estudo foi determinar a suscetibilidade de aeromonas de origem alimentar a carbapenems bem como investigar a presença de um gene codificante de metalocarbapenemase, denominado "cph A". A suscetibilidade in vitro foi determinada pelo metodo de diluição em agar. Todas as cepas foram suscetíveis a Imipenem, Meropenem e Biapenem. Porém foi observado um forte efeito de tamanho do inóculo sobre as CIM das carbapenems na maioria das cepas. A detecção de metalo-beta-lactamase foi realizada pelo metodo lodometrico. Seis cepas das sete testadas demostraron a presença da enzima. A presença do gene cphA foi determinada por PCR e foi detectada em duas cepas de A veronii bv. sobria.

Metabolic cooperation between co-cultured lung cancer cells and lung fibroblasts.

Science.gov (United States)

Koukourakis, Michael I; Kalamida, Dimitra; Mitrakas, Achilleas G; Liousia, Maria; Pouliliou, Stamatia; Sivridis, Efthimios; Giatromanolaki, Alexandra

2017-11-01

Cooperation of cancer cells with stromal cells, such as cancer-associated fibroblasts (CAFs), has been revealed as a mechanism sustaining cancer cell survival and growth. In the current study, we focus on the metabolic interactions of MRC5 lung fibroblasts with lung cancer cells (A549 and H1299) using co-culture experiments and studying changes of the metabolic protein expression profile and of their growth and migration abilities. Using western blotting, confocal microscopy and RT-PCR, we observed that in co-cultures MRC5 respond by upregulating pyruvate dehydrogenase (PDH) and the monocarboxylate transporter MCT1. In contrast, cancer cells increase the expression of glucose transporters (GLUT1), LDH5, PDH kinase and the levels of phosphorylated/inactivated pPDH. H1299 cells growing in the same culture medium with fibroblasts exhibit a 'metastasis-like' phenomenon by forming nests within the fibroblast area. LDH5 and pPDH were drastically upregulated in these nests. The growth rate of both MRC5 and cancer cells increased in co-cultures. Suppression of LDHA or PDK1 in cancer cells abrogates the stimulatory signal from cancer cells to fibroblasts. Incubation of MRC5 fibroblasts with lactate resulted in an increase of LDHB and of PDH expression. Silencing of PDH gene in fibroblasts, or silencing of PDK1 or LDHA gene in tumor cells, impedes cancer cell's migration ability. Overall, a metabolic cooperation between lung cancer cells and fibroblasts has been confirmed in the context of direct Warburg effect, thus the fibroblasts reinforce aerobic metabolism to support the intensified anaerobic glycolytic pathways exploited by cancer cells.

Antitumor Activity of Rat Mesenchymal Stem Cells during Direct or Indirect Co-Culturing with C6 Glioma Cells.

Science.gov (United States)

Gabashvili, A N; Baklaushev, V P; Grinenko, N F; Mel'nikov, P A; Cherepanov, S A; Levinsky, A B; Chehonin, V P

2016-02-01

The tumor-suppressive effect of rat mesenchymal stem cells against low-differentiated rat C6 glioma cells during their direct and indirect co-culturing and during culturing of C6 glioma cells in the medium conditioned by mesenchymal stem cells was studied in an in vitro experiment. The most pronounced antitumor activity of mesenchymal stem cells was observed during direct co-culturing with C6 glioma cells. The number of live C6 glioma cells during indirect co-culturing and during culturing in conditioned medium was slightly higher than during direct co-culturing, but significantly differed from the control (C6 glioma cells cultured in medium conditioned by C6 glioma cells). The cytotoxic effect of medium conditioned by mesenchymal stem cells was not related to medium depletion by glioma cells during their growth. The medium conditioned by other "non-stem" cells (rat astrocytes and fibroblasts) produced no tumor-suppressive effect. Rat mesenchymal stem cells, similar to rat C6 glioma cells express connexin 43, the main astroglial gap junction protein. During co-culturing, mesenchymal stem cells and glioma C6 cells formed functionally active gap junctions. Gap junction blockade with connexon inhibitor carbenoxolone attenuated the antitumor effect observed during direct co-culturing of C6 glioma cells and mesenchymal stem cells to the level produced by conditioned medium. Cell-cell signaling mediated by gap junctions can be a mechanism of the tumor-suppressive effect of mesenchymal stem cells against C6 glioma cells. This phenomenon can be used for the development of new methods of cell therapy for high-grade malignant gliomas.

Inventarisatie van Aeromonas spp. in het drinkwater van 21 pompstations in de periode september 1985-december 1986 (deel I)

NARCIS (Netherlands)

Versteegh JFM; During M; Havelaar AH

1987-01-01

In de periode september 1985-december 1986 is in samenwerking met een aantal drinkwaterbedrijven een inventariserend onderzoek uitgevoerd naar het voorkomen van Aeromonas-bacterien in verschillende typen drinkwater geproduceerd op 21 drinkwaterpompstations. Nadat in de loop van het onderzoek

Inventarisatie van Aeromonas spp. in het drinkwater van 21 pompstations in de periode september 1985-december 1986 (deel II)

NARCIS (Netherlands)

Versteegh JFM; During M; Havelaar AH

1987-01-01

In de periode september 1985-december 1986 is in samenwerking met een aantal drinkwaterbedrijven een inventariserend onderzoek uitgevoerd naar het voorkomen van Aeromonas-bacterien in verschillende typen drinkwater geproduceerd op 21 drinkwaterpompstations. Nadat in de loop van het onderzoek

Co-culture of chondrons and mesenchymal stromal cells reduces the loss of collagen VI and improves extracellular matrix production.

Science.gov (United States)

Owida, H A; De Las Heras Ruiz, T; Dhillon, A; Yang, Y; Kuiper, N J

2017-12-01

Adult articular chondrocytes are surrounded by a pericellular matrix (PCM) to form a chondron. The PCM is rich in hyaluronan, proteoglycans, and collagen II, and it is the exclusive location of collagen VI in articular cartilage. Collagen VI anchors the chondrocyte to the PCM. It has been suggested that co-culture of chondrons with mesenchymal stromal cells (MSCs) might enhance extracellular matrix (ECM) production. This co-culture study investigates whether MSCs help to preserve the PCM and increase ECM production. Primary bovine chondrons or chondrocytes or rat MSCs were cultured alone to establish a baseline level for ECM production. A xenogeneic co-culture monolayer model using rat MSCs (20, 50, and 80%) was established. PCM maintenance and ECM production were assessed by biochemical assays, immunofluorescence, and histological staining. Co-culture of MSCs with chondrons enhanced ECM matrix production, as compared to chondrocyte or chondron only cultures. The ratio 50:50 co-culture of MSCs and chondrons resulted in the highest increase in GAG production (18.5 ± 0.54 pg/cell at day 1 and 11 ± 0.38 pg/cell at day 7 in 50:50 co-culture versus 16.8 ± 0.61 pg/cell at day 1 and 10 ± 0.45 pg/cell at day 7 in chondron monoculture). The co-culture of MSCs with chondrons appeared to decelerate the loss of the PCM as determined by collagen VI expression, whilst the expression of high-temperature requirement serine protease A1 (HtrA1) demonstrated an inverse relationship to that of the collagen VI. Together, this implies that MSCs directly or indirectly inhibited HtrA1 activity and the co-culture of MSCs with chondrons enhanced ECM synthesis and the preservation of the PCM.

Characterization of A Three-Dimensional Organotypic Co-Culture Skin Model for Epidermal Differentiation of Rat Adipose-Derived Stem Cells.

Science.gov (United States)

Ghanavati, Zeinab; Orazizadeh, Mahmoud; Bayati, Vahid; Abbaspour, Mohammad Reza; Khorsandi, Layasadat; Mansouri, Esrafil; Neisi, Niloofar

2016-01-01

The organotypic co-culture is a well-known technique to examine cellular interactions and their roles in stem cell proliferation and differentiation. This study aims to evaluate the effects of dermal fibroblasts (DFs) on epidermal differentiation of adipose-derived stem cells (ASCs) using a three-dimensional (3D) organotypic co- culture technique. In this experimental research study, rat DFs and ASCs were isolated and cultured separately on electrospun polycaprolactone (PCL) matrices. The PCL matrices seeded by ASCs were superimposed on to the matrices seeded by DFs in order to create a 3D organotypic co-culture. In the control groups, PCL matrices seeded by ASCs were placed on matrices devoid of DFs. After 10 days, we assessed the expressions of keratinocyte-related genes by real-time reverse transcriptase-polymerase chain reaction (RT-PCR) and expression of pan-cytokeratin protein by immunofluorescence in the differentiated keratinocyte-like cells from co- culture and control groups. Keratinocyte-like cell morphologies were also observed by scanning electron microscopy (SEM). The early, intermediate, and terminal differentiation keratinocyte markers-Cytokeratin14, Filaggrin, and Involucrin significantly expressed in the co-culture groups com- pared to the control ones (P<0.05). We observed pan-cytokeratin in keratinocyte-like cells of both groups by immunofluorescence. SEM observation of the co-culture groups showed that the differentiated keratinocyte-like cells developed a polygonal cobblestone shape, considered characteristic of keratinocytes. The 3D organotypic co-culture bilayered construct that consisted of DFs and ASCs was an effective technique for epidermal differentiation of ASCs. This co-culture might be useful for epidermal differentiation of stem cells for future applications in skin regeneration.

Effects of testicular irradiation with 60Co in guinea pigs CAVIA PORCELLUS

International Nuclear Information System (INIS)

Santos, O.R. dos.

1980-01-01

Improvements were made in restaint systems and in the technique of electroejaculation in guinea pigs, emphasizing safety, effectiveness and repetition. Arrangements for simultaneous testicular irradiation with 60 Co in 5 animals and a phantom to test dose levels in the target region and scattering were also made. Effects of single and fractionated exposures on sperm were studied. A collimated point-shaped 60 Co-machine calibrated to 7 R/min was used as radiation source. Groups of five 12 month aged male guinea pigs were exposed to either single testicular irradiation of 50, 100, 200 and 400 R or fractionated weekly testicular irradiation of 2, 5, 10 and 20 R, during 10 weeks. A last group was exposed to 4 fractionated testicular irradiations of 20 R at time intervals of 21 days and a group of 8 animals was used as control. Semen samples evaluations were made during 5 weeks before irradiation and in the subsequent 11 weeks. Abnormalities in sperm morphology were studied by William's method and phase contrast microscopy. Mean percentages of semen normal espermatzoons and spermatic abnormalities are shown in figures and photomicrographs. Marked differences on concentration, sperm motility and sperm abnormalities were found, but not on volume, in favour of 100 and 200 R single doses levels, as compared to fractionated 10 and 20 R. (Author) [pt

Exploration of Yeast and Bacteria Contaminants in Seed Culture and Fermented Wort from Gyo Gon Alcohol Distillery

International Nuclear Information System (INIS)

Ngwe Thein

2005-10-01

Study was made on samples of seed culture and fermented wort from Gyogon alcohol distillery. In all samples bacteria contaminants were observed. Samples were cultured on Sabouraud dextrose agar, Czapek Dox agar, and nutrient agar media and broth. The selected colonies were isolated. Biochemical tests for identification were conducted. The yeast and bacteria contaminants were identified by morphological characteristics and biochemical reactions. The yeast isolated and identified from Gyogon alcohol distillery was Sacchacromyces cerevisiae. The bacteria contaminants isolated and identified were Aeromonas sp. and Pseudomonas sp.

Antibiotic and heavy metal resistance of Aeromonas hydrophila and Edwardsiella tarda isolated from red hybrid tilapia (Oreochromis spp. coinfected with motile aeromonas septicemia and edwardsiellosis

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S. W. Lee

2017-07-01

Full Text Available Aim: The aim of this study is to identify antibiogram and heavy metal resistance pattern of Aeromonas hydrophila and Edwardsiella tarda isolated from red hybrid tilapia (Oreochromis spp. coinfected with motile aeromonas septicemia and edwardsiellosis in four commercial fish farms. Materials and Methods: A. hydrophila and E. tarda were isolated using glutamate starch phenol red and xylose lysine deoxycholate (Merck, Germany as a selective medium, respectively. All the suspected bacterial colonies were identified using conventional biochemical tests and commercial identification kit (BBL Crystal, USA. Susceptibility testing of present bacterial isolates to 16 types of antibiotics (nalidixic acid, oxolinic acid, compound sulfonamides, doxycycline, tetracycline, novobiocin, chloramphenicol, kanamycin, sulfamethoxazole, flumequine, erythromycin, ampicillin, spiramycin, oxytetracycline, amoxicillin, and fosfomycin and four types of heavy metals (mercury, chromium, copper, and zinc were carried out using disk diffusion and two-fold agar dilution method, respectively. Results: Three hundred isolates of A. hydrophila and E. tarda were successfully identified by biochemical tests. Antibiotic susceptibility testing results showed that 42.2% of the bacterial isolates were sensitive to compound sulfonamides, sulfamethoxazole, flumequine, oxytetracycline, doxycycline, and oxolinic acid. On the other hand, 41.6% of these isolates were resistant to novobiocin, ampicillin, spiramycin, and chloramphenicol, which resulted for multiple antibiotic resistance index values 0.416. Among tested heavy metals, bacterial isolates exhibited resistant pattern of Zn2+ > Cr6+ > Cu2+ > Hg2+. Conclusion: Results from this study indicated that A. hydrophila and E. tarda isolated from coinfected farmed red hybrid tilapia were multi-resistant to antibiotics and heavy metals. These resistant profiles could be useful information to fish farmers to avoid unnecessary use of

Congenital malformations caused by Stryphnodendron fissuratum (Leg. Mimosoideae) in guinea pigs (Cavia porcellus).

Science.gov (United States)

Macedo, Josenaldo S; Rocha, Brena P; Colodel, Edson M; Freitas, Sílvio H; Dória, Renata G S; Riet-Correa, Franklin; Evêncio-Neto, Joaquim; Mendonça, Fábio S

2015-11-01

The aim of this study was to evaluate the toxicity of Stryphnodendron fissuratum pods in guinea pigs (Cavia porcellus) and test the hypothesis that this plant has teratogenic effects. Thus, sixteen guinea pigs were randomly divided into four groups of four animals each. Groups 10, 20 and 40 consisted of guinea pigs that received commercial food that contained crushed pods of S. fissuratum at concentrations of 10, 20 and 40 g/kg, respectively, during the period of organogenesis. Control group consisted of guinea pigs under the same management conditions that did not receive crushed pods of S. fissuratum in their food. In all experimental groups, the main clinical signs of poisoning consisted of anorexia, prostration, absence of vocalizations, alopecia, diarrhea, and abortions within the adult guinea pigs. Those that did not abort gave birth to weak, malnourished pups, some of which had fetal malformations. The main teratogenic changes consisted of eventration, arthrogryposis, amelia of the forelimbs, anophthalmia, microphthalmia, anotia and agnathia. The reductions in the number of offspring and the malformations observed in the experimental groups suggest that S. fissuratum affects fetal development and is teratogenic. Copyright © 2015 Elsevier Ltd. All rights reserved.

A Novel Miniature Culture System to Screen CO2-Sequestering Microalgae

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Xiaoling Miao

2012-11-01

Full Text Available In this study, a novel 96-well microplate swivel system (M96SS was built for high-throughput screening of microalgal strains for CO2 fixation. Cell growth under different CO2 supply conditions (0.2, 0.4, 0.8, and 1.2 g L−1 d−1, residual nitrate, and pH value of Chlorella sp. SJTU-3, Chlorella pyrenoidosa SJTU-2, and Scenedesmus obliquus SJTU-3 were examined in the M96SS and traditional flask cultures. The dynamic data showed there was a good agreement between the systems. Two critical problems in miniature culture systems (intra-well mixing and evaporation loss were improved by sealed vertical mixing of the M96SS. A sample screen of six microalgal species (Chlorella sp. SJTU-3, Chlorella pyrenoidosa SJTU-2, Selenastrum capricornutum, Scenedesmus obliquus SJTU-3, Chlamydomonas sajao, Dunaliella primolecta was carried out in flasks and the M96SS. Chlamydomonas sajao appeared to be a robust performer (highest cell density: 1.437 g L−1 in anaerobic pond water with 0.8, and 1.2 g L−1 d−1 CO2. The reliability and efficiency of the M96SS were verified through a comparison of traditional flask culture, M96SS, Lukavský’s system, and a microplate shaker.

Practices of shake-flask culture and advances in monitoring CO2 and O2.

Science.gov (United States)

Takahashi, Masato; Aoyagi, Hideki

2018-05-01

About 85 years have passed since the shaking culture was devised. Since then, various monitoring devices have been developed to measure culture parameters. O 2 consumed and CO 2 produced by the respiration of cells in shaking cultures are of paramount importance due to their presence in both the culture broth and headspace of shake flask. Monitoring in situ conditions during shake-flask culture is useful for analysing the behaviour of O 2 and CO 2 , which interact according to Henry's law, and is more convenient than conventional sampling that requires interruption of shaking. In situ monitoring devices for shake-flask cultures are classified as direct or the recently developed bypass type. It is important to understand the characteristics of each type along with their unintended effect on shake-flask cultures, in order to improve the existing devices and culture conditions. Technical developments in the bypass monitoring devices are strongly desired in the future. It is also necessary to understand the mechanism underlying conventional shake-flask culture. The existing shaking culture methodology can be expanded into next-generation shake-flask cultures constituting a novel culture environment through a judicious selection of monitoring devices depending on the intended purpose of shake-flask culture. Construction and sharing the databases compatible with the various types of the monitoring devices and measurement instruments adapted for shaking culture can provide a valuable resource for broadening the application of cells with shake-flask culture.

Human nasal turbinates as a viable source of respiratory epithelial cells using co-culture system versus dispase-dissociation technique.

Science.gov (United States)

Noruddin, Nur Adelina Ahmad; Saim, Aminuddin B; Chua, Kien Hui; Idrus, Ruszymah

2007-12-01

To compare a co-culture system with a conventional dispase-dissociation method for obtaining functional human respiratory epithelial cells from the nasal turbinates for tissue engineering application. Human respiratory epithelial cells were serially passaged using a co-culture system and a conventional dispase-dissociation technique. The growth kinetics and gene expression levels of the cultured respiratory epithelial cells were compared. Four genes were investigated, namely cytokeratin-18, a marker for ciliated and secretory epithelial cells; cytokeratin-14, a marker for basal epithelial cells; MKI67, a proliferation marker; and MUC5B, a marker for mucin secretion. Immunocytochemical analysis was performed using monoclonal antibodies against the high molecular-weight cytokeratin 34 beta E12, cytokeratin 18, and MUC5A to investigate the protein expression from cultured respiratory epithelial cells. Respiratory epithelial cells cultured using both methods maintained polygonal morphology throughout the passages. At passage 1, co-cultured respiratory epithelial showed a 2.6-times higher growth rate compared to conventional dispase dissociation technique, and 7.8 times higher at passage 2. Better basal gene expression was observed by co-cultured respiratory epithelial cells compared to dispase dissociated cells. Immunocytochemical analyses were positive for the respiratory epithelial cells cultured using both techniques. Co-culture system produced superior quality of cultured human respiratory epithelial cells from the nasal turbinates as compared to dispase dissociation technique.

In vitro co-cultures of Pinus pinaster with Bursaphelenchus xylophilus: a biotechnological approach to study pine wilt disease.

Science.gov (United States)

Faria, Jorge M S; Sena, Inês; Vieira da Silva, Inês; Ribeiro, Bruno; Barbosa, Pedro; Ascensão, Lia; Bennett, Richard N; Mota, Manuel; Figueiredo, A Cristina

2015-06-01

Co-cultures of Pinus pinaster with Bursaphelenchus xylophilus were established as a biotechnological tool to evaluate the effect of nematotoxics addition in a host/parasite culture system. The pinewood nematode (PWN), Bursaphelenchus xylophilus, the causal agent of pine wilt disease (PWD), was detected for the first time in Europe in 1999 spreading throughout the pine forests in Portugal and recently in Spain. Plant in vitro cultures may be a useful experimental system to investigate the plant/nematode relationships in loco, thus avoiding the difficulties of field assays. In this study, Pinus pinaster in vitro cultures were established and compared to in vivo 1 year-old plantlets by analyzing shoot structure and volatiles production. In vitro co-cultures were established with the PWN and the effect of the phytoparasite on in vitro shoot structure, water content and volatiles production was evaluated. In vitro shoots showed similar structure and volatiles production to in vivo maritime pine plantlets. The first macroscopic symptoms of PWD were observed about 4 weeks after in vitro co-culture establishment. Nematode population in the culture medium increased and PWNs were detected in gaps of the callus tissue and in cavities developed from the degradation of cambial cells. In terms of volatiles main components, plantlets, P. pinaster cultures, and P. pinaster with B. xylophilus co-cultures were all β- and α-pinene rich. Co-cultures may be an easy-to-handle biotechnological approach to study this pathology, envisioning the understanding of and finding ways to restrain this highly devastating nematode.

Co-culture with Sertoli cells promotes proliferation and migration of umbilical cord mesenchymal stem cells

International Nuclear Information System (INIS)

Zhang, Fenxi; Hong, Yan; Liang, Wenmei; Ren, Tongming; Jing, Suhua; Lin, Juntang

2012-01-01

Highlights: ► Co-culture of Sertoli cells (SCs) with human umbilical cord mesenchymal stem cells (UCMSCs). ► Presence of SCs dramatically increased proliferation and migration of UCMSCs. ► Presence of SCs stimulated expression of Mdm2, Akt, CDC2, Cyclin D, CXCR4, MAPKs. -- Abstract: Human umbilical cord mesenchymal stem cells (hUCMSCs) have been recently used in transplant therapy. The proliferation and migration of MSCs are the determinants of the efficiency of MSC transplant therapy. Sertoli cells are a kind of “nurse” cells that support the development of sperm cells. Recent studies show that Sertoli cells promote proliferation of endothelial cells and neural stem cells in co-culture. We hypothesized that co-culture of UCMSCs with Sertoli cells may also promote proliferation and migration of UCMSCs. To examine this hypothesis, we isolated UCMSCs from human cords and Sertoli cells from mouse testes, and co-cultured them using a Transwell system. We found that UCMSCs exhibited strong proliferation ability and potential to differentiate to other cell lineages such as osteocytes and adipocytes. The presence of Sertoli cells in co-culture significantly enhanced the proliferation and migration potential of UCMSCs (P < 0.01). Moreover, these phenotypic changes were accompanied with upregulation of multiple genes involved in cell proliferation and migration including phospho-Akt, Mdm2, phospho-CDC2, Cyclin D1, Cyclin D3 as well as CXCR4, phospho-p44 MAPK and phospho-p38 MAPK. These findings indicate that Sertoli cells boost UCMSC proliferation and migration potential.

Co-culture with Sertoli cells promotes proliferation and migration of umbilical cord mesenchymal stem cells

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Zhang, Fenxi, E-mail: fxzhang0824@gmail.com [Department of Anatomy, Sanquan College, Xinxiang Medical University, Henan 453003, People' s Republic of China (China); Hong, Yan; Liang, Wenmei [Department of Histology and Embryology, Guiyang Medical University, Guizhou 550004, People' s Republic of China (China); Ren, Tongming [Department of Anatomy, Sanquan College, Xinxiang Medical University, Henan 453003, People' s Republic of China (China); Jing, Suhua [ICU Center, The Third Hospital of Xinxiang Medical University, Henan 453003, People' s Republic of China (China); Lin, Juntang [Stem Cell Center, Xinxiang Medical University, Henan 453003, People' s Republic of China (China)

2012-10-12

Highlights: Black-Right-Pointing-Pointer Co-culture of Sertoli cells (SCs) with human umbilical cord mesenchymal stem cells (UCMSCs). Black-Right-Pointing-Pointer Presence of SCs dramatically increased proliferation and migration of UCMSCs. Black-Right-Pointing-Pointer Presence of SCs stimulated expression of Mdm2, Akt, CDC2, Cyclin D, CXCR4, MAPKs. -- Abstract: Human umbilical cord mesenchymal stem cells (hUCMSCs) have been recently used in transplant therapy. The proliferation and migration of MSCs are the determinants of the efficiency of MSC transplant therapy. Sertoli cells are a kind of 'nurse' cells that support the development of sperm cells. Recent studies show that Sertoli cells promote proliferation of endothelial cells and neural stem cells in co-culture. We hypothesized that co-culture of UCMSCs with Sertoli cells may also promote proliferation and migration of UCMSCs. To examine this hypothesis, we isolated UCMSCs from human cords and Sertoli cells from mouse testes, and co-cultured them using a Transwell system. We found that UCMSCs exhibited strong proliferation ability and potential to differentiate to other cell lineages such as osteocytes and adipocytes. The presence of Sertoli cells in co-culture significantly enhanced the proliferation and migration potential of UCMSCs (P < 0.01). Moreover, these phenotypic changes were accompanied with upregulation of multiple genes involved in cell proliferation and migration including phospho-Akt, Mdm2, phospho-CDC2, Cyclin D1, Cyclin D3 as well as CXCR4, phospho-p44 MAPK and phospho-p38 MAPK. These findings indicate that Sertoli cells boost UCMSC proliferation and migration potential.

Assessment of cellular materials generated by co-cultured ‘inflamed’ and healthy periodontal ligament stem cells from patient-matched groups

Energy Technology Data Exchange (ETDEWEB)

Tang, Hao-Ning [State Key Laboratory of Military Stomatology & National Clinical Research Center for Oral Diseases & Shaanxi Key Laboratory of Stomatology, Department of Periodontology, School of Stomatology, Fourth Military Medical University, Xi' an (China); Department of Stomatology, The First Affiliated Hospital of the Chinese PLA General Hospital, Beijing 100048 (China); Xia, Yu [State Key Laboratory of Military Stomatology & National Clinical Research Center for Oral Diseases & Shaanxi Key Laboratory of Stomatology, Department of Periodontology, School of Stomatology, Fourth Military Medical University, Xi' an (China); Department of Stomatology, The 309th Hospital of Chinese People' s Liberation Army, Beijing 100091 (China); Xu, Jie; Tian, Bei-Min; Zhang, Xi-Yu [State Key Laboratory of Military Stomatology & National Clinical Research Center for Oral Diseases & Shaanxi Key Laboratory of Stomatology, Department of Periodontology, School of Stomatology, Fourth Military Medical University, Xi' an (China); Chen, Fa-Ming, E-mail: cfmsunhh@fmmu.edu.cn [State Key Laboratory of Military Stomatology & National Clinical Research Center for Oral Diseases & Shaanxi Key Laboratory of Stomatology, Department of Periodontology, School of Stomatology, Fourth Military Medical University, Xi' an (China)

2016-08-01

Recently, stem cells derived from the'inflamed’ periodontal ligament (PDL) tissue of periodontally diseased teeth (I-PDLSCs) have been increasingly suggested as a more readily accessible source of cells for regenerative therapies than those derived from healthy PDL tissue (H-PDLSCs). However, substantial evidence indicates that I-PDLSCs exhibit impaired functionalities compared with H-PDLSCs. In this study, patient-matched I-PDLSCs and H-PDLSCs were co-cultured at various ratios. Cellular materials derived from these cultures were investigated regarding their osteogenic potential in vitro and capacity to form new bone following in vivo transplantation. While patient-matched I-PDLSCs and H-PDLSCs could co-exist in co-culture systems, the proportion of I-PDLSCs tended to increase during in vitro incubation. Compared with H-PDLSC monoculture, the presence of I-PDLSCs in the co-cultures appeared to enhance the overall cell proliferation. Although not completely rescued, the osteogenic and regenerative potentials of the cellular materials generated by co-cultured I-PDLSCs and H-PDLSCs were significantly improved compared with those derived from I-PDLSC monocultures. Notably, cells in co-cultures containing either 50% I-PDLSCs plus 50% H-PDLSCs or 25% I-PDLSCs plus 75% H-PDLSCs expressed osteogenesis-related proteins and genes at levels similar to those expressed in H-PDLSC monocultures (P>0.05). Irrespective of the percentage of I-PDLSCs, robust cellular materials were obtained from co-cultures with 50% or more H-PDLSCs, which exhibited equivalent potential to form new bone in vivo compared with sheets generated by H-PDLSC monocultures. These data suggest that the co-culture of I-PDLSCs with patient-matched H-PDLSCs is a practical and effective method for increasing the overall osteogenic and regenerative potentials of resultant cellular materials. - Highlights: • Co-culturing H-PDLSCs with I-PDLSCs led to rapid cell expansion. • H-PDLSCs and I-PDLSCs co-cultured

Assessment of cellular materials generated by co-cultured ‘inflamed’ and healthy periodontal ligament stem cells from patient-matched groups

International Nuclear Information System (INIS)

Tang, Hao-Ning; Xia, Yu; Xu, Jie; Tian, Bei-Min; Zhang, Xi-Yu; Chen, Fa-Ming

2016-01-01

Recently, stem cells derived from the'inflamed’ periodontal ligament (PDL) tissue of periodontally diseased teeth (I-PDLSCs) have been increasingly suggested as a more readily accessible source of cells for regenerative therapies than those derived from healthy PDL tissue (H-PDLSCs). However, substantial evidence indicates that I-PDLSCs exhibit impaired functionalities compared with H-PDLSCs. In this study, patient-matched I-PDLSCs and H-PDLSCs were co-cultured at various ratios. Cellular materials derived from these cultures were investigated regarding their osteogenic potential in vitro and capacity to form new bone following in vivo transplantation. While patient-matched I-PDLSCs and H-PDLSCs could co-exist in co-culture systems, the proportion of I-PDLSCs tended to increase during in vitro incubation. Compared with H-PDLSC monoculture, the presence of I-PDLSCs in the co-cultures appeared to enhance the overall cell proliferation. Although not completely rescued, the osteogenic and regenerative potentials of the cellular materials generated by co-cultured I-PDLSCs and H-PDLSCs were significantly improved compared with those derived from I-PDLSC monocultures. Notably, cells in co-cultures containing either 50% I-PDLSCs plus 50% H-PDLSCs or 25% I-PDLSCs plus 75% H-PDLSCs expressed osteogenesis-related proteins and genes at levels similar to those expressed in H-PDLSC monocultures (P>0.05). Irrespective of the percentage of I-PDLSCs, robust cellular materials were obtained from co-cultures with 50% or more H-PDLSCs, which exhibited equivalent potential to form new bone in vivo compared with sheets generated by H-PDLSC monocultures. These data suggest that the co-culture of I-PDLSCs with patient-matched H-PDLSCs is a practical and effective method for increasing the overall osteogenic and regenerative potentials of resultant cellular materials. - Highlights: • Co-culturing H-PDLSCs with I-PDLSCs led to rapid cell expansion. • H-PDLSCs and I-PDLSCs co-cultured

Transdifferentiation of mouse adipose-derived stromal cells into acinar cells of the submandibular gland using a co-culture system

International Nuclear Information System (INIS)

Lee, Jingu; Park, Sangkyu; Roh, Sangho

2015-01-01

A loss of salivary gland function often occurs after radiation therapy in head and neck tumors, though secretion of saliva by the salivary glands is essential for the health and maintenance of the oral environment. Transplantation of salivary acinar cells (ACs), in part, may overcome the side effects of therapy. Here we directly differentiated mouse adipose-derived stromal cells (ADSCs) into ACs using a co-culture system. Multipotent ADSCs can be easily collected from stromal vascular fractions of adipose tissues. The isolated ADSCs showed positive expression of markers such as integrin beta-1 (CD29), cell surface glycoprotein (CD44), endoglin (CD105), and Nanog. The cells were able to differentiate into adipocytes, osteoblasts, and neural-like cells after 14 days in culture. ADSCs at passage 2 were co-cultured with mouse ACs in AC culture medium using the double-chamber (co-culture system) to avoid mixing the cell types. The ADSCs in this co-culture system expressed markers of ACs, such as α-amylases and aquaporin5, in both mRNA and protein. ADSCs cultured in AC-conditioned medium also expressed AC markers. Cellular proliferation and senescence analyses demonstrated that cells in the co-culture group showed lower senescence and a higher proliferation rate than the AC-conditioned medium group at Days 14 and 21. The results above imply direct conversion of ADSCs into ACs under the co-culture system; therefore, ADSCs may be a stem cell source for the therapy for salivary gland damage. - Highlights: • ADSCs could transdifferentiate into acinar cells (ACs) using ACs co-culture (CCA). • Transdifferentiated ADSCs expressed ACs markers such as α-amylase and aquaporin5. • High proliferation and low senescence were presented in CCA at Day 14. • Transdifferentiation of ADSCs into ACs using CCA may be an appropriate method for cell-based therapy

Transdifferentiation of mouse adipose-derived stromal cells into acinar cells of the submandibular gland using a co-culture system

Energy Technology Data Exchange (ETDEWEB)

Lee, Jingu; Park, Sangkyu; Roh, Sangho, E-mail: sangho@snu.ac.kr

2015-05-15

A loss of salivary gland function often occurs after radiation therapy in head and neck tumors, though secretion of saliva by the salivary glands is essential for the health and maintenance of the oral environment. Transplantation of salivary acinar cells (ACs), in part, may overcome the side effects of therapy. Here we directly differentiated mouse adipose-derived stromal cells (ADSCs) into ACs using a co-culture system. Multipotent ADSCs can be easily collected from stromal vascular fractions of adipose tissues. The isolated ADSCs showed positive expression of markers such as integrin beta-1 (CD29), cell surface glycoprotein (CD44), endoglin (CD105), and Nanog. The cells were able to differentiate into adipocytes, osteoblasts, and neural-like cells after 14 days in culture. ADSCs at passage 2 were co-cultured with mouse ACs in AC culture medium using the double-chamber (co-culture system) to avoid mixing the cell types. The ADSCs in this co-culture system expressed markers of ACs, such as α-amylases and aquaporin5, in both mRNA and protein. ADSCs cultured in AC-conditioned medium also expressed AC markers. Cellular proliferation and senescence analyses demonstrated that cells in the co-culture group showed lower senescence and a higher proliferation rate than the AC-conditioned medium group at Days 14 and 21. The results above imply direct conversion of ADSCs into ACs under the co-culture system; therefore, ADSCs may be a stem cell source for the therapy for salivary gland damage. - Highlights: • ADSCs could transdifferentiate into acinar cells (ACs) using ACs co-culture (CCA). • Transdifferentiated ADSCs expressed ACs markers such as α-amylase and aquaporin5. • High proliferation and low senescence were presented in CCA at Day 14. • Transdifferentiation of ADSCs into ACs using CCA may be an appropriate method for cell-based therapy.

Human breast cancer histoid: an in vitro 3-dimensional co-culture model that mimics breast cancer tissue.

Science.gov (United States)

Kaur, Pavinder; Ward, Brenda; Saha, Baisakhi; Young, Lillian; Groshen, Susan; Techy, Geza; Lu, Yani; Atkinson, Roscoe; Taylor, Clive R; Ingram, Marylou; Imam, S Ashraf

2011-12-01

Progress in our understanding of heterotypic cellular interaction in the tumor microenvironment, which is recognized to play major roles in cancer progression, has been hampered due to unavailability of an appropriate in vitro co-culture model. The aim of this study was to generate an in vitro 3-dimensional human breast cancer model, which consists of cancer cells and fibroblasts. Breast cancer cells (UACC-893) and fibroblasts at various densities were co-cultured in a rotating suspension culture system to establish co-culture parameters. Subsequently, UACC-893, BT.20, or MDA.MB.453 were co-cultured with fibroblasts for 9 days. Co-cultures resulted in the generation of breast cancer histoid (BCH) with cancer cells showing the invasion of fibroblast spheroids, which were visualized by immunohistochemical (IHC) staining of sections (4 µm thick) of BCH. A reproducible quantitative expression of C-erbB.2 was detected in UACC-893 cancer cells in BCH sections by IHC staining and the Automated Cellular Imaging System. BCH sections also consistently exhibited qualitative expression of pancytokeratins, p53, Ki-67, or E-cadherin in cancer cells and that of vimentin or GSTPi in fibroblasts, fibronectin in the basement membrane and collagen IV in the extracellular matrix. The expression of the protein analytes and cellular architecture of BCH were markedly similar to those of breast cancer tissue.

Allogeneic human dermal fibroblasts are viable in peripheral blood mononuclear co-culture

Directory of Open Access Journals (Sweden)

Restu Syamsul Hadi

2014-08-01

Full Text Available Background Transplanted allogeneic dermal fibroblasts retain stem cell subpopulations, and are easily isolated, expanded and stored using standard techniques. Their potential for regenerative therapy of chronic wounds should be evaluated. The aim of this study was to determine allogeneic fibroblast viability in the presence of peripheral blood mononuclear cells (PBMC. Methods In this experimental study, fibroblasts were isolated from foreskin explants, expanded in the presence of serum, and stored using slow-freezing. We used one intervention group of allogeneic fibroblasts co-cultured with PBMC and 2 control groups of separate fibroblast and PBMC cultures.Fibroblasts were characterized by their collagen secretion and octamer-binding transcription factor 4 (OCT4 expression. Viability was evaluated using water soluble tetrazolium-1 (WST-1 proliferation assay. Absorbances were measured at 450 nm. Data analysis was performed by student’s paired t-test. Results Dermal fibroblasts were shown to secrete collagen, express OCT4, be recoverable after cryopreservation, and become attached to the culture dish in a co-culture with PBMC. Co-cultured and control fibroblasts had no significantly different cell viabilities (p>0.05. Calculated viable cell numbers increased 1.8 and 5.1-fold, respectively, at days 2 and 4 in vitro. Both groups showed comparable doubling times at days 2 and 4 in vitro. PBMC did not interfere with allogeneic fibroblast viability and proliferative capacity Conclusions Allogeneic fibroblasts remain viable and proliferate in the presence of host PBMC. Future research should evaluate allogeneic human dermal fibroblast competency in clinical settings. Dermal fibroblasts are a potential source for cell therapy in chronic wound management.

3D cellular structures and co-cultures formed through the contactless magnetic manipulation of cells on adherent surfaces.

Science.gov (United States)

Abdel Fattah, Abdel Rahman; Mishriki, Sarah; Kammann, Tobias; Sahu, Rakesh P; Geng, Fei; Puri, Ishwar K

2018-02-27

A magnet array is employed to manipulate diamagnetic cells that are contained in paramagnetic medium to demonstrate for the first time the contactless bioprinting of three-dimensional (3D) cellular structures and co-cultures of breast cancer MCF-7 and endothelial HUVEC at prescribed locations on tissue culture treated well plates. Sequential seeding of different cell lines and the spatial displacement of the magnet array creates co-cultured cellular structures within a well without using physically intrusive well inserts. Both monotypic and co-culture experiments produce morphologically rich 3D cell structures that are otherwise absent in regular monolayer cell cultures. The magnetic contactless bioprinting of cells provides further insight into cell behaviour, invasion strategies and transformations that are useful for potential applications in drug screening, 3D cell culture formation and tissue engineering.

Myofibroblast androgen receptor expression determines cell survival in co-cultures of myofibroblasts and prostate cancer cells in vitro.

Science.gov (United States)

Palethorpe, Helen M; Leach, Damien A; Need, Eleanor F; Drew, Paul A; Smith, Eric

2018-04-10

Fibroblasts express androgen receptor (AR) in the normal prostate and during prostate cancer development. We have reported that loss of AR expression in prostate cancer-associated fibroblasts is a poor prognostic indicator. Here we report outcomes of direct and indirect co-cultures of immortalised AR-positive (PShTert-AR) or AR-negative (PShTert) myofibroblasts with prostate cancer cells. In the initial co-cultures the AR-negative PC3 cell line was used so AR expression and signalling were restricted to the myofibroblasts. In both direct and indirect co-culture with PShTert-AR myofibroblasts, paracrine signalling to the PC3 cells slowed proliferation and induced apoptosis. In contrast, PC3 cells proliferated with PShTert myofibroblasts irrespective of the co-culture method. In direct co-culture PC3 cells induced apoptosis in and destroyed PShTerts by direct signalling. Similar results were seen in direct co-cultures with AR-negative DU145 and AR-positive LNCaP and C4-2B prostate cancer cell lines. The AR ligand 5α-dihydrotestosterone (DHT) inhibited the proliferation of the PShTert-AR myofibroblasts, thereby reducing the extent of their inhibitory effect on cancer cell growth. These results suggest loss of stromal AR would favour prostate cancer cell growth in vivo , providing an explanation for the clinical observation that reduced stromal AR is associated with a poorer outcome.

[Experimental study on co-culture of human fibroblasts on decellularized Achilles tendon].

Science.gov (United States)

Wang, Zhibing; Zhang, Xia; Guo, Xinyu; Qin, Chuan

2013-07-01

To investigate the preparation of decellularized Achilles tendons and the effect of co-culture of human fibroblasts on the scaffold so as to provide a scaffold for the tissue engineered ligament reconstruction. Achilles tendons of both hind limbs were harvested from 10 male New Zealand white rabbits (5-month-old; weighing, 4-5 kg). The Achilles tendons were decellularized using trypsin, Triton X-100, and sodium dodecyl sulfate (SDS), and then gross observation, histological examination, and scanning electron microscope (SEM) observation were performed; the human fibroblasts were seeded on the decellularized Achilles tendon, and then cytocompatibility was tested using the cell counting kit 8 method at 1, 3, 5, 7, and 9 days after co-culture. At 4 weeks after co-culture, SEM, HE staining, and biomechanical test were performed for observing cell-scaffold composite, and a comparison was made with before and after decellularization. After decellularization, the tendons had integrated aponeurosis and enlarged volume with soft texture and good toughness; there was no loose connective tissue and tendon cells between tendon bundles, the collagen fibers arranged loosely with three-dimensional network structure and more pores between tendon bundles; and it had good cytocompatibility. At 4 weeks after co-culture, cells migrated into the pores, and three-dimensional network structure disappeared. By biomechanical test, the tensile strength and Young's elastic modulus of the decellularized Achilles tendon group decreased significantly when compared with normal Achilles tendons group and cell-scaffold composite group (P Achilles tendons group and cell-scaffold composite group (P > 0.05). There was no significant difference in elongation at break among 3 groups (P > 0.05). The decellularized Achilles tendon is biocompatible to fibroblasts. It is suit for the scaffold for tissue engineered ligament reconstruction.

Quorum sensing signals are produced by Aeromonas salmonicida and quorum sensing inhibitors can reduce production of a potential virulence factor

DEFF Research Database (Denmark)

Rasch, Maria; Kastbjerg, Vicky Gaedt; Bruhn, Jesper Bartholin

2007-01-01

Many pathogens control production of virulence factors by self-produced signals in a process called quorum sensing (QS). We demonstrate that acyl homoserine lactone (AHL) signals, which enable bacteria to express certain phenotypes in relation to cell density, are produced by a wide spectrum...... of Aeromonas salmonicida strains. All 31 typical strains were AHL producers as were 21 of 26 atypical strains, but on a strain population basis, production of virulence factors such as protease, lipase, A-layer or pigment did not correlate with the production and accumulation of AHLs in the growth medium...... of Aeromonas salmonicida. The most efficient compound N-(heptylsulfanylacetyl)-L-homoserine lactone (HepS-AHL), reduced protease production by a factor of 10. Five extracellular proteases were detected on gelatin-containing sodium dodecyl sulphate polyacrylamide gel electrophoresis (SDS-PAGE) gels and 3...

Positive correlation between Aeromonas salmonicida vaccine antigen concentration and protection in vaccinated rainbow trout Oncorhynchus mykiss evaluated by a tail fin infection model

DEFF Research Database (Denmark)

Marana, M. H.; Skov, J.; Chettri, Jiwan Kumar

2017-01-01

Rainbow trout, Oncorhynchus mykiss (Walbaum), are able to raise a protective immune response against Aeromonas salmonicida subsp. salmonicida (AS) following injection vaccination with commercial vaccines containing formalin-killed bacteria, but the protection is often suboptimal under Danish...... mariculture conditions. We elucidated whether protection can be improved by increasing the concentration of antigen (formalin-killed bacteria) in the vaccine. Rainbow trout juveniles were vaccinated by intraperitoneal (i.p.) injection with a bacterin of Aeromonas salmonicida subsp. salmonicida strain 090710...... bacteria. The infection method proved to be efficient and could differentiate efficacies of different vaccines. It was shown that protection and antibody production in exposed fish were positively correlated to the AS antigen concentration in the vaccine....

Combination of Aloe vera and xenograft induction on decreasing of NF-kb of tooth extraction socket preservation in Cavia cobaya

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Utari Kresnoadi

2014-03-01

Full Text Available Background: Tooth extraction can naturally cause inflammation triggering osteoclast proliferation and alveolar bone resorption. Preservation of the tooth extraction sockets is needed for patients in order to reduce alveolar bone resorption risks. Aloe vera is known to have anthraquinones components, namely Aloin, Aloe emedin, and barbaloin, considered as anti-inflammation. Therefore, to overcome the inflammation, the role of NF-kb is very significant to decrease nuclear factor kappa b (NF-kb. As a result, inflammation risks will be decreased. Purpose: The study was aimed to determine the induction effect of combination of Aloe vera and XCB into tooth extraction sockets to reduce inflammation by reducing NF-kb expression, osteoclasts and osteoblasts. Methods: Forty-eight Cavia cobaya were divided into eight groups, each group consisted of six animals. The mandibular incisors of those Cavia cobaya were extracted and induced with either PEG, XCB, Aloe vera, or the combination of Aloe vera + XCB. Those animals were sacrificed on day 7 and day 30 after the extraction. Then immunohistochemical and histopathology examinations were conducted to observe NF-kb expression, osteoblasts and osteoclasts. Results: It was known that in group induced with the combination of Aloe vera and xenograft concelous bovine, the growth of osteoblasts was high, while NF-kb expression and osteoclasts reduced. Conclusion: It can be concluded that the induction of the combination of Aloe vera and XCB into the tooth extraction sockets can reduce NF-kb expression and osteoclast, as a result, alveolar bone resorption risks decrease, and osteoblast increase.Latar belakang: Trauma mekanis akibat pencabutan gigi asli menyebabkan keradangan. Keradangan memicu proliferasi osteoklas sehingga menyebabkan resorpsi tulang alveolararis. Pada pembuatan gigi tiruan, resorpsi tulang alveolar yang terjadi, sangat tidak diinginkan, sebab resorpsi tulang alveolar mengurangi keberhasilan

Conseil de la Cooperation Culturelle et Fonds Culturel. Rapport Annuel 1972. (Council for Cultural Co-operation and Cultural Fund. Annual Report 1972.)

Science.gov (United States)

Council for Cultural Cooperation, Strasbourg (France).

This document is a report on the activities of the Council for Cultural Co-Operation (CCC) and Cultural Fund for 1972. The CCC is briefly defined as being designed to reinforce educational and cultural cooperation among the 21 countries of the CCC, especially in matters of research, information, and documentation. The document is composed of…

A DNA biosensor for molecular diagnosis of Aeromonas hydrophila using zinc sulfide nanospheres

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M. Negahdary

2017-07-01

Full Text Available Today, identification of pathogenic bacteria using modern and accurate methods is inevitable. Integration in electrochemical measurements with nanotechnology has led to the design of efficient and sensitive DNA biosensors against bacterial agents. Here, efforts were made to detect Aeromonas hydrophila using aptamers as probes and zinc sulfide (ZnS nanospheres as signal enhancers and electron transfer facilitators. After modification of the working electrode area (in a screen-printed electrode with ZnS nanospheres through electrodeposition, the coated surface of a modified electrode with ZnS nanospheres was investigated through scanning electron microscopy (SEM. The size of synthesized ZnS nanospheres was estimated at about 20–50 nm and their shape was in the form of porous plates in microscopic observations. All electrochemical measurements were performed using cyclic voltammetry (CV, electrochemical impedance spectroscopy (EIS, and constant potential amperometry (CPA techniques. The designed DNA biosensor was able to detect deoxyribonucleic acid (DNA of Aeromonas hydrophila in the range 1.0  ×  10−4 to 1.0  ×  10−9 mol L−1; the limit of detection (LOD in this study was 1  ×  10−13 mol L−1. This DNA biosensor showed satisfactory thermal and pH stability. Reproducibility for this DNA biosensor was measured and the relative standard deviation (RSD of the performance of this DNA biosensor was calculated as 5 % during 42 days.

Cultures and co-cultures of human blood mononuclear cells and endothelial cells for the biocompatibility assessment of surface modified AISI 316L austenitic stainless steel

Energy Technology Data Exchange (ETDEWEB)

Stio, Maria; Martinesi, Maria; Treves, Cristina [Dipartimento di Scienze Biomediche, Sperimentali e Cliniche ‘Mario Serio’, Sezione di Scienze Biochimiche, Università di Firenze, viale Morgagni 50, 50134 Firenze (Italy); Borgioli, Francesca, E-mail: francesca.borgioli@unifi.it [Dipartimento di Ingegneria Industriale (DIEF), Università di Firenze, via S. Marta 3, 50139 Firenze (Italy)

2016-12-01

Samples of AISI 316L austenitic stainless steel were subjected either to grinding and polishing procedure, or to grinding and then low temperature glow-discharge nitriding treatment, or to grinding, nitriding and subsequently coating with collagen-I. Nitrided samples, even if only ground, show a higher corrosion resistance in PBS solution, in comparison with ground and polished AISI 316L. Biocompatibility was evaluated in vitro by incubating the samples with either peripheral blood mononuclear cells (PBMC) or human umbilical vein endothelial cells (HUVEC), tested separately or in co-culture. HUVEC-PBMC co-culture and co-incubation of HUVEC with PBMC culture medium, after the previous incubation of PBMC with metallic samples, allowed to determine whether the incubation of PBMC with the different samples might affect HUVEC behaviour. Many biological parameters were considered: cell proliferation, release of cytokines, matrix metalloproteinases (MMPs) and sICAM-1, gelatinolytic activity of MMPs, and ICAM-1 protein expression. Nitriding treatment, with or without collagen coating of the samples, is able to ameliorate some of the biological parameters taken into account. The obtained results point out that biocompatibility may be successfully tested in vitro, using cultures of normal human cells, as blood and endothelial cells, but more than one cell line should be used, separately or in co-culture, and different parameters should be determined, in particular those correlated with inflammatory phenomena. - Highlights: • Nitriding improves corrosion resistance and biocompatibility of ground AISI 316L. • The metallic samples differently affect different human cell cultures. • PBMC and HUVEC are a suitable model to test in vitro biocompatibility. • Co-cultures show that HUVEC are affected by pre-incubation of PBMC with the samples. • Inflammation parameters must be taken into account for assessing biocompatibility.

Cultures and co-cultures of human blood mononuclear cells and endothelial cells for the biocompatibility assessment of surface modified AISI 316L austenitic stainless steel

International Nuclear Information System (INIS)

Stio, Maria; Martinesi, Maria; Treves, Cristina; Borgioli, Francesca

2016-01-01

Samples of AISI 316L austenitic stainless steel were subjected either to grinding and polishing procedure, or to grinding and then low temperature glow-discharge nitriding treatment, or to grinding, nitriding and subsequently coating with collagen-I. Nitrided samples, even if only ground, show a higher corrosion resistance in PBS solution, in comparison with ground and polished AISI 316L. Biocompatibility was evaluated in vitro by incubating the samples with either peripheral blood mononuclear cells (PBMC) or human umbilical vein endothelial cells (HUVEC), tested separately or in co-culture. HUVEC-PBMC co-culture and co-incubation of HUVEC with PBMC culture medium, after the previous incubation of PBMC with metallic samples, allowed to determine whether the incubation of PBMC with the different samples might affect HUVEC behaviour. Many biological parameters were considered: cell proliferation, release of cytokines, matrix metalloproteinases (MMPs) and sICAM-1, gelatinolytic activity of MMPs, and ICAM-1 protein expression. Nitriding treatment, with or without collagen coating of the samples, is able to ameliorate some of the biological parameters taken into account. The obtained results point out that biocompatibility may be successfully tested in vitro, using cultures of normal human cells, as blood and endothelial cells, but more than one cell line should be used, separately or in co-culture, and different parameters should be determined, in particular those correlated with inflammatory phenomena. - Highlights: • Nitriding improves corrosion resistance and biocompatibility of ground AISI 316L. • The metallic samples differently affect different human cell cultures. • PBMC and HUVEC are a suitable model to test in vitro biocompatibility. • Co-cultures show that HUVEC are affected by pre-incubation of PBMC with the samples. • Inflammation parameters must be taken into account for assessing biocompatibility.

Microbial reductive dehalogenation of trihalomethanes by a Dehalobacter-containing co-culture.

Science.gov (United States)

Zhao, Siyan; Rogers, Matthew J; He, Jianzhong

2017-07-01

Trihalomethanes such as chloroform and bromoform, although well-known as a prominent class of disinfection by-products, are ubiquitously distributed in the environment due to widespread industrial usage in the past decades. Chloroform and bromoform are particularly concerning, of high concentrations detected and with long half-lives up to several hundred days in soils and groundwater. In this study, we report a Dehalobacter- and Desulfovibrio-containing co-culture that exhibits dehalogenation of chloroform (~0.61 mM) to dichloromethane and bromoform (~0.67 mM) to dibromomethane within 10-15 days. This co-culture was further found to dechlorinate 1,1,1-trichloroethane (1,1,1-TCA) (~0.65 mM) to 1,1-dichloroethane within 12 days. The Dehalobacter species present in this co-culture, designated Dehalobacter sp. THM1, was found to couple growth with dehalogenation of chloroform, bromoform, and 1,1,1-TCA. Strain THM1 harbors a newly identified reductive dehalogenase (RDase), ThmA, which catalyzes chloroform, bromoform, and 1,1,1-TCA dehalogenation. Additionally, based on the sequences of thmA and other identified chloroform RDase genes, ctrA, cfrA, and tmrA, a pair of chloroform RDase gene-specific primers were designed and successfully applied to investigate the chloroform dechlorinating potential of microbial communities. The comparative analysis of chloroform RDases with tetrachloroethene RDases suggests a possible approach in predicting the substrate specificity of uncharacterized RDases in the future.

sugE: A gene involved in tributyltin (TBT) resistance of Aeromonas molluscorum Av27.

Science.gov (United States)

Cruz, Andreia; Micaelo, Nuno; Félix, Vitor; Song, Jun-Young; Kitamura, Shin-Ichi; Suzuki, Satoru; Mendo, Sónia

2013-01-01

The mechanism of bacterial resistance to tributyltin (TBT) is still unclear. The results herein presented contribute to clarify that mechanism in the TBT-resistant bacterium Aeromonas molluscorum Av27. We have identified and cloned a new gene that is involved in TBT resistance in this strain. The gene is highly homologous (84%) to the Aeromonas hydrophila-sugE gene belonging to the small multidrug resistance gene family (SMR), which includes genes involved in the transport of lipophilic drugs. In Av27, expression of the Av27-sugE was observed at the early logarithmic growth phase in the presence of a high TBT concentration (500 μM), thus suggesting the contribution of this gene for TBT resistance. E. coli cells transformed with Av27-sugE become resistant to ethidium bromide (EtBr), chloramphenicol (CP) and tetracycline (TE), besides TBT. According to the Moriguchi logP (miLogP) values, EtBr, CP and TE have similar properties and are substrates for the sugE-efflux system. Despite the different miLogP of TBT, E. coli cells transformed with Av27-sugE become resistant to this compound. So it seems that TBT is also a substrate for the SugE protein. The modelling studies performed also support this hypothesis. The data herein presented clearly indicate that sugE is involved in TBT resistance of this bacterium.

Antibiotic resistence of Aeromonas hydrophila isolated from Piaractus mesopotamicus (Holmberg, 1887) and Oreochromis niloticus (Linnaeus, 1758)

OpenAIRE

Belém-Costa,Andréa; Cyrino,José Eurico Possebon

2006-01-01

One of the most important problems involving treatments with antibiotics against Aeromonas hydrophila isolated from fishes is that antibiotic resistance develops readily. The antimicrobial activity of chemotherapeutants in isolates from pacu Piaractus mesopotamicus (Holmberg, 1887) and tilapia Oreochromis niloticus (Linnaeus, 1758) was tested by the Kirby-Bauer disk method, over Mueller-Hinton surface agar previously inoculated with 100 µL of bacterial suspensions. After regular incubation, i...

Ethanol production from corn cobs by co-culture of Saccharomyces ...

African Journals Online (AJOL)

Saccharomyces cerevisiae and Aspergillus niger were used in a co-culture for the simultaneous saccharification and fermentation (SSF) of 1% and 10% (w/v) dry pre-treated corn cobs to ethanol. Positive controls of glucose of same concentrations in a synthetic medium were also fermented. At 1% substrate concentration, ...

Temperature Preference in IAF Hairless and Hartley Guinea Pigs (Cavia porcellus).

Science.gov (United States)

Kleven, Gale A; Joshi, Prianca

2016-03-01

The Hairless strain of guinea pigs (Cavia porcellus) is the result of a spontaneous recessive mutation first identified at the Institute Armand Frappier (IAF) in 1978. Despite the longstanding availability of this strain, little is known about its thermoregulatory behavior. The aim of this study was to determine temperature preference in Hartley and Hairless guinea pigs by observing each strain in a ring-shaped apparatus containing a nonlinear temperature gradient. Temperatures were maintained by separately controlled heating mats lining the apparatus. Set point temperatures ranged from 24 to 38 °C. Guinea pigs (Hartley female, Hairless female, and Hairless male guinea pigs; n = 8 each group) were placed either singly or in pairs at 1 of the 8 randomized starting points within the apparatus. Subjects were observed for 30 min and coded for location within the temperature gradient by both frequency and duration. When placed singly in the apparatus, all 3 groups spent more time in the 30 °C zones. However, when placed as pairs with a cagemate, Hartley female guinea pigs spent more time in the cooler range of temperatures from 24 to 30 °C, whereas Hairless guinea pigs preferred a range of 30 to 38 °C. These results confirm a temperature preference of 30 ± 2 °C for both Hartley and Hairless guinea pigs when singly housed. However, data from the paired housing condition suggest that context plays an important role in thermoregulatory behavior.

Discovery of novel xylosides in co-culture of basidiomycetes Trametes versicolor and Ganoderma applanatum by integrated metabolomics and bioinformatics

Science.gov (United States)

Yao, Lu; Zhu, Li-Ping; Xu, Xiao-Yan; Tan, Ling-Ling; Sadilek, Martin; Fan, Huan; Hu, Bo; Shen, Xiao-Ting; Yang, Jie; Qiao, Bin; Yang, Song

2016-09-01

Transcriptomic analysis of cultured fungi suggests that many genes for secondary metabolite synthesis are presumably silent under standard laboratory condition. In order to investigate the expression of silent genes in symbiotic systems, 136 fungi-fungi symbiotic systems were built up by co-culturing seventeen basidiomycetes, among which the co-culture of Trametes versicolor and Ganoderma applanatum demonstrated the strongest coloration of confrontation zones. Metabolomics study of this co-culture discovered that sixty-two features were either newly synthesized or highly produced in the co-culture compared with individual cultures. Molecular network analysis highlighted a subnetwork including two novel xylosides (compounds 2 and 3). Compound 2 was further identified as N-(4-methoxyphenyl)formamide 2-O-β-D-xyloside and was revealed to have the potential to enhance the cell viability of human immortalized bronchial epithelial cell line of Beas-2B. Moreover, bioinformatics and transcriptional analysis of T. versicolor revealed a potential candidate gene (GI: 636605689) encoding xylosyltransferases for xylosylation. Additionally, 3-phenyllactic acid and orsellinic acid were detected for the first time in G. applanatum, which may be ascribed to response against T.versicolor stress. In general, the described co-culture platform provides a powerful tool to discover novel metabolites and help gain insights into the mechanism of silent gene activation in fungal defense.

Prevalence and diversity of Aeromonas and Vibrio spp. in coastal waters of Southern Italy

DEFF Research Database (Denmark)

Dumontet, S.; Krovacek, K.; Svenson, S.B.

2000-01-01

% of samples were positive for Vibrio spp. It was interesting to note that 38% of the positive stations for both Aeromonas and Vibrio spp. showed a fecal coliform contamination of water at ... coliforms) do not always satisfactorily reflect the hygienic quality of water. The presence of Vibrionaceae on copepods was also investigated. Copepods were sampled at a station located inside the harbour of the city of Naples and were found contaminated by V. cholerae non-Ol, V. alginolyticus, V. fluvialis...

DIFFERENTIATION OF AEROMONAS ISOLATES OBTAINED FROM DRINKING WATER DISTRIBUTION SYSTEM USING MATRIX-ASSISTED LASER DESCRIPTION/IONIZATION-MASS SPECTROMETRY (MALDI-MS)

Science.gov (United States)

The genus Aeromonas is one of several medically significant genera that have gained prominence due to their evolving taxonomy and controversial role in human diseases. In this study, matrix-assisted laser desorption/ionization-mass spectrometry (MALDI-MS) was used to analyze the...

Biosynthesis of phenolic compounds inVitis vinifera cell suspension cultures: Study on hydroxycinnamoyl CoA:ligase.

Science.gov (United States)

Lotfy, S; Lofty, S; Fleuriet, A; Ramos, T; Macheix, J J

1989-02-01

In cell suspensions cultures from grape berry pulp (Vitis vinifera cv. Gamay fréaux)hydroxycinnamoyl CoA ligase (CoAL) displayed maximum activity (100 %) forp-coumaric acid and then, in decreasing order, for ferulic acid (81.3 %) and caffeic acid (60.4 %). No activity was detected with sinapic and cinnamic acids. The changes in CoAL activity during the growth cycle of the culture displayed two peaks : the highest (6 h after subculturing) was linked with a strong increase in protein caused by dilution ; the second was weaker and occurred on the 7th day of culture.Grape cell suspension accumulated mainly peonidin (Pn) and cyanidin (Cy) glucosides (Pn 3-glucoside, Cy 3-glucoside, Pn 3-acetylglucoside, Pn 3-caffeylglucoside, Pn 3-p-coumarylglucoside, and Cy 3-p-coumarylglucoside). Maximum accumulation of anthocyanins was associated with the exponential growth phase of the culture and might be the result of the substantial increase in CoAL activity resulting from the effect of dilution. The second enzyme activity peak was probably oriented towards the acylation of anthocyanins since the percentage of acylated forms increased with time after subculturing.

Identifikasi Gen Aerolysin dan Sensitivitas Antibiotik Aeromonas Hydrophila Penyebab Kematian Tukik (Lepidochelys olivacea di Pulau Serangan, Bali

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Rima Ratnanggana Prasetya

2017-06-01

Full Text Available The purpose of this study was to evaluate the pathogene of Aeromonas hydrophila genes (Aerolysin as the cause of Lepidochelys olivacea death and to perform the antibiotic sensitivity test for antibiotic that often used in order to provide the best antibiotic treatment in the field case. The method used was the identification test using primary cultures on Tryptic Soy Agar (TSA and Mac Conkey Agar (MCA medium, then followed by Gram staining, oxidase, catalase, and biochemical tests. Then, aerolysin genes detection was performed by using Polymerase Chain Reaction (PCR method, while oxytetracycline, chloramphenicol, streptomycin, and kanamycin were tested for their sensitivity. From five samples of dead Lepidochelys olivacea, bleeding on the liver and intestine were observed. The identification result of bacteria was found A. hydrophila, and identified the gene encoding aerolysin. The antibiotic sensitivity test showed a sensitive category of A. hydrophila with a sensitivity level were oxytetracycline, chloramphenicol, kanamycin, and streptomycin. The main cause of Lepidochelys olivacea death in Turtle Conservation and Education Centre, Serangan Island, Bali was A. hydrophila which contains aerolysin genes, in which oxytetracycline seems to be the most effective drug for the treatment. ABSTRAK Tujuan dari penelitian ini adalah membuktikan adanya gen patogen (Aerolysin pada Aeromonas hydrophila sebagai penyebab kematian tukik penyu abu-abu serta mengetahui kepekaan obat antibiotic yang sering digunakan sehingga dapat memberikan terapi yang tepat pada kasus di lapangan. Metode penelitian yang digunakan adalah uji identifikasi menggunakan kultur primer pada media Tryptic Soy Agar (TSA dan Mac Conkey Agar (MCA, dilanjutkan uji pewarnaan Gram, uji oksidase, uji katalase, dan uji biokimiawi. Setelah itu dilakukan deteksi gen aerolysin menggunakan metode Polymerase Chain Reaction (PCR, serta dilakukan uji sensitivitas antibiotik oxytetracycline

Vibrio cholerae No O1 en muestras de aguas no cloradas consumidas por pobladores de las localidades de Santa y Coishco (Ancash, 2003 - 2004

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Ana García P

2006-07-01

Full Text Available Objetivo: Identificar la presencia de Vibrio cholerae en muestras de agua no cloradas para consumo humano en las localidades de Santa y Coishco. Materiales y métodos: Entre julio de 2003 a junio de 2004 se tomaron muestras de agua, en forma semanal, provenientes de siete pozos con bomba manuable y de seis pozos con reservorio. A cada muestra de agua se le midió in situ el cloro residual mediante un comparador de cloro Hatch, método colorimétrico, usando para ello las pastillas DPD 1. En las muestras con cloro <0,05mg/L se realizó el cultivo según los manuales de procedimientos del Instituto Nacional de Salud (INS, Lima. Las cepas aisladas se enviaron al INS para confirmación diagnóstica y pruebas serológicas. Resultados: Se incluyeron 308 muestras de agua para consumo humano en ambos distritos (201 de pozos con bomba manuable y 107 con reservorio. Se realizó el aislamiento en 70(22,7% muestras: Aeromonas caviae 34(11,0%, Aeromonas hidrophyla 17(5,5% y Vibrio cholerae No O1 19(6,2%, no se encontró V. cholerae del serotipo O139. El Vibrio cholerae No O1 se aisló en 11(5,5% muestras de pozos con bomba manuable y en 8(7,4% pozos con reservorio, respectivamente. Conclusión: El agua de consumo humano proveniente de pozos tubulares representa un reservorio potencial para bacterias como Aeromonas y Vibrio cholerae, resaltando la necesidad de realizar la desinfección correspondiente de ésta antes de su consumo.

PENGGUNAAN EKSTRAK DAUN SIRIH (Piper betle Linn UNTUK MENGHAMBAT BAKTERI Aeromonas hydrophila DAN TOKSISITASNYA PADA IKAN PATIN (Pangasius hypophthalmus (THE USE OF PIPER BETLE LINN EXTRACT ON AEROMONAS HYDROPHILA TO OBSTRUCT AND THE TOKSISITY TO PANGASIUS HYPOPHTHALMUS

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Siti Aisiah

2016-06-01

Full Text Available Tujuan penelitian ini untuk mengetahui   daya  hambat  daun  sirih   yang  paling  besar  terhadap  bakteri Aeromonas Hydrophila, mengetahui konsentrasi minimal ekstrak daun sirih yang mampu menghambat pertumbuhan bakteri  A. hydrophila  dan mengetahui toksisitas konsentrasi efektif dari  ekstrak daun sirih terhadap ikan patin. Rancangan percobaan yang digunakan untuk  uji toksisitas  adalah rancangan acak lengkap, terdiri dari 4 perlakuan yaitu A  = Ikan disuntik dengan ekstrak daun sirih konsentrasi 75%, B  = Ikan disuntik dengan ekstrak daun sirih konsentrasi 25%, C  = Kontrol positif (ikan disuntik dengan akuades steril dan D  = Kontrol negatif (ikan tidak disuntik, diulang 3 kali. Hasil uji sensitivitas antibakteri daun sirih yang mempunyai daya hambat dan daya bunuh paling besar terhadap bakteri A. hydrophila adalah ekstrak daun sirih-metanol. Pengujian MIC menunjukkan bahwa ekstrak daun sirih-metanol memiliki daya hambat minimal 25 % terhadap aktivitas bakteri A. hydrophila. Hasil uji toksisitas yang dilakukan terhadap ikan patin dengan konsentrasi 75% dan 25% menunjukkan bahwa mortalitas yang terjadi tidak mencapai 50%. Pengamatan hematologis pada masing-masing perlakuan menunjukkan hasil yang berpengaruh tidak nyata terhadap kesehatan ikan patin. Parameter kualitas air pada penelitian ini yaitu, , kadar oksigen terlarut,  pH, amoniak, CO2 dan suhu masih dapat mendukung kehidupan ikan patin. This research was aimed a finding the part of Piper betle Linn  which had the biggest resistance to Aeromonas hydrophila bacteria and to know the minimal concentrate which could obstruct the growth of A. hydrophila  bacteria and to know effective concentrate toxicity of P. betle Linn  to Pangasius hypophthalmus.  The random sampling used proportionate stratified random sampling. In toxicity test, it had be done 4 treatment, which was given to fish, those were : A = fish was injected with 25 % concentrate of extract  P

Effects of external radiation in a co-culture model of endothelial cells and adipose-derived stem cells

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Haubner, Frank; Leyh, Michaela; Ohmann, Elisabeth; Pohl, Fabian; Prantl, Lukas; Gassner, Holger G

2013-01-01

The inflammatory response clinically observed after radiation has been described to correlate with elevated expression of cytokines and adhesion molecules by endothelial cells. Therapeutic compensation for this microvascular compromise could be an important approach in the treatment of irradiated wounds. Clinical reports describe the potential of adipose-derived stem cells to enhance wound healing, but the underlying cellular mechanisms remain largely unclear. Human dermal microvascular endothelial cells (HDMEC) and human adipose-derived stem cells (ASC) were cultured in a co-culture setting and irradiated with sequential doses of 2 to 12 Gy. Cell count was determined 48 h after radiation using a semi-automated cell counting system. Levels of interleukin-6 (IL-6), basic fibroblast growth factor (FGF), intercellular adhesion molecule-1 (ICAM-1) and vascular cell adhesion molecule-1 (VCAM-1) were determined in the supernatants using enzyme-linked immunosorbent assay (ELISA). Irradiated HDMEC and ASC as well as non-irradiated co-cultures, HDMEC or ASC respectively were used as controls. Cell count was significantly reduced in irradiated co-cultures of HDMEC and ASC compared to non-irradiated controls. Levels of IL-6, FGF, ICAM-1 and VCAM-1 in the supernatants of the co-cultures were significantly less affected by external radiation in comparison to HDMEC. The increased expression of cytokines and adhesion molecules by HDMEC after external radiation is mitigated in the co-culture setting with ASC. These in vitro changes seem to support the clinical observation that ASC may have a stabilizing effect when injected into irradiated wounds

The effects of biomimetically conjugated VEGF on osteogenesis and angiogenesis of MSCs (human and rat) and HUVECs co-culture models.

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Lü, Lanxin; Deegan, Anthony; Musa, Faiza; Xu, Tie; Yang, Ying

2018-07-01

The purpose of this work was to investigate if the biomimetically conjugated VEGF and HUVECs co-culture could modulate the osteogenic and angiogenic differentiation of MSCs derived from rat and human bone marrow (rMSCs and hMSCs). After treated by ammonia plasma, Poly(lactic-co-glycolic acid) (PLGA) electrospun nanofibers were immobilized with VEGF through heparin to fulfil the sustained release. The proliferation capacity of rMSCs and hMSCs on neat PLGA nanofibers (NF) and VEGF immobilized NF (NF-VEGF) surfaces were assessed by CCK-8 and compared when MSCs were mono-cultured and co-cultured with HUVECs. The effect of VEGF and HUVECs co-culturing on osteogenic and angiogenic differentiation of rMSCs and hMSCs were investigated by calcium deposits and CD31 expression on NF and NF-VEGF surfaces. The results indicated that VEGF has been biomimetically immobilized onto PLGA nanofibers surface and kept sustained release successfully. The CD31 staining results showed that both VEGF and HUVECs co-culture could enhance the angiogenesis of rMSCs and hMSCs. However, the proliferation and osteogenic differentiation of MSCs when cultured with VEGF and HUVECs showed a species dependent response. Taken together, VEGF immobilization and co-culture with HUVECs promoted angiogenesis of MSCs, indicating a good strategy for vascularization in bone tissue engineering. Copyright © 2018. Published by Elsevier B.V.

Concentração inibitória mínima de oxitetraciclina para isolados de Aeromonas Hydrophila obtidos de diferentes fontes Minimal inhibitory concentration to oxitetracycline in Aeromonas Hydrophila strains isolated from different sources

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Delton José Pereira Júnior

2006-12-01

Full Text Available Objetivou-se com este trabalho determinar a concentração inibitória mínima (MIC de oxitetraciclina para isolados de Aeromonas hydrophila obtidos de pescado, água de cultivo de peixes e casos de septicemia hemorrágica em peixes. Foi determinado MIC de 100 isolados de A. hydrophila, oriundos de 12 pisciculturas localizadas nos Estados de Minas Gerais, Rio de Janeiro e Rio Grande do Sul, utilizando a técnica de macrodiluição em caldo. Os resultados demonstraram que 14 isolados apresentaram MIC>100 µg/mL (resistentes e 86 apresentaram MICThe aim of this paper was to determine the minimal inhibitory concentration (MIC to oxitetracycline in Aeromonas hydrophila strains isolated from marketed fish, pond water of piscicultures and fish suffering hemorrhagic septicemia. These strains were obtained from 12 different piscicultures from Minas Gerais, Rio de Janeiro e Rio Grande do Sul states. It was determined the MIC to 100 strains, using the broth macrodilution method, and the results showed that 86 strains shown value of MIC100µg/mL (classified as resistant. No differences were observed among strains isolated from marketed fish, pond water and disease outbreaks. I was concluded that the variation detected may represent a risk of selection of resistant bacterial strains in aquatic environments under use of oxitetracycline.

Decolorisation of Congo Red by Aeromonas hydrophila Under Microaerophilic Conditions

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El Gizawy, S.; Refae, R.A.; Abd El Kareem, H.; Gomaa, O.M.; Hamed, H.

2009-01-01

Among 22 local bacterial isolates, Aeromonas hydrophila isolated from Bolti fish showed a remarkable ability to decolorize congo red under microaerophilic conditions (98.8% in 48 hrs) without prior adaptation. The bacterium had the ability to grow and decolorized high concentrations of congo red which varied from 50 to 1000 mg/l , the decolorisation was irrelevant to bacterial growth. The strain was capable of decolorizing congo red under a wide range of temperatures (20-40 degree C), ph (3-8), NaCl (0-200 mg/l) and also had the ability to decolorized mono, di and poly azo dyes. This bacterium is notable in its ability to decolorized the diazo congo red under extreme conditions of temperature, ph, salinity and high dye concentrations which makes it suitable for decolorisation under extreme industrial conditions

Expression, purification, crystallization and preliminary X-ray analysis of Aeromonas hydrophilia metallo-[beta]-lactamase

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Sharma, N.; Toney, J.H.; Fitzgerald, P.M.D. (Merck)

2010-07-20

The CphA metallo-{beta}-lactamase from Aeromonas hydrophilia has been expressed, purified and crystallized by the hanging-drop vapor-diffusion method using ammonium sulfate as the precipitant. The crystals exhibit orthorhombic symmetry (P2{sub 1}2{sub 1}2), with unit-cell parameters a = 40.75, b = 42.05, c = 128.88 {angstrom}. There is one monomer in the asymmetric unit and the solvent content is estimated to be 44% by volume. A data set extending to 1.8 {angstrom} has been measured.

A New In Vitro Co-Culture Model Using Magnetic Force-Based Nanotechnology.

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Takanari, Hiroki; Miwa, Keiko; Fu, XianMing; Nakai, Junichi; Ito, Akira; Ino, Kousuke; Honda, Hiroyuki; Tonomura, Wataru; Konishi, Satoshi; Opthof, Tobias; van der Heyden, Marcel Ag; Kodama, Itsuo; Lee, Jong-Kook

2016-10-01

Skeletal myoblast (SkMB) transplantation has been conducted as a therapeutic strategy for severe heart failure. However, arrhythmogenicity following transplantation remains unsolved. We developed an in vitro model of myoblast transplantation with "patterned" or "randomly-mixed" co-culture of SkMBs and cardiomyocytes enabling subsequent electrophysiological, and arrhythmogenic evaluation. SkMBs were magnetically labeled with magnetite nanoparticles and co-cultured with neonatal rat ventricular myocytes (NRVMs) on multi-electrode arrays. SkMBs were patterned by a magnet beneath the arrays. Excitation synchronicity was evaluated by Ca(2+) imaging using a gene-encoded Ca(2+) indicator, G-CaMP2. In the monoculture of NRVMs (control), conduction was well-organized. In the randomly-mixed co-culture of NRVMs and SkMBs (random group), there was inhomogeneous conduction from multiple origins. In the "patterned" co-culture where an en bloc SKMB-layer was inserted into the NRVM-layer, excitation homogenously propagated although conduction was distorted by the SkMB-area. The 4-mm distance conduction time (CT) in the random group was significantly longer (197 ± 126 ms) than in control (17 ± 3 ms). In the patterned group, CT through NRVM-area did not change (25 ± 3 ms), although CT through the SkMB-area was significantly longer (132 ± 77 ms). The intervals between spontaneous excitation varied beat-to-beat in the random group, while regular beating was recorded in the control and patterned groups. Synchronized Ca(2+) transients of NRVMs were observed in the patterned group, whereas those in the random group were asynchronous. Patterned alignment of SkMBs is feasible with magnetic nanoparticles. Using the novel in vitro model mimicking cell transplantation, it may become possible to predict arrhythmogenicity due to heterogenous cell transplantation. J. Cell. Physiol. 231: 2249-2256, 2016. © 2016 Wiley Periodicals, Inc. © 2016 Wiley Periodicals, Inc.

Genome Sequence of the Melanin-Producing Extremophile Aeromonas salmonicida subsp. pectinolytica Strain 34melT

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Pavan, María Elisa; Pavan, Esteban E.; López, Nancy Irene; Levin, Laura Noemí; Pettinari, María Julia

2017-01-01

The genome of Aeromonas salmonicida subsp. pectinolytica strain 34melT, isolated from a heavily polluted river, contains several genomic islands and putative virulence genes. The identification of genes involved in resistance to different kinds of stress sheds light on the mechanisms used by this strain to thrive in an extreme environment. Fil: Pavan, María Elisa. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales. Departamento de Química Biológica; Argentina Fil: Pav...

Effects of naturally occurring aquatic organic fractions on 241Am uptake by Scenedesmus obliquus (Chlorophyceae) and Aeromonas hydrophila (Pseudomonadaceae)

International Nuclear Information System (INIS)

Giesy, J.P. Jr.; Paine, D.

1977-01-01

Naturally occurring organics were extracted from water collected from Skinface Pond near Aiken, S.C. Organics were separated into four nominal diameter size fractions (I, greater than 0.0183; II, 0.0183 to 0.0032; III, 0.0032 to 0.0009; IV, less than 0.0009 μm) by membrane ultrafiltration and introduced into Scenedesmus obliquus and Aeromonas hydrophila cultures to determine their effects on 241 Am availability for uptake. Effects on 241 Am uptake were determined in actively growing S. obliquus cultures after 96 h of growth and in dense cultures of nongrowing cells after 4 h. Uptake by A. hydrophila was determined after 4 and 24 h in actively growing cultures. All organic fractions stimulated S. obliquus growth, with the most pronounced effects due to larger organic fractions, whereas no apparent growth stimulation of A. hydrophila was observed for any organic fraction. For both long-term and short-term studies, cellular 241 Am concentration (picocuries/cell) increased with increasing 241 Am concentration for S. obliquus and A. hydrophila. Fraction IV increased 241 Am uptake by both S. obliquus and A. hydrophila during 4-h incubations. During 96-h incubations fraction I was flocculated and cosedimented, with S. obliquus and A. hydrophila cells causing an apparent increase in 241 Am uptake. Fractions II and III reduced apparent 241 Am uptake by S. obliquus as a result of biological dilution caused by increased algal growth due to the organics. Fraction IV caused a reduction in 241 Am uptake by S. obliquus not attributable to biological dilution. Organics increased 241 Am uptake by A. hydrophila during 4- and 24-h incubations. A. hydrophila also caused flocculation of fraction I during 96-h incubations

Co-Cultures of Pseudomonas aeruginosa and Roseobacter denitrificans Reveal Shifts in Gene Expression Levels Compared to Solo Cultures

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Crystal A. Conway

2012-01-01

Full Text Available Consistent biosynthesis of desired secondary metabolites (SMs from pure microbial cultures is often unreliable. In a proof-of-principle study to induce SM gene expression and production, we describe mixed “co-culturing” conditions and monitoring of messages via quantitative real-time PCR (qPCR. Gene expression of model bacterial strains (Pseudomonas aeruginosa PAO1 and Roseobacter denitrificans Och114 was analyzed in pure solo and mixed cocultures to infer the effects of interspecies interactions on gene expression in vitro, Two P. aeruginosa genes (PhzH coding for portions of the phenazine antibiotic pathway leading to pyocyanin (PCN and the RhdA gene for thiosulfate: cyanide sulfurtransferase (Rhodanese and two R. denitrificans genes (BetaLact for metallo-beta-lactamase and the DMSP gene for dimethylpropiothetin dethiomethylase were assessed for differential expression. Results showed that R. denitrificans DMSP and BetaLact gene expression became elevated in a mixed culture. In contrast, P. aeruginosa co-cultures with R. denitrificans or a third species did not increase target gene expression above control levels. This paper provides insight for better control of target SM gene expression in vitro and bypass complex genetic engineering manipulations.

PEMBERIAN PROBIOTIK LACTOBACILLUS BREVIS DAN PREBIOTIK OLIGOSAKARIDA PADA BENIH PATIN SIAM (Pangasionodon hypophthalmus YANG DIINFEKSI Aeromonas hydrophila

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Yuke Eliyani

2013-08-01

Lactobacillus brevis, prebiotik oligosakarida dan sinbiotiknya terhadap jumlah bakteri Lactobacillus sp. dan total bakteri dalam usus, total eritrosit, total leukosit, diferensial leukosit, aktivitas fagositik, sintasan, tingkat pertumbuhan, serta FCR benih ikan patin siam yang diinfeksi Aeromonas hydrophila. Hasil uji karakteristik menunjukkan bahwa jenis probiotik dan patogen adalah Lactobacillus sp. dan Aeromonas hydrophila. Pada uji in vivo digunakan lima perlakuan yang terdiri atas K(+, K(-, probiotik (pro, prebiotik (pre serta sinbiotik (sin. Bakteri Lactobacillus sp. ditemukan di usus pada perlakuan probiotik dan sinbiotik dengan kisaran jumlah sekitar 101 sampai 106 (CFU/g. Total eritrosit, total leukosit, aktivitas fagositik berbeda nyata (P<0,05 dengan kontrol pada beberapa waktu pengamatan. Tingkat sintasan terendah diperoleh pada perlakuan K(+ sebesar 43,33±11,55; sedangkan empat perlakuan lainnya memperoleh nilai 100%. Tingkat pertumbuhan harian berbeda nyata antar perlakuan, nilai terbaik dicapai pada perlakuan pemberian sinbiotik sebesar 3,370±0,14. Nilai FCR perlakuan probiotik, prebiotik dan sinbiotik menunjukkan beda nyata dengan kontrol. Perlakuan sin, pre, pro memberikan nilai yang lebih baik pada total eritrosit, total leukosit, aktivitas fagositik, sintasan, pertumbuhan, dan FCR dibandingkan kontrol.

Development of highly sensitive electrochemical genosensor based on multiwalled carbon nanotubes-chitosan-bismuth and lead sulfide nanoparticles for the detection of pathogenic Aeromonas.

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Fernandes, António Maximiano; Abdalhai, Mandour H; Ji, Jian; Xi, Bing-Wen; Xie, Jun; Sun, Jiadi; Noeline, Rasoamandrary; Lee, Byong H; Sun, Xiulan

2015-01-15

In this paper, we reported the construction of new high sensitive electrochemical genosensor based on multiwalled carbon nanotubes-chitosan-bismuth complex (MWCNT-Chi-Bi) and lead sulfide nanoparticles for the detection of pathogenic Aeromonas. Lead sulfide nanoparticles capped with 5'-(NH2) oligonucleotides thought amide bond was used as signalizing probe DNA (sz-DNA) and thiol-modified oligonucleotides sequence was used as fixing probe DNA (fDNA). The two probes hybridize with target Aeromonas DNA (tDNA) sequence (fDNA-tDNA-szDNA). The signal of hybridization is detected by differential pulse voltammetry (DPV) after electrodeposition of released lead nanoparticles (PbS) from sz-DNA on the surface of glass carbon electrode decorated with MWCNT-Chi-Bi, which improves the deposition and traducing electrical signal. The optimization of incubation time, hybridization temperature, deposition potential, deposition time and the specificity of the probes were investigated. Our results showed the highest sensibility to detect the target gene when compared with related biosensors and polymerase chain reaction (PCR). The detection limit for this biosensor was 1.0×10(-14) M. We could detect lower than 10(2) CFU mL(-1) of Aeromonas in spiked tap water. This method is rapid and sensitive for the detection of pathogenic bacteria and would become a potential application in biomedical diagnosis, food safety and environmental monitoring. Copyright © 2014 Elsevier B.V. All rights reserved.

Development of striatal patch/matrix organization in organotypic co-cultures of perinatal striatum, cortex and substantia nigra.

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Snyder-Keller, A; Costantini, L C; Graber, D J

2001-01-01

Organotypic cultures of fetal or early postnatal striatum were used to assess striatal patch formation and maintenance in the presence or absence of dopaminergic and glutamatergic influences. Vibratome-cut slices of the striatum prepared from embryonic day 19 to postnatal day 4 rat pups were maintained in static culture on clear membrane inserts in Dulbecco's modified Eagle's medium/F12 (1:1) with 20% horse serum. Some were co-cultured with embryonic day 12-16 ventral mesencephalon and/or embryonic day 19 to postnatal day 4 cortex, which produced a dense dopaminergic innervation and a modest cortical innervation. Donors of striatal and cortical tissue were previously injected with bromo-deoxyuridine (BrdU) on embryonic days 13 and 14 in order to label striatal neurons destined to populate the patch compartment of the striatum. Patches of BrdU-immunoreactive cells were maintained in organotypic cultures of late prenatal (embryonic days 20-22) or early postnatal striatum in the absence of nigral dopaminergic or cortical glutamatergic influences. In slices taken from embryonic day 19 fetuses prior to the time of in vivo patch formation, patches were observed to form after 10 days in vitro, in 39% of nigral-striatal co-cultures compared to 6% of striatal slices cultured alone or in the presence of cortex only. Patches of dopaminergic fibers, revealed by tyrosine hydroxylase immunoreactivity, were observed in the majority of nigral-striatal co-cultures. Immunostaining for the AMPA-type glutamate receptor GluR1 revealed a dense patch distribution in nearly all cultures, which developed in embryonic day 19 cultures after at least six days in vitro. These findings indicate that striatal patch/matrix organization is maintained in organotypic culture, and can be induced to form in vitro in striatal slices removed from fetuses prior to the time of in vivo patch formation. Furthermore, dopaminergic innervation from co-cultured pieces of ventral mesencephalon enhances patch

Transmissão do Trypanosoma cruzi em três gerações de Cavia porcellus sem a participação de triatomíneos

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Ítalo A. Sherlock

1976-02-01

Full Text Available Foi verificada a transmissão sucessiva do T. cruzi em três gerações da cobaia Cavia porcellus sem a participação de triatomíneos. Embora não fosse determinado qual das vias, se placentária, leite, excreções ou contágio direto pelo qual o protozoário foi transmitido para os descendentes, chama-se atenção para a importância da manutenção de reservatórios da Doença de Chagas, mesmo na ausência de vetores invertebrados.

Effect of co-culture with enterocinogenic E. faecium on L. monocytogenes key virulence gene expression

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Eleftherios H. Drosinos

2016-08-01

Full Text Available The aim of the present study was to assess the expression of key virulence genes during co-culture of L. monocytogenes with a bacteriocinogenic E. faecium strain in liquid growth medium. For that purpose, BHI broth was inoculated with 7 log CFU·mL–1 L. monocytogenes and 4, 5 or 6 log CFU·mL–1 E. faecium. Sampling took place after 8 and 24 h of incubation, corresponding to the maximum and minimum of enterocin production, respectively. The RNA was extracted, stabilized and expression of prfA, sigB, hly, plcA, plcB, inlA, inlB, inlC and inlJ, was assessed by RT-qPCR. Most of the genes were downregulated during co-culture at 5 °C. Moreover, a statistically significant effect of the inoculum level was evident in most of the cases. On the contrary, no effect on the transcription level of most of the genes was observed during co-culture at 37 °C.

Vaccination in Nile tilapia broodstock with whole cell vaccine and disease resistance in its fry against Aeromonas hydrophila

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Sukenda Sukenda

2017-07-01

Full Text Available ABSTRACT The aim of this study was to analyze the effectivity of vaccination in Nile tilapia broodstock with whole cell vaccine and disease resistance in fry tilapia against Aeromonas hydrophila. Tilapia Nirwana strain that used for this had average body weight of 185±13.23 g and were maintained in ponds sizing of (2.5×2.5×1 m3. Vaccinations that has been done through intraperitoneal injection using dose of 0.1 mL/fish, meanwhile the fish for control was injected by phosphate buffered saline (PBS. This study used complete randomized design with two treatments and three replications. Antibody level was measured by using indirect enzyme-linked immunosorbent assay (ELISA method in the broodstock, egg, and fry.  Challenge test in fry tilapia performed at the age of 5, 10, and 15 days. The results showed that vaccination in tilapia broodstock delivered a significant antibody level in broodstock, eggs, and fry (P<0.05 compared to the control. Relative percent survival of offspring at 5, 10, and 15 days were 78.26%, 70.59%, and 65.52%, respectively.  As a conclusion, vaccination in tilapia broodstock was effective to improve specific and non-specific immunity, and protect fry tilapia from A. hydrophila infection through maternal immunity. Keywords: vaccination, antibody, maternal immunity, tilapia, Aeromonas hydrophila  ABSTRAK Penelitian ini bertujuan untuk menganalisis efikasi vaksinasi pada induk nila dengan vaksin sel utuh dan ketahanan benih yang dihasilkan terhadap Aeromonas hydrophila. Ikan nila stain Nirwana yang digunakan dalam penelitian memiliki bobot rata-rata 185±13,23 g dan ikan dipelihara dalam kolam (2,5×2,5×1 m3. vaksinasi dilakukan melalui penyuntikan intraperitoneal dengan dosis 0,1 mL/ikan, sementara itu ikan kontrol disuntik dengan phosphate buffered saline (PBS. Penelitian ini menggunakan rancangan acak lengkap dengan dua perlakuan dan tiga ulangan. Tingkat antibodi diukur dengan menggunakan metode indirect enzyme

PEMBERIAN TOTAL FENOL TERIPANG PASIR (Holothuria scabra UNTUK MENINGKATKAN LEUKOSIT DAN DIFERENSIAL LEUKOSIT IKAN MAS (Cyprinus carpio YANG DIINFEKSI BAKTERI Aeromonas Hydrophila

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Achmad Suhermanto

2011-10-01

Full Text Available Freshwater fishery commodities are most likely to be increased production of carp (Cyprinus carpio. Intensification of cultivation which raises new problems with fish disease outbreaks caused by Aeromonas hydrophila. Providing of immunostimulatory bioactive compounds of sea cucumbers (Holothuria scabra can enhance non-specific immune responses in the goldfish as a mechanism of defense against disease. The purpose of this study is to determine the role and optimal dose of total phenols sea cucumber in enhancing non-specific immune response seen from the hematology parameters Extraction of bioactive components from sea cucumber prepared by using methanol and fractionated with ethyl acetate solvent (v/v. Identification of total phenols in ethyl acetate fraction performed using UV-Vis spectrophotometer and infrared. Total phenol produced was tested on carp with intraperitonial injection, the concentration of 0.09, 0.18 and 0.27 mg phenol/kg fish. Challenge test was done by using the Aeromonas hydrophila (107 cells/ml through the immersion method. The observations included hematological parameters and total plasma proteins.The observations of hematological parameters that Leukocytes, Neutrophils pre-and post-infection were significantly increased. Lymphocyte pre infection was increased significantly and post-bacterial infection was decreased significantly. Eosinophils, Monocytes pre-and post-bacterial infection were not differ significantly between treatment. The use of phenolic compounds at 0.09 mg / kg can be increase of non-specific immune response in carp (Cyprinus carpio. Key Words : Total phenol, Aeromonas hydrophila, Cyprinus carpio, nonspecific immune response 

Prevalence of listeria, Aeromonas, and Vibrio species in fish used for human consumption in Turkey.

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Yücel, Nihal; Balci, Senay

2010-02-01

A total of 78 raw retail fish samples from 30 freshwater and 48 marine fish were examined for the presence of Listeria, Aeromonas, and Vibrio species. The overall incidence of Listeria spp. was 30% in freshwater samples and 10.4% in marine fish samples. Listeria monocytogenes (44.5%) was the most commonly isolated species in freshwater fish, and Listeria murrayi (83.5%) was the most commonly isolated species in marine fish samples. Motile aeromonads were more common in marine fish samples (93.7%) than in freshwater fish samples (10%). Vibrio alginolyticus, Vibrio fluvialis, and Vibrio damsela were isolated only in marine fish samples, representing 40.9, 38.6, and 36.3% of Vibrio isolates, respectively. In freshwater and marine fish, the highest incidences of Listeria and Aeromonas were found in skin samples; the highest incidence of Vibrio in marine fish was found in gill samples. The location of Listeria spp. and L. monocytogenes in a fish was significantly different among freshwater fish. A high incidence of these bacterial pathogens was found in the brown trout (Salmo trutta) and horse mackerel (Trachurus trachurus). Handling of contaminated fish, cross-contamination, or eating raw fish might pose a health hazard, especially in immunosuppressed individuals, elderly people, and children. This study highlights the importance of bacterial pathogens in fish intended for human consumption, but more study is needed.

Cryopreservation studies of an artificial co-culture between the cobalamin-requiring green alga Lobomonas rostrata and the bacterium Mesorhizobium loti.

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Ridley, Christian J A; Day, John G; Smith, Alison G

2018-01-01

Algal-bacterial co-cultures, rather than cultures of algae alone, are regarded as having the potential to enhance productivity and stability in industrial algal cultivation. As with other inocula in biotechnology, to avoid loss of production strains, it is important to develop preservation methods for the long-term storage of these cultures, and one of the most commonly used approaches is cryopreservation. However, whilst there are many reports of cryopreserved xenic algal cultures, little work has been reported on the intentional preservation of both algae and beneficial bacteria in xenic cultures. Instead, studies have focused on the development of methods to conserve the algal strain(s) present, or to avoid overgrowth of bacteria in xenic isolates during the post-thaw recovery phase. Here, we have established a co-cryopreservation method for the long-term storage of both partners in a unialgal-bacterial co-culture. This is an artificial model mutualism between the alga Lobomonas rostrata and the bacterium Mesorhizobium loti , which provides vitamin B 12 (cobalamin) to the alga in return for photosynthate. Using a Planer Kryo 360 controlled-rate cooler, post-thaw viability (PTV) values of 72% were obtained for the co-culture, compared to 91% for the axenic alga. The cultures were successfully revived after 6 months storage in liquid nitrogen, and continued to exhibit mutualism. Furthermore, the alga could be cryopreserved with non-symbiotic bacteria, without bacterial overgrowth occurring. It was also possible to use less controllable passive freezer chambers to cryopreserve the co-cultures, although the PTV was lower. Finally, we demonstrated that an optimised cryopreservation method may be used to prevent the overgrowth potential of non-symbiotic, adventitious bacteria in both axenic and co-cultures of L. rostrata after thawing.

Assessment of Skin Pathological Responses in the Yellowfin Seabream (Acanthopagrus latus under the Aeromonas hydrophila Exposure

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Fatemeh Azadbakht

2018-01-01

Full Text Available Background: Bacterial diseases in cultured fish are considered the main problem to aquaculture system. Skin is the structure that covers the body in fish. Skin histopatological alterations were used to assess the effects of Aeromonas hydrophila exposure on the yellowfin seabream Acanthopagrus latus(. Methods: In this regard, 90 A. latus were exposed to sublethal concentrations of A. hydrophila (103,106 CFU/ml for 3 weeks. Results: Some more severe alternations found in the skin of fish exposed. The most frequent histopathological changes detected in the skin including hyperplasia of epidermis, hypertrophy and hyperplasia of the mucosal cells and dermis edema. Some more severe alternations found in the skin of fish exposed to higher level of A. hydrophila (106 CFU/ml included telangiectasia of dermis layer. In addition, according to the results of histometrical studies in treated fish compared to control group showed that thickness of epidermis and dermis layers were increased significantly (P<0.05. Conclusion: A. hydrophila can cause major histophatological changes in the skin of A. latus. In addition, histopathological changes of the skin provide helpful information about the environmental conditions and as particular biomarkers may provide imminent into evaluating the general health and stress status of fish.

Skin equivalent tissue-engineered construct: co-cultured fibroblasts/ keratinocytes on 3D matrices of sericin hope cocoons.

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Sunita Nayak

Full Text Available The development of effective and alternative tissue-engineered skin replacements to autografts, allografts and xenografts has became a clinical requirement due to the problems related to source of donor tissue and the perceived risk of disease transmission. In the present study 3D tissue engineered construct of sericin is developed using co-culture of keratinocytes on the upper surface of the fabricated matrices and with fibroblasts on lower surface. Sericin is obtained from "Sericin Hope" silkworm of Bombyx mori mutant and is extracted from cocoons by autoclave. Porous sericin matrices are prepared by freeze dried method using genipin as crosslinker. The matrices are characterized biochemically and biophysically. The cell proliferation and viability of co-cultured fibroblasts and keratinocytes on matrices for at least 28 days are observed by live/dead assay, Alamar blue assay, and by dual fluorescent staining. The growth of the fibroblasts and keratinocytes in co-culture is correlated with the expression level of TGF-β, b-FGF and IL-8 in the cultured supernatants by enzyme-linked immunosorbent assay. The histological analysis further demonstrates a multi-layered stratified epidermal layer of uninhibited keratinocytes in co-cultured constructs. Presence of involucrin, collagen IV and the fibroblast surface protein in immuno-histochemical stained sections of co-cultured matrices indicates the significance of paracrine signaling between keratinocytes and fibroblasts in the expression of extracellular matrix protein for dermal repair. No significant amount of pro inflammatory cytokines (TNF-α, IL-1β and nitric oxide production are evidenced when macrophages grown on the sericin matrices. The results all together depict the potentiality of sericin 3D matrices as skin equivalent tissue engineered construct in wound repair.

Microalgae Harvest through Fungal Pelletization—Co-Culture of Chlorella vulgaris and Aspergillus niger

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Sarman Oktovianus Gultom

2014-07-01

Full Text Available Microalgae harvesting is a labor- and energy-intensive process and new approaches to harvesting microalgae need to be developed in order to decrease the costs. In this study; co-cultivatation of filamentous fungus (Aspergillus niger and microalgae (Chlorella vulgaris to form cell pellets was evaluated under different conditions, including organic carbon source (glucose; glycerol; and sodium acetate concentration; initial concentration of fungal spores and microalgal cells and light. Results showed that 2 g/L of glucose with a 1:300 ratio of fungi to microalgae provided the best culturing conditions for the process to reach >90% of cell harvest efficiency. The results also showed that an organic carbon source was required to sustain the growth of fungi and form the cell pellets. The microalgae/fungi co-cultures at mixotrophic conditions obtained much higher total biomass than pure cultures of each individual strains; indicating the symbiotic relationship between two strains. This can benefit the microbial biofuel production in terms of cell harvest and biomass production.

Monocarboxylate transporter-dependent mechanism confers resistance to oxygen- and glucose-deprivation injury in astrocyte-neuron co-cultures.

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Gao, Chen; Zhou, Liya; Zhu, Wenxia; Wang, Hongyun; Wang, Ruijuan; He, Yunfei; Li, Zhiyun

2015-05-06

Hypoxic and low-glucose stressors contribute to neuronal death in many brain diseases. Astrocytes are anatomically well-positioned to shield neurons from hypoxic injury. During hypoxia/ischemia, lactate released from astrocytes is taken up by neurons and stored for energy. This process is mediated by monocarboxylate transporters (MCTs) in the central nervous system. In the present study, we investigated the ability of astrocytes to protect neurons from oxygen- and glucose-deprivation (OGD) injury via an MCT-dependent mechanism in vitro. Primary cultures of neurons, astrocytes, and astrocytes-neurons derived from rat hippocampus were subjected to OGD, MCT inhibition with small interfering (si)RNA. Cell survival and expression of MCT4, MCT2, glial fibrillary acidic protein, and neuronal nuclear antigen were evaluated. OGD significantly increased cell death in neuronal cultures and up-regulated MCT4 expression in astrocyte cultures, but no increased cell death was observed in neuron-astrocyte co-cultures or astrocyte cultures. However, neuronal cell death in co-cultures was increased by exposure to MCT4- or MCT2-specific siRNA, and this effect was attenuated by the addition of lactate into the extracellular medium of neuronal cultures prior to OGD. These findings demonstrate that resistance to OGD injury in astrocyte-neuron co-cultures occurs via an MCT-dependent mechanism. Copyright © 2015 Elsevier Ireland Ltd. All rights reserved.

Inhibition of MAPK and PKC pathways by 60Co γ-radiation in cultured vascular smooth muscle cells

International Nuclear Information System (INIS)

Jia Guanghong; Ma Yexin; Xiao Jianming

2002-01-01

Objective: To investigate the signal transduction pathways inhibited by 60 Co γ-radiation in cultured vascular smooth muscle cells (VSMC). Methods: The cultured VSMC were irradiated with 60 Co γ-radiation of 3.5, 7.0 and 14 Gy respectively. VSMC proliferation was measured by 3 H-TdR incorporation, while PKC, MAPK activities were determined by radioactivity assay. Results: Proliferation of VSMC was inhibited by 7.0, 14 Gy 60 Co γ-irradiation and the activities of PKC, MAPK were decreased significantly. Conclusion: Inhibitory effect of 7.0, 14 Gy 60 Co γ-irradiation on proliferation of VSMC might be resulted from decrease of the activity of PKC, MAPK

Temporally coordinated spiking activity of human induced pluripotent stem cell-derived neurons co-cultured with astrocytes.

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Kayama, Tasuku; Suzuki, Ikuro; Odawara, Aoi; Sasaki, Takuya; Ikegaya, Yuji

2018-01-01

In culture conditions, human induced-pluripotent stem cells (hiPSC)-derived neurons form synaptic connections with other cells and establish neuronal networks, which are expected to be an in vitro model system for drug discovery screening and toxicity testing. While early studies demonstrated effects of co-culture of hiPSC-derived neurons with astroglial cells on survival and maturation of hiPSC-derived neurons, the population spiking patterns of such hiPSC-derived neurons have not been fully characterized. In this study, we analyzed temporal spiking patterns of hiPSC-derived neurons recorded by a multi-electrode array system. We discovered that specific sets of hiPSC-derived neurons co-cultured with astrocytes showed more frequent and highly coherent non-random synchronized spike trains and more dynamic changes in overall spike patterns over time. These temporally coordinated spiking patterns are physiological signs of organized circuits of hiPSC-derived neurons and suggest benefits of co-culture of hiPSC-derived neurons with astrocytes. Copyright © 2017 Elsevier Inc. All rights reserved.

A Highly Efficient Human Pluripotent Stem Cell Microglia Model Displays a Neuronal-Co-culture-Specific Expression Profile and Inflammatory Response

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Walther Haenseler

2017-06-01

Full Text Available Microglia are increasingly implicated in brain pathology, particularly neurodegenerative disease, with many genes implicated in Alzheimer's, Parkinson's, and motor neuron disease expressed in microglia. There is, therefore, a need for authentic, efficient in vitro models to study human microglial pathological mechanisms. Microglia originate from the yolk sac as MYB-independent macrophages, migrating into the developing brain to complete differentiation. Here, we recapitulate microglial ontogeny by highly efficient differentiation of embryonic MYB-independent iPSC-derived macrophages then co-culture them with iPSC-derived cortical neurons. Co-cultures retain neuronal maturity and functionality for many weeks. Co-culture microglia express key microglia-specific markers and neurodegenerative disease-relevant genes, develop highly dynamic ramifications, and are phagocytic. Upon activation they become more ameboid, releasing multiple microglia-relevant cytokines. Importantly, co-culture microglia downregulate pathogen-response pathways, upregulate homeostatic function pathways, and promote a more anti-inflammatory and pro-remodeling cytokine response than corresponding monocultures, demonstrating that co-cultures are preferable for modeling authentic microglial physiology.

Trophic Effects of Mesenchymal Stem Cells in Chondrocyte Co-Cultures are Independent of Culture Conditions and Cell Sources

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Wu, Ling; Prins, H.J.; Helder, M.; van Blitterswijk, Clemens; Karperien, Hermanus Bernardus Johannes

2012-01-01

Earlier, we have shown that the increased cartilage production in pellet co-cultures of chondrocytes and bone marrow-derived mesenchymal stem cells (BM-MSCs) is due to a trophic role of the MSC in stimulating chondrocyte proliferation and matrix production rather than MSCs actively undergoing

Trophic effects of mesenchymal stem cells in chondrocyte co-cultures are independent of culture conditions and cell sources

NARCIS (Netherlands)

Wu, L.; Prins, H.J.; Helder, M.N.; van Blitterswijk, C.A.; Karperien, M.

2012-01-01

Earlier, we have shown that the increased cartilage production in pellet co-cultures of chondrocytes and bone marrow-derived mesenchymal stem cells (BM-MSCs) is due to a trophic role of the MSC in stimulating chondrocyte proliferation and matrix production rather than MSCs actively undergoing

Pancarditis in a patient with tropical pyomyositis due to Aeromonas infection

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Nikhil Gupta

2015-01-01

Full Text Available Tropical pyomyositis is a suppurative disease of the skeletal muscles. The most common site of the infection being muscles of the thigh. Staphylococcus aureus is the most common organism isolated. Aeromonas is one of the rarest organisms described as the etiologic agent for this disease. Rarely, cardiac involvement in pyomyositis has been described in the form of myocarditis and pyopericardium. Pancarditis is a very rare event in the natural course of pyomyositis. To the best of our knowledge, there is a single case report in world literature regarding involvement of all the layers of the heart in pyomyositis. We report a case of a 30-year-old immunocompetent male who had pyomyositis and developed pancarditis, which was managed successfully with conservative management.

Prevascularization of 3D printed bone scaffolds by bioactive hydrogels and cell co-culture.

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Kuss, Mitchell A; Wu, Shaohua; Wang, Ying; Untrauer, Jason B; Li, Wenlong; Lim, Jung Yul; Duan, Bin

2017-09-13

Vascularization is a fundamental prerequisite for large bone construct development and remains one of the main challenges of bone tissue engineering. Our current study presents the combination of 3D printing technique with a hydrogel-based prevascularization strategy to generate prevascularized bone constructs. Human adipose derived mesenchymal stem cells (ADMSC) and human umbilical vein endothelial cells (HUVEC) were encapsulated within our bioactive hydrogels, and the effects of culture conditions on in vitro vascularization were determined. We further generated composite constructs by forming 3D printed polycaprolactone/hydroxyapatite scaffolds coated with cell-laden hydrogels and determined how the co-culture affected vascularization and osteogenesis. It was demonstrated that 3D co-cultured ADMSC-HUVEC generated capillary-like networks within the porous 3D printed scaffold. The co-culture systems promoted in vitro vascularization, but had no significant effects on osteogenesis. The prevascularized constructs were subcutaneously implanted into nude mice to evaluate the in vivo vascularization capacity and the functionality of engineered vessels. The hydrogel systems facilitated microvessel and lumen formation and promoted anastomosis of vascular networks of human origin with host murine vasculature. These findings demonstrate the potential of prevascularized 3D printed scaffolds with anatomical shape for the healing of larger bone defects. © 2017 Wiley Periodicals, Inc. J Biomed Mater Res Part B: Appl Biomater, 2017. © 2017 Wiley Periodicals, Inc.

Lactobacillus plantarum (VR1 isolated from an Ayurvedic medicine (Kutajarista ameliorates in vitro cellular damage caused by Aeromonas veronii

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Patole Milind S

2011-06-01

Full Text Available Abstract Background Lactobacillus plantarum is considered as a safe and effective probiotic microorganism. Among various sources of isolation, traditionally fermented foods are considered to be rich in Lactobacillus spp., which can be exploited for their probiotic attribute. Antibacterial property of L. plantarum has been demonstrated against various enteric pathogens in both in vitro and in vivo systems. This study was aimed at characterizing L. plantarum isolated from Kutajarista, an ayurvedic fermented biomedicine, and assessing its antagonistic property against a common enteropathogen Aeromonas veronii. Results We report the isolation of L. plantarum (VR1 from Kutajarista, and efficacy of its cell free supernatant (CFS in amelioration of cytotoxicity caused by Aeromonas veronii. On the part of probiotic attributes, VR1 was tolerant to pH 2, 0.3% bile salts and simulated gastric juice. Additionally, VR1 also exhibited adhesive property to human intestinal HT-29 cell line. Furthermore, CFS of VR1 was antibacterial to enteric pathogens like Pseudomonas aeruginosa, Staphylococcus aureus, Escherichia coli, Aeromonas veronii and clinical isolates of P. aeruginosa and E. coli. Detailed study regarding the effect of VR1 CFS on A. veronii cytotoxicity showed a significant decrease in vacuole formation and detrimental cellular changes in Vero cells. On the other hand, A. veronii CFS caused disruption of tight junction proteins ZO-1 and actin in MDCK cell line, which was prevented by pre-incubation with CFS of VR1. Conclusions This is the first study to report isolation of L. plantarum (VR1 from Kutajarista and characterisation for its probiotic attributes. Our study demonstrates the antagonistic property of VR1 to A. veronii and effect of VR1 CFS in reduction of cellular damage caused by A. veronii in both Vero and MDCK cell lines.

Hematologia e sinais clínicos de Piaractus mesopotamicus infectados experimentalmente com Aeromonas hydrophila = Hematology and clinical signs of Piaractus mesopotamicus experimentally infected with Aeromonas hydrophila

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Flávio Ruas Moraes

2009-01-01

Full Text Available Este ensaio teve como objetivo avaliar as respostas hematológicas e a ocorrência de sinais clínico-patológicos de Piaractus mesopotamicus infectados experimentalmente por Aeromonas hydrophila. Foram utilizados 324 pacus, P. mesopotamicus, com peso de 102,84 ±27,1 g, distribuídos em 27 caixas de 300 L de capacidade, sendo 14 peixes por caixa. Os peixes foram mantidos nas caixas durante um mês, recebendo dieta balanceada, período em que se evitou qualquer tipo de manejo estressante. Passado esse período, foram infectadoscom 6 x 106 UFC de A. hydrophila, injetada via intraperitoneal. As coletas e análises de sangue foram realizadas 24h antes da infecção experimental e 24h após a infecção. Os resultados permitem concluir que a infecção por A. hydrophila em P. mesopotamicus écaracterizada por escurecimento da derme e sinais clínicos-patológicos relacionados à hemorragia, anemia normocítica-hipocrômica, redução nos níveis de proteínas, globulinasplasmáticas e eosinófilos, leucopenia, linfopenia, trombocitopenia, neutrofilia e monocitose. The purpose of this study was to evaluate the hematological response and clinical sign of Piaractus mesopotamicus infected byAeromonas hydrophila. Were used 324 pacus, P. mesopotamicus weighing 102.84 ± 27.1 g distributed in 27 water tanks with 300 L of capacity, 14 fish per tank. The fish were kept in the water tanks during one month, receiving appropriate diet. In this period was avoidedstressing management. After this period, the fish were infected with 6 × 108 cfu of A. hydrophila fish-1, injected intraperitoneally. The blood collection was carried 24h before experimental infection and 24h after its. The A. hydrophila infection in P. mesopotamicus ischaracterized by dark skin and clinical signs related to hemorrhage, hypochromic normocytic anemia, decrease on plasmatic proteins and globulins levels, leucopenia, lymphopenia, thrombocytopenia, eosinophilia, neutrophilia and monocytosis.

Dopaminergic differentiation of human neural stem cells mediated by co-cultured rat striatal brain slices

DEFF Research Database (Denmark)

Anwar, Mohammad Raffaqat; Andreasen, Christian Maaløv; Lippert, Solvej Kølvraa

2008-01-01

differentiation, we co-cultured cells from a human neural forebrain-derived stem cell line (hNS1) with rat striatal brain slices. In brief, coronal slices of neonatal rat striatum were cultured on semiporous membrane inserts placed in six-well trays overlying monolayers of hNS1 cells. After 12 days of co......Properly committed neural stem cells constitute a promising source of cells for transplantation in Parkinson's disease, but a protocol for controlled dopaminergic differentiation is not yet available. To establish a setting for identification of secreted neural compounds promoting dopaminergic...

76 FR 58032 - Notice of Intent To Repatriate a Cultural Item: Denver Museum of Nature and Science, Denver, CO

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2011-09-19

... Denver Museum of Nature & Science, Denver, CO, that meets the definition of an object of cultural... Cultural Item: Denver Museum of Nature and Science, Denver, CO AGENCY: National Park Service, Interior. ACTION: Notice. SUMMARY: The Denver Museum of Nature & Science, in consultation with the appropriate...

Cryptosporidium homai n. sp. (Apicomplexa: Cryptosporidiiae) from the guinea pig (Cavia porcellus).

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Zahedi, Alireza; Durmic, Zoey; Gofton, Alexander W; Kueh, Susan; Austen, Jill; Lawson, Malcolm; Callahan, Lauren; Jardine, John; Ryan, Una

2017-10-15

The morphological, biological, and molecular characterisation of a new Cryptosporidium species from the guinea pig (Cavia porcellus) are described, and the species name Cryptosporidium homai n. sp. is proposed. Histological analysis conducted on a post-mortem sample from a guinea pig euthanised due to respiratory distress, identified developmental stages of C. homai n. sp. (trophozoites and meronts) along the intestinal epithelium. Molecular analysis at 18S rRNA (18S), actin and hsp70 loci was then conducted on faeces from an additional 7 guinea pigs positive for C. homai n. sp. At the 18S, actin and hsp70 loci, C. homai n. sp. exhibited genetic distances ranging from 3.1% to 14.3%, 14.4% to 24.5%, and 6.6% to 20.9% from other Cryptosporidium spp., respectively. At the 18S locus, C. homai n. sp. shared 99.1% similarity with a previously described Cryptosporidium genotype in guinea pigs from Brazil and it is likely that they are the same species, however this cannot be confirmed as actin and hsp70 sequences from the Brazilian guinea pig genotype are not available. Phylogenetic analysis of concatenated 18S, actin and hsp70 sequences showed that C. homai n. sp. exhibited 9.1% to 17.3% genetic distance from all other Cryptosporidium spp. This clearly supports the validity of C. homai n. sp. as a separate species. Copyright © 2017 Elsevier B.V. All rights reserved.

Effects of Electrical Stimulation in Sympathetic Neuron-Cardiomyocyte Co-cultures

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Takeuchi, Akimasa; Tani, Hiromasa; Mori, Masahide; Moriguchi, Hiroyuki; Kotani, Kiyoshi; Lee, Jong-Kook; Noshiro, Makoto; Jimbo, Yasuhiko

The sympathetic nervous system is one of the principal sources for regulating cardiovascular functions. Little is known, however, about the network-level interactions between sympathetic neurons and cardiomyocytes. In this study, a semi-separated co-culture system of superior cervical ganglion (SCG) neurons and ventricular myocytes (VMs) was developed by using a polydimethylsyloxane (PDMS) chamber placed on a microelectrode-array (MEA) substrate. Neurites of SCG neurons passed through a conduit of the chamber and reached VMs. Evoked activities of SCG neurons were observed from several electrodes immediately after applying constant-voltage stimulation (1 V, 1 ms, biphasic square pulses) to SCG neurons by using 32 electrodes. Furthermore, this stimulation was applied to SCG neurons at the frequency of 1, 5 and 10 Hz. After applying these three kinds of stimulations, mean minute contraction rate of VMs increased with an increase in the frequency of stimulation. These results suggest that changes in contraction rate of VMs after applying electrical stimulations to SCG neurons depend on frequencies of these stimulations and that the heart-regulating mechanisms as well as that in the body were formed in this co-culture system.

A New In Vitro Co-Culture Model Using Magnetic Force-Based Nanotechnology

NARCIS (Netherlands)

Takanari, Hiroki; Miwa, Keiko; Fu, Xianming; Nakai, Junichi; Ito, Akira; Ino, Kousuke; Honda, Hiroyuki; Tonomura, Wataru; Konishi, Satoshi; Opthof, Tobias; van der Heyden, Marcel Ag; Kodama, Itsuo; Lee, Jong-Kook

2016-01-01

Skeletal myoblast (SkMB) transplantation has been conducted as a therapeutic strategy for severe heart failure. However, arrhythmogenicity following transplantation remains unsolved. We developed an in vitro model of myoblast transplantation with "patterned" or "randomly-mixed" co-culture of SkMBs

Aeromonas hydrophila-associated skin lesions and septicaemia in a Nile crocodile (Crocodylus niloticus : clinical communication

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H. Turutoglu

2005-06-01

Full Text Available Aeromonas hydrophila is one of the most common bacteria associated with the aquatic environment. There are , however, limited data on A. hydrophila infection in crocodilians. The aim of this report is to describe a case of skin lesions and septicaemia associated with A. hydrophila in a Nile crocodile (Crocodylus niloticus. A captive male crocodile in the Zoological Park of Antalya (Turkey was found dead without showing signs of any disease. Gross examination showed brown or red-spotted skin lesions of varying size. These lesions were mostly scattered over the abdomen and occasionally on the tail and feet. At necropsy, numerous white, multifocal and randomly distributed areas were seen on the liver. Gram-stained smears from skin and liver lesions showed Gram-negative bacilli arranged in clusters. Pure cultures of A. hydrophila were recovered from skin, internal organs and blood. Isolates were found to be susceptible to ceftiofur, amoxicillin + clavulanic acid, oxytetracycline, enrofloxacin, danofloxacin, neomycin, gentamicin, and lincomycin + neomycin. A pathogenicity test was performed using this isolate on 4 male 2-year-old New Zealand white rabbits. Local abscesses formed in 2 rabbits injected subcutaneously and the 2 that were injected intraperitoneally died as a result of septicaemia. In conclusion, this report has shown that A. hydrophila may cause skin lesions and even death due to septicaemia in crocodiles.

78 FR 72710 - Notice of Intent To Repatriate Cultural Items: Denver Art Museum, Denver, CO

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2013-12-03

....R50000] Notice of Intent To Repatriate Cultural Items: Denver Art Museum, Denver, CO AGENCY: National Park Service, Interior. ACTION: Notice. SUMMARY: The Denver Art Museum, in consultation with the appropriate Indian tribes or Native Hawaiian organizations, has determined that the cultural items listed in...

Inference of Interactions in Cyanobacterial-Heterotrophic Co-Cultures via Transcriptome Sequencing

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Beliaev, Alex S.; Romine, Margaret F.; Serres, Margaret; Bernstein, Hans C.; Linggi, Bryan E.; Markillie, Lye Meng; Isern, Nancy G.; Chrisler, William B.; Kucek, Leo A.; Hill, Eric A.; Pinchuk, Grigoriy; Bryant, Donald A.; Wiley, H. S.; Fredrickson, Jim K.; Konopka, Allan

2014-04-29

We employed deep sequencing technology to identify transcriptional adaptation of the euryhaline unicellular cyanobacterium Synechococcus sp. PCC 7002 and the marine facultative aerobe Shewanella putrefaciens W3-18-1 to growth in a co-culture and infer the effect of carbon flux distributions on photoautotroph-heterotroph interactions. The overall transcriptome response of both organisms to co-cultivation was shaped by their respective physiologies and growth constraints. Carbon limitation resulted in the expansion of metabolic capacities which was manifested through the transcriptional upregulation of transport and catabolic pathways. While growth coupling occurred via lactate oxidation or secretion of photosynthetically fixed carbon, there was evidence of specific metabolic interactions between the two organisms. On one hand, the production and excretion of specific amino acids (methionine and alanine) by the cyanobacterium correlated with the putative downregulation of the corresponding biosynthetic machinery of Shewanella W3-18-1. On the other hand, the broad and consistent decrease of mRNA levels for many Fe-regulated Synechococcus 7002 genes during co-cultivation suggested increased Fe availability as well as more facile and energy-efficient mechanisms for Fe acquisition by the cyanobacterium. Furthermore, evidence pointed at potentially novel interactions between oxygenic photoautotrophs and heterotrophs related to the oxidative stress response as transcriptional patterns suggested that Synechococcus 7002 rather than Shewanella W3-18-1 provided scavenging functions for reactive oxygen species under co-culture conditions. This study provides an initial insight into the complexity of photoautotrophic-heterotrophic interactions and brings new perspectives of their role in the robustness and stability of the association.

Effectiveness of ambon banana stem juice as immunostimulatory against Aeromonas hydrophila infections in catfish Clarias gariepinus

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Qorie Astria; Sri Nuryati; Kukuh Nirmala; Alimuddin Alimuddin

2017-01-01

ABSTRACT Outbreaks of infectious diseases due to Aeromonas hydrophila in catfish can cause high death rates (80–100%). Fish disease control can be done using phytopharmaceutical to prevent or treat diseases of fish. One of the phytopharmaceutical that known to prevent the fish diseases is ambon banana stem Musa cavendishii var. dwarf Paxton. This study was conducted to test the effectiveness of catfish immersion using banana stem juice as an immunostimulant against bacterial infections A. hyd...

Streptococcus caviae sp. nov., isolated from guinea pig faecal samples.

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Palakawong Na Ayudthaya, Susakul; Hilderink, Loes J; Oost, John van der; Vos, Willem M de; Plugge, Caroline M

2017-05-01

A novel cellobiose-degrading and lactate-producing bacterium, strain Cavy grass 6T, was isolated from faecal samples of guinea pigs (Cavia porcellus). Cells of the strain were ovalshaped, non-motile, non-spore-forming, Gram-stain-positive and facultatively anaerobic. The strain gr at 25-40 °C (optimum 37 °C) and pH 4.5-9.5 (optimum 8.0). Phylogenetic analysis based on 16S rRNA gene sequences showed that strain Cavy grass 6T belongs to the genus Streptococcus with its closest relative being Streptococcus devriesei CCUG 47155T with only 96.5 % similarity. Comparing strain Cavy grass 6T and Streptococcus devriesei CCUG 47155T, average nucleotide identity and level of digital DNA-DNA hybridization dDDH were only 86.9 and 33.3 %, respectively. Housekeeping genes groEL and gyrA were different between strain Cavy grass 6T and other streptococci. The G+C content of strain Cavy grass 6T was 42.6±0.3 mol%. The major (>10 %) cellular fatty acids of strain Cavy grass 6T were C16:0, C20 : 1ω9c and summed feature 8 (C18 : 1ω7c and/or C18 : 1ω6c). Strain Cavy grass 6T ferment a range of plant mono- and disaccharides as well as polymeric carbohydrates, including cellobiose, dulcitol, d-glucose, maltose, raffinose, sucrose, l-sorbose, trehalose, inulin and dried grass extract, to lactate, formate, acetate and ethanol. Based on phylogenetic and physiological characteristics, Cavy grass 6T can be distinguished from other members of the genus Streptococcus. Therefore, a novel species of the genus Streptococcus, family Streptococcaceae, order Lactobacillales is proposed, Streptococcuscaviae sp. nov. (type strain Cavy grass 6T=TISTR 2371T=DSM 102819T).

Co-culture of Adult Mesenchymal Stem Cells and Nucleus Pulposus Cells in Bilaminar Pellets for Intervertebral Disc Regeneration.

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Allon, Aliza A; Schneider, Richard A; Lotz, Jeffrey C

2009-01-01

Our goal is to optimize stem cell-based tissue engineering strategies in the context of the intervertebral disc environment. We explored the benefits of co-culturing nucleus pulposus cells (NPC) and adult mesenchymal stem cells (MSC) using a novel spherical bilaminar pellet culture system where one cell type is enclosed in a sphere of the other cell type. Our 3D system provides a structure that exploits embryonic processes such as tissue induction and condensation. We observed a unique phenomenon: the budding of co-culture pellets and the formation of satellite pellets that separate from the main pellet. MSC and NPC co-culture pellets were formed with three different structural organizations. The first had random organization. The other two had bilaminar organization with either MSC inside and NPC outside or NPC inside and MSC outside. By 14 days, all co-culture pellets exhibited budding and spontaneously generated satellite pellets. The satellite pellets were composed of both cell types and, surprisingly, all had the same bilaminar organization with MSC on the inside and NPC on the outside. This organization was independent of the structure of the main pellet that the satellites stemmed from. The main pellets generated satellite pellets that spontaneously organized into a bilaminar structure. This implies that structural organization occurs naturally in this cell culture system and may be inherently favorable for cell-based tissue engineering strategies. The occurrence of budding and the organization of satellite pellets may have important implications for the use of co-culture pellets in cell-based therapies for disc regeneration. From a therapeutic point of view, the generation of satellite pellets may be a beneficial feature that would serve to spread donor cells throughout the host matrix and restore normal matrix composition in a sustainable way, ultimately renewing tissue function.

Co-cultures provide a new tool to probe communication between adult sensory neurons and urothelium.

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O'Mullane, Lauren M; Keast, Janet R; Osborne, Peregrine B

2013-08-01

Recent evidence suggests that the urothelium functions as a sensory transducer of chemical, mechanical or thermal stimuli and signals to nerve terminals and other cells in the bladder wall. The cellular and molecular basis of neuro-urothelial communication is not easily studied in the intact bladder. This led us to establish a method of co-culturing dorsal root ganglion sensory neurons and bladder urothelial cells. Sensory neurons and urothelial cells obtained from dorsal root ganglia and bladders dissected from adult female Sprague-Dawley® rats were isolated by enzyme treatment and mechanical dissociation. They were plated together or separately on collagen coated substrate and cultured in keratinocyte medium for 48 to 72 hours. Retrograde tracer labeling was performed to identify bladder afferents used for functional testing. Neurite growth and complexity in neurons co-cultured with urothelial cells was increased relative to that in neuronal monocultures. The growth promoting effect of urothelial cells was reduced by the tyrosine kinase inhibitor K252a but upstream inhibition of nerve growth factor signaling with TrkA-Fc had no effect. Fura-2 calcium imaging of urothelial cells showed responses to adenosine triphosphate (100 μM) and activation of TRPV4 (4α-PDD, 10 μM) but not TRPV1 (capsaicin, 1 μM), TRPV3 (farnesyl pyrophosphate, 1 μM) or TRPA1 (mustard oil, 100 μM). In contrast, co-cultured neurons were activated by all agonists except farnesyl pyrophosphate. Co-culturing provides a new methodology for investigating neuro-urothelial interactions in animal models of urological conditions. Results suggest that neuronal properties are maintained in the presence of urothelium and neurite growth is potentiated by a nerve growth factor independent mechanism. Copyright © 2013 American Urological Association Education and Research, Inc. Published by Elsevier Inc. All rights reserved.

Dynamics of bone marrow-derived endothelial progenitor cell/mesenchymal stem cell interaction in co-culture and its implications in angiogenesis

International Nuclear Information System (INIS)

Aguirre, A.; Planell, J.A.; Engel, E.

2010-01-01

Research highlights: → BM-EPCs and MSCs establish complex, self-organizing structures in co-culture. → Co-culture decreases proliferation by cellular self-regulatory mechanisms. → Co-cultured cells present an activated proangiogenic phenotype. → qRT-PCR and cluster analysis identify new target genes playing important roles. -- Abstract: Tissue engineering aims to regenerate tissues and organs by using cell and biomaterial-based approaches. One of the current challenges in the field is to promote proper vascularization in the implant to prevent cell death and promote host integration. Bone marrow endothelial progenitor cells (BM-EPCs) and mesenchymal stem cells (MSCs) are bone marrow resident stem cells widely employed for proangiogenic applications. In vivo, they are likely to interact frequently both in the bone marrow and at sites of injury. In this study, the physical and biochemical interactions between BM-EPCs and MSCs in an in vitro co-culture system were investigated to further clarify their roles in vascularization. BM-EPC/MSC co-cultures established close cell-cell contacts soon after seeding and self-assembled to form elongated structures at 3 days. Besides direct contact, cells also exhibited vesicle transport phenomena. When co-cultured in Matrigel, tube formation was greatly enhanced even in serum-starved, growth factor free medium. Both MSCs and BM-EPCs contributed to these tubes. However, cell proliferation was greatly reduced in co-culture and morphological differences were observed. Gene expression and cluster analysis for wide panel of angiogenesis-related transcripts demonstrated up-regulation of angiogenic markers but down-regulation of many other cytokines. These data suggest that cross-talk occurs in between BM-EPCs and MSCs through paracrine and direct cell contact mechanisms leading to modulation of the angiogenic response.

A New In Vitro Co-Culture Model Using Magnetic Force-Based Nanotechnology

NARCIS (Netherlands)

Takanari, Hiroki; Miwa, Keiko; Fu, Xian Ming; Nakai, Junichi; Ito, Akira; Ino, Kousuke; Honda, Hiroyuki; Tonomura, Wataru; Konishi, Satoshi; Opthof, Tobias; van der Heyden, Marcel A G; Kodama, Itsuo; Lee, Jong Kook

2016-01-01

Skeletal myoblast (SkMB) transplantation has been conducted as a therapeutic strategy for severe heart failure. However, arrhythmogenicity following transplantation remains unsolved. We developed an in vitro model of myoblast transplantation with “patterned” or “randomly-mixed” co-culture of SkMBs

Impact of sequential co-culture fermentations on flavour characters of Solaris wines

DEFF Research Database (Denmark)

Liu, Jing; Arneborg, Nils; Toldam-Andersen, Torben Bo

2017-01-01

-phenylethyl acetate. This co-culture fermentation also produced higher amounts of ethyl acetate and acetic acid, reducing the possibility of its use in winemaking. Three Metschnikowia strains, a M. chrysoperlae strain and two M. fructicola strains, gave a comparable production of volatile compounds...

Effects of CO 2 concentration and moisture content of sugar-free media on the tissue-cultured plantlets in a large growth chamber

Science.gov (United States)

Qu, Y. H.; Lin, C.; Zhou, W.; Li, Y.; Chen, B.; Chen, G. Q.

2009-01-01

The dynamic fluctuations of CO 2 concentration in the tissue culture growth chamber after transplantation of petunia, chrysanthemum and tomato plantlets were recorded with a real-time control system to determine the critical CO 2 concentration levels of 35 μl l -1 at which CO 2 enrichment is needed. The experimental data showed that the tissue-cultured plantlets of petunia, chrysanthemum and tomato had the same CO 2 concentration dynamics. The results indicated that CO 2 enrichment was proper on the second day after transplantation. Petunia plantlets were used to conduct experiments under PPFD of 80 μmol m -2 s -1, and CO 2 concentrations of 350 ± 50 μl l -1, 650 ± 50 μl l -1 and 950 ± 50 μl l -1 as well as medium moisture contents of 60%, 70% and 80%, with the result that plantlets grew better under CO 2 concentration of 650 ± 50 μl l -1 than under the other two concentrations with all the different media water contents. Three media water contents under the same CO 2 concentration produced plantlets with the same quality. The impacts of CO 2 concentrations on plantlets are more important than those of the media water contents. Sugar-free tissue culture, as compared with the conventional culture, showed that CO 2 enrichment to 350 ± 50 μl l -1 can promote the growth of the cultured plantlets. Sugar-free tissue culture produced healthy plantlets with thick roots, almost equivalent to the common plantlets.

Puerarin exhibits greater distribution and longer retention time in neurons than astrocytes in a co-cultured system

Directory of Open Access Journals (Sweden)

Shu-Yong Wei

2015-01-01

Full Text Available The phytoestrogen puerarin has been shown to protect neurons and astrocytes in the brain, and is therefore an attractive drug in the treatment of Alzheimer′s disease, Parkinson′s disease and cerebral ischemia. Whether puerarin exhibits the same biological processes in neurons and astrocytes in vitro has rarely been reported. In this study, cortical neurons and astrocytes of newborn Sprague-Dawley rats were separated, identified and co-cultured in a system based on Transwell membranes. The retention time and distribution of puerarin in each cell type was detected by fluorescence spectrophotometry and fluorescence microscope. The concentration of puerarin in both co-cultured and separately cultured neurons was greater than that of astrocytes. Puerarin concentration reached a maximum 20 minutes after it was added. At 60 minutes after its addition, a scant amount of drug was detected in astrocytes; however in both separately cultured and co-cultured neurons, the concentration of puerarin achieved a stable level of about 12.8 ng/mL. The results indicate that puerarin had a higher concentration and longer retention time in neurons than that observed in astrocytes.

White Mulberry (Morus alba Foliage Methanolic Extract Can Alleviate Aeromonas hydrophila Infection in African Catfish (Clarias gariepinus

Directory of Open Access Journals (Sweden)

Atefeh Sheikhlar

2014-01-01

Full Text Available Two experiments were simultaneously conducted with Morus alba (white mulberry foliage extract (MFE as a growth promoter and treatment of Aeromonas hydrophila infection in separate 60 and 30 days trail (Experiments 1 and 2, resp. in African catfish (Clarias gariepinus. In Experiment 1, four diets, control and control supplemented with 2, 5, or 7 g MFE/kg dry matter (DM of diet, were used. In Experiment 2, fish were intraperitoneally infected with Aeromonas hydrophila and fed the same diets as experiment 1 plus additional two diets with or without antibiotic. Results of experiment 1 showed that growth was unaffected by dietary levels of MFE. Treatments with the inclusion of MFE at the levels of 5 and 7 g/Kg DM had no mortality. Red blood cells (RBC, albumin, and total protein were all higher for the treatments fed MFE (5 and 7 g/Kg DM. Results of experiment 2 showed RBC, hemoglobin, hematocrit, globulin, albumin, and total protein improved with the increase in MFE in the infected fish. The dietary MFE at the level of 7 g/kg DM reduced mortality rate. In conclusion, MFE at the level of 7 g/kg DM could be a valuable dietary supplement to cure the infected fish.

White Mulberry (Morus alba) Foliage Methanolic Extract Can Alleviate Aeromonas hydrophila Infection in African Catfish (Clarias gariepinus)

Science.gov (United States)

Sheikhlar, Atefeh; Alimon, Abd Razk; Daud, Hassan; Saad, Chee R.; Webster, Carl D.; Meng, Goh Yong

2014-01-01

Two experiments were simultaneously conducted with Morus alba (white mulberry) foliage extract (MFE) as a growth promoter and treatment of Aeromonas hydrophila infection in separate 60 and 30 days trail (Experiments 1 and 2, resp.) in African catfish (Clarias gariepinus). In Experiment 1, four diets, control and control supplemented with 2, 5, or 7 g MFE/kg dry matter (DM) of diet, were used. In Experiment 2, fish were intraperitoneally infected with Aeromonas hydrophila and fed the same diets as experiment 1 plus additional two diets with or without antibiotic. Results of experiment 1 showed that growth was unaffected by dietary levels of MFE. Treatments with the inclusion of MFE at the levels of 5 and 7 g/Kg DM had no mortality. Red blood cells (RBC), albumin, and total protein were all higher for the treatments fed MFE (5 and 7 g/Kg DM). Results of experiment 2 showed RBC, hemoglobin, hematocrit, globulin, albumin, and total protein improved with the increase in MFE in the infected fish. The dietary MFE at the level of 7 g/kg DM reduced mortality rate. In conclusion, MFE at the level of 7 g/kg DM could be a valuable dietary supplement to cure the infected fish. PMID:25574488

White mulberry (Morus alba) foliage methanolic extract can alleviate Aeromonas hydrophila infection in African catfish (Clarias gariepinus).

Science.gov (United States)

Sheikhlar, Atefeh; Alimon, Abd Razk; Daud, Hassan; Saad, Chee R; Webster, Carl D; Meng, Goh Yong; Ebrahimi, Mahdi

2014-01-01

Two experiments were simultaneously conducted with Morus alba (white mulberry) foliage extract (MFE) as a growth promoter and treatment of Aeromonas hydrophila infection in separate 60 and 30 days trail (Experiments 1 and 2, resp.) in African catfish (Clarias gariepinus). In Experiment 1, four diets, control and control supplemented with 2, 5, or 7 g MFE/kg dry matter (DM) of diet, were used. In Experiment 2, fish were intraperitoneally infected with Aeromonas hydrophila and fed the same diets as experiment 1 plus additional two diets with or without antibiotic. Results of experiment 1 showed that growth was unaffected by dietary levels of MFE. Treatments with the inclusion of MFE at the levels of 5 and 7 g/Kg DM had no mortality. Red blood cells (RBC), albumin, and total protein were all higher for the treatments fed MFE (5 and 7 g/Kg DM). Results of experiment 2 showed RBC, hemoglobin, hematocrit, globulin, albumin, and total protein improved with the increase in MFE in the infected fish. The dietary MFE at the level of 7 g/kg DM reduced mortality rate. In conclusion, MFE at the level of 7 g/kg DM could be a valuable dietary supplement to cure the infected fish.

Melanization and Pathogenicity in the Insect, Tenebrio molitor, and the Crustacean, Pacifastacus leniusculus, by Aeromonas hydrophila AH-3

Science.gov (United States)

Noonin, Chadanat; Jiravanichpaisal, Pikul; Söderhäll, Irene; Merino, Susana; Tomás, Juan M.; Söderhäll, Kenneth

2010-01-01

Aeromonas hydrophila is the most common Aeromonas species causing infections in human and other animals such as amphibians, reptiles, fish and crustaceans. Pathogenesis of Aeromonas species have been reported to be associated with virulence factors such as lipopolysaccharides (LPS), bacterial toxins, bacterial secretion systems, flagella, and other surface molecules. Several mutant strains of A. hydrophila AH-3 were initially used to study their virulence in two animal species, Pacifastacus leniusculus (crayfish) and Tenebrio molitor larvae (mealworm). The AH-3 strains used in this study have mutations in genes involving the synthesis of flagella, LPS structures, secretion systems, and some other factors, which have been reported to be involved in A. hydrophila pathogenicity. Our study shows that the LPS (O-antigen and external core) is the most determinant A. hydrophila AH-3 virulence factor in both animals. Furthermore, we studied the immune responses of these hosts to infection of virulent or non-virulent strains of A. hydrophila AH-3. The AH-3 wild type (WT) containing the complete LPS core is highly virulent and this bacterium strongly stimulated the prophenoloxidase activating system resulting in melanization in both crayfish and mealworm. In contrast, the ΔwaaE mutant which has LPS without O-antigen and external core was non-virulent and lost ability to stimulate this system and melanization in these two animals. The high phenoloxidase activity found in WT infected crayfish appears to result from a low expression of pacifastin, a prophenoloxidase activating enzyme inhibitor, and this gene expression was not changed in the ΔwaaE mutant infected animal and consequently phenoloxidase activity was not altered as compared to non-infected animals. Therefore we show that the virulence factors of A. hydrophila are the same regardless whether an insect or a crustacean is infected and the O-antigen and external core is essential for activation of the proPO system

Melanization and pathogenicity in the insect, Tenebrio molitor, and the crustacean, Pacifastacus leniusculus, by Aeromonas hydrophila AH-3.

Directory of Open Access Journals (Sweden)

Chadanat Noonin

Full Text Available Aeromonas hydrophila is the most common Aeromonas species causing infections in human and other animals such as amphibians, reptiles, fish and crustaceans. Pathogenesis of Aeromonas species have been reported to be associated with virulence factors such as lipopolysaccharides (LPS, bacterial toxins, bacterial secretion systems, flagella, and other surface molecules. Several mutant strains of A. hydrophila AH-3 were initially used to study their virulence in two animal species, Pacifastacus leniusculus (crayfish and Tenebrio molitor larvae (mealworm. The AH-3 strains used in this study have mutations in genes involving the synthesis of flagella, LPS structures, secretion systems, and some other factors, which have been reported to be involved in A. hydrophila pathogenicity. Our study shows that the LPS (O-antigen and external core is the most determinant A. hydrophila AH-3 virulence factor in both animals. Furthermore, we studied the immune responses of these hosts to infection of virulent or non-virulent strains of A. hydrophila AH-3. The AH-3 wild type (WT containing the complete LPS core is highly virulent and this bacterium strongly stimulated the prophenoloxidase activating system resulting in melanization in both crayfish and mealworm. In contrast, the ΔwaaE mutant which has LPS without O-antigen and external core was non-virulent and lost ability to stimulate this system and melanization in these two animals. The high phenoloxidase activity found in WT infected crayfish appears to result from a low expression of pacifastin, a prophenoloxidase activating enzyme inhibitor, and this gene expression was not changed in the ΔwaaE mutant infected animal and consequently phenoloxidase activity was not altered as compared to non-infected animals. Therefore we show that the virulence factors of A. hydrophila are the same regardless whether an insect or a crustacean is infected and the O-antigen and external core is essential for activation of the

Human neuron-astrocyte 3D co-culture-based assay for evaluation of neuroprotective compounds.

Science.gov (United States)

Terrasso, Ana Paula; Silva, Ana Carina; Filipe, Augusto; Pedroso, Pedro; Ferreira, Ana Lúcia; Alves, Paula Marques; Brito, Catarina

Central nervous system drug development has registered high attrition rates, mainly due to the lack of efficacy of drug candidates, highlighting the low reliability of the models used in early-stage drug development and the need for new in vitro human cell-based models and assays to accurately identify and validate drug candidates. 3D human cell models can include different tissue cell types and represent the spatiotemporal context of the original tissue (co-cultures), allowing the establishment of biologically-relevant cell-cell and cell-extracellular matrix interactions. Nevertheless, exploitation of these 3D models for neuroprotection assessment has been limited due to the lack of data to validate such 3D co-culture approaches. In this work we combined a 3D human neuron-astrocyte co-culture with a cell viability endpoint for the implementation of a novel in vitro neuroprotection assay, over an oxidative insult. Neuroprotection assay robustness and specificity, and the applicability of Presto Blue, MTT and CytoTox-Glo viability assays to the 3D co-culture were evaluated. Presto Blue was the adequate endpoint as it is non-destructive and is a simpler and reliable assay. Semi-automation of the cell viability endpoint was performed, indicating that the assay setup is amenable to be transferred to automated screening platforms. Finally, the neuroprotection assay setup was applied to a series of 36 test compounds and several candidates with higher neuroprotective effect than the positive control, Idebenone, were identified. The robustness and simplicity of the implemented neuroprotection assay with the cell viability endpoint enables the use of more complex and reliable 3D in vitro cell models to identify and validate drug candidates. Copyright © 2016 Elsevier Inc. All rights reserved.

A long-term three dimensional liver co-culture system for improved prediction of clinically relevant drug-induced hepatotoxicity

International Nuclear Information System (INIS)

Kostadinova, Radina; Boess, Franziska; Applegate, Dawn; Suter, Laura; Weiser, Thomas; Singer, Thomas; Naughton, Brian; Roth, Adrian

2013-01-01

Drug-induced liver injury (DILI) is the major cause for liver failure and post-marketing drug withdrawals. Due to species-specific differences in hepatocellular function, animal experiments to assess potential liabilities of drug candidates can predict hepatotoxicity in humans only to a certain extent. In addition to animal experimentation, primary hepatocytes from rat or human are widely used for pre-clinical safety assessment. However, as many toxic responses in vivo are mediated by a complex interplay among different cell types and often require chronic drug exposures, the predictive performance of hepatocytes is very limited. Here, we established and characterized human and rat in vitro three-dimensional (3D) liver co-culture systems containing primary parenchymal and non-parenchymal hepatic cells. Our data demonstrate that cells cultured on a 3D scaffold have a preserved composition of hepatocytes, stellate, Kupffer and endothelial cells and maintain liver function for up to 3 months, as measured by the production of albumin, fibrinogen, transferrin and urea. Additionally, 3D liver co-cultures maintain cytochrome P450 inducibility, form bile canaliculi-like structures and respond to inflammatory stimuli. Upon incubation with selected hepatotoxicants including drugs which have been shown to induce idiosyncratic toxicity, we demonstrated that this model better detected in vivo drug-induced toxicity, including species-specific drug effects, when compared to monolayer hepatocyte cultures. In conclusion, our results underline the importance of more complex and long lasting in vitro cell culture models that contain all liver cell types and allow repeated drug-treatments for detection of in vivo-relevant adverse drug effects. - Highlights: ► 3D liver co-cultures maintain liver specific functions for up to three months. ► Activities of Cytochrome P450s remain drug- inducible accross three months. ► 3D liver co-cultures recapitulate drug-induced liver toxicity

A long-term three dimensional liver co-culture system for improved prediction of clinically relevant drug-induced hepatotoxicity

Energy Technology Data Exchange (ETDEWEB)

Kostadinova, Radina; Boess, Franziska [Non-Clinical Safety, Hoffmann-La Roche Ltd, Grenzacherstrasse 124, Building 73 / Room 117b, 4070 Basel (Switzerland); Applegate, Dawn [RegeneMed, 9855 Towne Centre Drive Suite 200, San Diego, CA 92121 (United States); Suter, Laura; Weiser, Thomas; Singer, Thomas [Non-Clinical Safety, Hoffmann-La Roche Ltd, Grenzacherstrasse 124, Building 73 / Room 117b, 4070 Basel (Switzerland); Naughton, Brian [RegeneMed, 9855 Towne Centre Drive Suite 200, San Diego, CA 92121 (United States); Roth, Adrian, E-mail: adrian_b.roth@roche.com [Non-Clinical Safety, Hoffmann-La Roche Ltd, Grenzacherstrasse 124, Building 73 / Room 117b, 4070 Basel (Switzerland)

2013-04-01

Drug-induced liver injury (DILI) is the major cause for liver failure and post-marketing drug withdrawals. Due to species-specific differences in hepatocellular function, animal experiments to assess potential liabilities of drug candidates can predict hepatotoxicity in humans only to a certain extent. In addition to animal experimentation, primary hepatocytes from rat or human are widely used for pre-clinical safety assessment. However, as many toxic responses in vivo are mediated by a complex interplay among different cell types and often require chronic drug exposures, the predictive performance of hepatocytes is very limited. Here, we established and characterized human and rat in vitro three-dimensional (3D) liver co-culture systems containing primary parenchymal and non-parenchymal hepatic cells. Our data demonstrate that cells cultured on a 3D scaffold have a preserved composition of hepatocytes, stellate, Kupffer and endothelial cells and maintain liver function for up to 3 months, as measured by the production of albumin, fibrinogen, transferrin and urea. Additionally, 3D liver co-cultures maintain cytochrome P450 inducibility, form bile canaliculi-like structures and respond to inflammatory stimuli. Upon incubation with selected hepatotoxicants including drugs which have been shown to induce idiosyncratic toxicity, we demonstrated that this model better detected in vivo drug-induced toxicity, including species-specific drug effects, when compared to monolayer hepatocyte cultures. In conclusion, our results underline the importance of more complex and long lasting in vitro cell culture models that contain all liver cell types and allow repeated drug-treatments for detection of in vivo-relevant adverse drug effects. - Highlights: ► 3D liver co-cultures maintain liver specific functions for up to three months. ► Activities of Cytochrome P450s remain drug- inducible accross three months. ► 3D liver co-cultures recapitulate drug-induced liver toxicity

In vitro differentiation of bone marrow stromal cells into neurons and glial cells and differential protein expression in a two-compartment bone marrow stromal cell/neuron co-culture system.

Science.gov (United States)

Qi, Xu; Shao, Ming; Peng, Haisheng; Bi, Zhenggang; Su, Zhiqiang; Li, Hulun

2010-07-01

This study was performed to establish a bone marrow stromal cell (BMSC)/neuron two-compartment co-culture model in which differentiation of BMSCs into neurons could occur without direct contact between the two cell types, and to investigate protein expression changes during differentiation of this entirely BMSC-derived population. Cultured BMSCs isolated from Wistar rats were divided into three groups: BMSC culture, BMSC/neuron co-culture and BMSC/neuron two-compartment co-culture. Cells were examined for neuron-specific enolase (NSE) and glial fibrillary acidic protein (GFAP) expression. The electrophysiological behavior of the BMSCs was examined using patch clamping. Proteins that had significantly different expression levels in BMSCs cultured alone and co-cultured with neurons were studied using a protein chip-mass spectroscopy technique. Expression of NSE and GFAP were significantly higher in co-culture cells than in two-compartment co-culture cells, and significantly higher in both co-culture groups than in BMSCs cultured alone. Five proteins showed significant changes in expression during differentiation: TIP39_RAT and CALC_RAT underwent increases, and INSL6_RAT, PNOC_RAT and PCSK1_RAT underwent decreases in expression. We conclude that BMSCs can differentiate into neurons during both contact co-culture with neurons and two-compartment co-culture with neurons. The rate at which BMSCs differentiated into neurons was higher in contact co-culture than in non-contact co-culture.

Production systems, diversity and richness of cavy culture in ...

African Journals Online (AJOL)

In order to assess existing cavy (Cavia porcellus) production systems of Cameroon western highlands and rain forest agro-ecological zones, a household ... for a well designed and comprehensive national breeding program for cavies, and increased capacity building of farmers to address mortality rates and health issues.

Porphyromonas gingivalis displays a competitive advantage over Aggregatibacter actinomycetemcomitans in co-cultured biofilm.

Science.gov (United States)

Takasaki, K; Fujise, O; Miura, M; Hamachi, T; Maeda, K

2013-06-01

Biofilm formation occurs through the events of cooperative growth and competitive survival among multiple species. Porphyromonas gingivalis and Aggregatibacter actinomycetemcomitans are important periodontal pathogens. The aim of this study was to demonstrate competitive or cooperative interactions between these two species in co-cultured biofilm. P. gingivalis strains and gingipain mutants were cultured with or without A. actinomycetemcomitans. Biofilms formed on glass surfaces were analyzed by crystal violet staining and colony counting. Preformed A. actinomycetemcomitans biofilms were treated with P. gingivalis culture supernatants. Growth and proteolytic activities of gingipains were also determined. Monocultured P. gingivalis strains exhibited a range of biofilm-formation abilities and proteolytic activities. The ATCC33277 strain, noted for its high biofilm-formation ability and proteolytic activity, was found to be dominant in biofilm co-cultured with A. actinomycetemcomitans. In a time-resolved assay, A. actinomycetemcomitans was primarily the dominant colonizer on a glass surface and subsequently detached in the presence of increasing numbers of ATCC33277. Detachment of preformed A. actinomycetemcomitans biofilm was observed by incubation with culture supernatants from highly proteolytic strains. These results suggest that P. gingivalis possesses a competitive advantage over A. actinomycetemcomitans. As the required biofilm-formation abilities and proteolytic activities vary among P. gingivalis strains, the diversity of the competitive advantage is likely to affect disease recurrence during periodontal maintenance. © 2012 John Wiley & Sons A/S. Published by John Wiley & Sons Ltd.

Dynamic flux balance modeling of microbial co-cultures for efficient batch fermentation of glucose and xylose mixtures.

Science.gov (United States)

Hanly, Timothy J; Henson, Michael A

2011-02-01

Sequential uptake of pentose and hexose sugars that compose lignocellulosic biomass limits the ability of pure microbial cultures to efficiently produce value-added bioproducts. In this work, we used dynamic flux balance modeling to examine the capability of mixed cultures of substrate-selective microbes to improve the utilization of glucose/xylose mixtures and to convert these mixed substrates into products. Co-culture simulations of Escherichia coli strains ALS1008 and ZSC113, engineered for glucose and xylose only uptake respectively, indicated that improvements in batch substrate consumption observed in previous experimental studies resulted primarily from an increase in ZSC113 xylose uptake relative to wild-type E. coli. The E. coli strain ZSC113 engineered for the elimination of glucose uptake was computationally co-cultured with wild-type Saccharomyces cerevisiae, which can only metabolize glucose, to determine if the co-culture was capable of enhanced ethanol production compared to pure cultures of wild-type E. coli and the S. cerevisiae strain RWB218 engineered for combined glucose and xylose uptake. Under the simplifying assumption that both microbes grow optimally under common environmental conditions, optimization of the strain inoculum and the aerobic to anaerobic switching time produced an almost twofold increase in ethanol productivity over the pure cultures. To examine the effect of reduced strain growth rates at non-optimal pH and temperature values, a break even analysis was performed to determine possible reductions in individual strain substrate uptake rates that resulted in the same predicted ethanol productivity as the best pure culture. © 2010 Wiley Periodicals, Inc.

Cultural Analysis - towards cross-cultural understanding

DEFF Research Database (Denmark)

Gullestrup, Hans

The book considers intercultural understanding and co-action, partly by means of general insight into concept of culture and the dimensions which bring about cultural differences, and partly as a methodology to analyse a certain culture - whether one's own or others'. This leads towards an unders......The book considers intercultural understanding and co-action, partly by means of general insight into concept of culture and the dimensions which bring about cultural differences, and partly as a methodology to analyse a certain culture - whether one's own or others'. This leads towards...... a theoretical/abstract proposal for cultural understanding. The second part presents a theoretical/abstract proposal for under-standing intercultural plurality and complexity. The third part provides an empirical model for the analysis of intercultural co-action. Finally, the fourth part present and discusses...

Triple co-culture cell model as an in vitro model for oral particulate vaccine systems

DEFF Research Database (Denmark)

Nielsen, Line Hagner; De Rossi, C.; Lehr, C.-M.

the immunostimulatory ability of particulate vaccine formulations designed for oral delivery. Levels of cytokine production in response to vaccine administration were measured following particulate vaccine administration, as an indication of dendritic cell and macrophage activation. Precursors of cubosomes containing......; this was not observed with ovalbumin and blank solution. An example of the results is shown in Figure 2 for IL-17A. An established co-culture of Caco-2, THP-1 and MUTZ-3 cells showed promise as an in vitro model for testing of oral vaccine formulations. Mobility of co-culture immune cells as well as cytokine production...... with particle formulations. This was not the case when incubating with ovalbumin solution or blank. The ELISA screening assay showed production of a wide range of cytokines following culture incubation with cubosomes (with and without ovalbumin) and LPS solutions, indicative of a stimulatory effect...

Education for a Culture of Peace and Co-Existence

Directory of Open Access Journals (Sweden)

Silvia Guetta

2016-07-01

Full Text Available Education is a key and fundamental tool required to achieve social change, especially regarding social cohesion and co-existence. Education affects the most critical issues facing humanity, including the proliferation of various forms of violence, environmental degradation, and annihilation of cultures. I argue that the impact of education is equal to that of economics, politics, and technological advances. Therefore, if we wish to see change, educational reform has to be designed to operate in a global-human context, empowering and allowing people to achieve their potential. It should instil in individuals and groups respect for others with whom they interact, in a global as well as local sense. The article draws on the theories of Edgar Morin, Reuven Feuerstein, and Jerome Bruner, and their contributions to the development of educational approaches that encourage a culture of responsible, participatory, and creative coexistence. .

Cultural, social and intrapersonal factors associated with clusters of co-occurring health-related behaviours among adolescents

NARCIS (Netherlands)

Klein Velderman, Mariska; Dusseldorp, Elise; van Nieuwenhuijzen, Maroesjka; Junger, Marianne; Paulussen, Theo G. W. M.; Reijneveld, Sijmen A.

BACKGROUND: Adverse health-related behaviours (HRBs) have been shown to co-occur in adolescents. Evidence lacks on factors associated with these co-occurring HRBs. The Theory of Triadic Influence (TTI) offers a route to categorize these determinants according to type (social, cultural and

Slight changes in the mechanical stimulation affects osteoblast- and osteoclast-like cells in co-culture.

Science.gov (United States)

Kadow-Romacker, Anke; Duda, Georg N; Bormann, Nicole; Schmidmaier, Gerhard; Wildemann, Britt

2013-12-01

Osteoblast- and osteoclast-like cells are responsible for coordinated bone maintenance, illustrated by a balanced formation and resorption. Both parameters appear to be influenced by mechanical constrains acting on each of these cell types individually. We hypothesized that the interactions between both cell types are also influenced by mechanical stimulation. Co-cultures of osteoblast- and osteoclast-like cells were stimulated with 1,100 µstrain, 0.1 or 0.3 Hz for 1-5 min/day over 5 days. Two different setups depending on the differentiation of the osteoclast-like cells were used: i) differentiation assay for the fusion of pre-osteoclasts to osteoclasts, ii) resorption assay to determine the activity level of osteoclast-like cells. In the differentiation assay (co-culture of osteoblasts with unfused osteoclast precursor cells) the mechanical stimulation resulted in a significant decrease of collagen-1 and osteocalcin produced by osteoblast-like cells. Significantly more TRAP-iso5b was measured after stimulation for 3 min with 0.1 Hz, indicating enhanced osteoclastogenesis. In the resorption assay (co-culture of osteoblasts with fused osteoclasts) the stimulation for 3 min with 0.3 Hz significantly increased the resorption activity of osteoclasts measured by the pit formation and the collagen resorption. The same mechanical stimulation resulted in an increased collagen-1 production by the osteoblast-like cells. The ratio of RANKL/OPG was not different between the groups. These findings demonstrate that already small changes in duration or frequency of mechanical stimulation had significant consequences for the behavior of osteoblast- and osteoclast-like cells in co-culture, which partially depend on the differentiation status of the osteoclast-like cells.

Culture as Mediator: Co-Regulation, Self-Regulation, and Middle School Mathematics Achievement

Science.gov (United States)

Hinnant-Crawford, Brandi Nicole; Faison, Morgan Z.; Chang, Mei-Lin

2016-01-01

Purpose: Self-regulation is defined as strategic, metacognitive behavior, motivation and cognition aimed at a goal (Zimmmerman and Schunk, 2011). Co-regulation, arguably more aligned with norms in communal cultures, is the process of learners sharing "a common problem-solving plane" through which self-regulatory strategies are learned…

Skin diseases in companion guinea pigs (Cavia porcellus): a retrospective study of 293 cases seen at the Veterinary Medical Teaching Hospital, University of California at Davis (1990-2015).

Science.gov (United States)

White, Stephen D; Guzman, David Sanchez-Migallon; Paul-Murphy, Joanne; Hawkins, Michelle G

2016-10-01

Guinea pigs (Cavia porcellus) are popular companion animals with reported skin diseases, but most reports are accessed from textbooks or review articles. To document skin diseases and their prevalence in companion guinea pigs in northern California, USA, and to investigate predilections for the most common conditions over a 25 year period. Two hundred and ninety three guinea pigs from the hospital population met inclusion criteria. A retrospective study was performed by searching computerized medical records seen at the Veterinary Medical Teaching Hospital (VMTH), School of Veterinary Medicine, University of California from 1 January 1990 to 31 July 2015 using key words relevant to dermatology. Of the 580 guinea pigs seen at the VMTH, 50% (293) had skin disease. Of the 293 cases, 154 (52%) presented for nondermatological reasons. Guinea pigs with skin disease were significantly older than those without skin disease (P = 0.0002); females with skin disease were more likely to have cystic ovaries (P = 0.0203), although these were not always associated with alopecia. Pododermatitis and infestation with Trixacarus caviae or lice were the most common skin diseases. Ivermectin or selamectin was used for ectoparasite treatment. Abscesses unassociated with pododermatitis were the most common nodules. Benign follicular tumours were the most common neoplasm. Despite the frequent mention of dermatophytosis in the veterinary literature, only two cases of Trichophyton mentagrophytes were diagnosed. Cutaneous conditions in companion guinea pigs in the USA are common. Clinicians should include a dermatological examination when examining these rodents regardless of the reason for presentation. © 2016 ESVD and ACVD.

Co-culture with human synovium-derived mesenchymal stem cells inhibits inflammatory activity and increases cell proliferation of sodium nitroprusside-stimulated chondrocytes

International Nuclear Information System (INIS)

Ryu, Jae-Sung; Jung, Yeon-Hwa; Cho, Mi-Young; Yeo, Jee Eun; Choi, Yun-Jin; Kim, Yong Il; Koh, Yong-Gon

2014-01-01

Highlights: • Co-culture of hSDMSCs with SNP-stimulated chondrocytes improves anti-inflammation. • Co-culture system produces IGF-1. • Co-culture system suppresses inflammatory genes expression. • Co-culture system improves cell proliferation. • Exogenous IGF-1 inhibits inflammatory activity in SNP-stimulated chondrocytes. - Abstract: Rheumatoid arthritis (RA) and osteoarthritis (OA) are primarily chronic inflammatory diseases. Mesenchymal stem cells (MSCs) have the ability to differentiate into cells of the mesodermal lineage, and to regulate immunomodulatory activity. Specifically, MSCs have been shown to secrete insulin-like growth factor 1 (IGF-1). The purpose of the present study was to examine the inhibitory effects on inflammatory activity from a co-culture of human synovium-derived mesenchymal stem cells (hSDMSCs) and sodium nitroprusside (SNP)-stimulated chondrocytes. First, chondrocytes were treated with SNP to generate an in vitro model of RA or OA. Next, the co-culture of hSDMSCs with SNP-stimulated chondrocytes reduced inflammatory cytokine secretion, inhibited expression of inflammation activity-related genes, generated IGF-1 secretion, and increased the chondrocyte proliferation rate. To evaluate the effect of IGF-1 on inhibition of inflammation, chondrocytes pre-treated with IGF-1 were treated with SNP, and then the production of inflammatory cytokines was analyzed. Treatment with IGF-1 was shown to significantly reduce inflammatory cytokine secretion in SNP-stimulated chondrocytes. Our results suggest that hSDMSCs offer a new strategy to promote cell-based cartilage regeneration in RA or OA

Co-culture with human synovium-derived mesenchymal stem cells inhibits inflammatory activity and increases cell proliferation of sodium nitroprusside-stimulated chondrocytes

Energy Technology Data Exchange (ETDEWEB)

Ryu, Jae-Sung; Jung, Yeon-Hwa; Cho, Mi-Young; Yeo, Jee Eun; Choi, Yun-Jin; Kim, Yong Il; Koh, Yong-Gon, E-mail: yonseranglab@daum.net

2014-05-16

Highlights: • Co-culture of hSDMSCs with SNP-stimulated chondrocytes improves anti-inflammation. • Co-culture system produces IGF-1. • Co-culture system suppresses inflammatory genes expression. • Co-culture system improves cell proliferation. • Exogenous IGF-1 inhibits inflammatory activity in SNP-stimulated chondrocytes. - Abstract: Rheumatoid arthritis (RA) and osteoarthritis (OA) are primarily chronic inflammatory diseases. Mesenchymal stem cells (MSCs) have the ability to differentiate into cells of the mesodermal lineage, and to regulate immunomodulatory activity. Specifically, MSCs have been shown to secrete insulin-like growth factor 1 (IGF-1). The purpose of the present study was to examine the inhibitory effects on inflammatory activity from a co-culture of human synovium-derived mesenchymal stem cells (hSDMSCs) and sodium nitroprusside (SNP)-stimulated chondrocytes. First, chondrocytes were treated with SNP to generate an in vitro model of RA or OA. Next, the co-culture of hSDMSCs with SNP-stimulated chondrocytes reduced inflammatory cytokine secretion, inhibited expression of inflammation activity-related genes, generated IGF-1 secretion, and increased the chondrocyte proliferation rate. To evaluate the effect of IGF-1 on inhibition of inflammation, chondrocytes pre-treated with IGF-1 were treated with SNP, and then the production of inflammatory cytokines was analyzed. Treatment with IGF-1 was shown to significantly reduce inflammatory cytokine secretion in SNP-stimulated chondrocytes. Our results suggest that hSDMSCs offer a new strategy to promote cell-based cartilage regeneration in RA or OA.

Effect of chest wall radiotherapy in different manners using tissue equivalent bolus on skin and lung of cavia cobayas

International Nuclear Information System (INIS)

Huang Wei; Qu Yaqin; Song Xiangfu; Liu Shixin; Jia Xiaojing; Guo He; Yang Lei

2009-01-01

Objective: To probe the influence of electron beam radiotherapy in different manners using different tissue equivalent boluses on skin and lung. Methods: Adult female cavia cobayas were randomly divided into four groups as control group, half-time with bolus group, half-time with bolus group and without bolus group. Acute-irradiation animal models were established using electron beam in different manners with or without 0.5 cm tissue equivalent bolus. Pathological changes in lung, hair vesicle and fibroblast cell count were analyzed 40 clays after irradiation. Results: The radiation dermatitis in the group with bolus was slighter than that of the group without bolus, but the radiation pneumonia was reverse. With bolus, the radiation dermatitis of haft-time group was slighter than that of full-time group. The injury repair of half-time group was more active than full-time group. Conclusions: The treatment of haft-time bolus could protect lung without serious skin complications. (authors)

Female novelty and the courtship behavior of male guinea pigs (Cavia porcellus

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Cohn D.W.H.

2004-01-01

Full Text Available In several rodent species, an increase or recovery of sexual behavior can be observed when sexually satiated males are placed in contact with a novel mate. In order to assess the influence of female novelty on the courtship behavior of guinea pigs (Cavia porcellus, four adult males were observed during four daily 15-min sessions while interacting with the same pregnant female (same-female sessions. A new female was presented during the fifth session (switched-female session. The duration of behavioral categories was obtained from videotape records using an observational software. From the first to the second session, all males decreased the time allocated to investigating (sniffing and licking, following, and mounting the female, and that response did not recover by the end of the same-female sessions. No similar decreasing tendencies were detected in the circling or rumba categories. A marked increase of investigating occurred in all males from the last same-female session (8.1, 11.9, 15.1 and 17.3 percent session time to the switched-female one (16.4, 18.4, 37.1 and 28.9 percent session time, respectively. Increases in following and circling were recorded in three of four males, and full-blown recovery of mounting in one male. No consistent changes in the females' responses to males (following or attacking were observed throughout testing. These results are consistent with the hypothesis that guinea pig males recognize individual females and that courtship responses may suffer a habituation/recovery process controlled by mate novelty.

Cultural, social and intrapersonal factors associated with clusters of co-occurring health-related behaviours among adolescents

NARCIS (Netherlands)

Klein Velderman, M.; Dusseldorp, E.; Nieuwenhuijzen, M. van; Paulussen, T.W.G.M.; Junger, M.; Reijneveld, S.A.

2015-01-01

Adverse health-related behaviours (HRBs) have been shown to co-occur in adolescents. Evidence lacks on factors associated with these co-occurring HRBs. The Theory of Triadic Influence (TTI) offers a route to categorize these determinants according to type (social, cultural and intrapersonal) and

Frozen White-Leg Shrimp (Litopenaeus vannamei) in Korean Markets as a Source of Aeromonas spp. Harboring Antibiotic and Heavy Metal Resistance Genes.

Science.gov (United States)

De Silva, Benthotage C J; Hossain, Sabrina; Dahanayake, Pasan S; Heo, Gang-Joon

2018-05-24

As the most consumed shrimp variety, white-leg shrimp (Litopenaeus vannamei) owns a high market demand in Korea. This study sought to screen the frozen white-leg shrimp for Aeromonas spp. harboring antimicrobial and heavy metal resistance characteristics. A total of 44 Aeromonas spp. strains were isolated and tested for antibiotic susceptibility and heavy metal tolerance followed by PCR-based detection of resistance genes and integrons. It was observed that resistance to ampicillin and oxacillin was 100% among isolates. Besides, 95%, 89%, 86%, 80%, 66%, and 43% of the isolates were resistant to nalidixic acid, tetracycline, cephalothin, streptomycin, trimethoprim-sulfamethoxazole, and imipenem, respectively, and less resistance to other antibiotics was also observed. Cr resistance was the highest (91%) among five heavy metals tested, whereas 57%, 32%, 20%, and 9% of the isolates were tolerant to Cu, Pb, Cd, and Hg, respectively. The PCR assays implied the presence of qnrB, qnrS, tetA, tetE, aac(6')-Ib, and aphAI-IAB, and intI1 genes among 80%, 77%, 18%, 30%, 9%, 0.25%, and 82% of the isolates, respectively. There were 35 (80%) integron 1-positive isolates harboring qacE2, dfrA1, orfC, orfD, aadB, catB3, oxa-10, and aadA1 genes in varying combinations. In addition, heavy metal resistance genes, CopA, merA, and CzcA were positive in 9%, 7%, and 27% of the isolates, respectively. According to these outcomes, the frozen white-leg shrimp in Korean markets can be suggested as a source of multidrug and heavy metal-resistant Aeromonas spp. that carries genetic determinants.

In vitro antimicrobial activity of ethanolic extracts obtained from Ficus spp. leaves against the fish pathogen Aeromonas hydrophila

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Tkachenko Halyna

2016-12-01

Full Text Available The main goal of this study was to determine in vitro antimicrobial activity of ethanolic extracts obtained from the leaves of various Ficus species against Aeromonas hydrophila isolated locally from infected rainbow trout (Oncorhynchus mykiss Walbaum with the aim of providing scientific rationale for the use of the plant in the treatment of bacterial infections induced by Aeromonas spp. in fish. Antimicrobial susceptibility testing was done on Muller-Hinton agar with the disc diffusion method. In the present study, most ethanolic extracts proved effective against the A. hydrophila tested, with 10-12 mm inhibition zones observed. A. hydrophila demonstrated the highest susceptibility to F. pumila. Among various species of Ficus with moderate activity against A. hydrophila, the highest antibacterial activities were noted for F. benghalensis, F. benjamina, F. deltoidea, F. hispida, and F. lyrata. Thus, Ficus can be used as a natural antiseptic and antimicrobial agent in veterinary practice. Further investigations need to be conducted to isolate and identify the bioactive compounds that can then be subjected to detailed pharmacological studies and the development of clinical applications. The alarming rate of increasing resistance in bacterial pathogens in aquaculture environments means that medicinal plants with antibacterial properties are very important as natural resources of new active compounds.

Contacting co-culture of human retinal microvascular endothelial cells alters barrier function of human embryonic stem cell derived retinal pigment epithelial cells.

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Skottman, H; Muranen, J; Lähdekorpi, H; Pajula, E; Mäkelä, K; Koivusalo, L; Koistinen, A; Uusitalo, H; Kaarniranta, K; Juuti-Uusitalo, K

2017-10-01

Here we evaluated the effects of human retinal microvascular endothelial cells (hREC) on mature human embryonic stem cell (hESC) derived retinal pigment epithelial (RPE) cells. The hESC-RPE cells (Regea08/017, Regea08/023 or Regea11/013) and hREC (ACBRI 181) were co-cultured on opposite sides of transparent membranes for up to six weeks. Thereafter barrier function, small molecule permeability, localization of RPE and endothelial cell marker proteins, cellular fine structure, and growth factor secretion of were evaluated. After co-culture, the RPE specific CRALBP and endothelial cell specific von Willebrand factor were appropriately localized. In addition, the general morphology, pigmentation, and fine structure of hESC-RPE cells were unaffected. Co-culture increased the barrier function of hESC-RPE cells, detected both with TEER measurements and cumulative permeability of FD4 - although the differences varied among the cell lines. Co-culturing significantly altered VEGF and PEDF secretion, but again the differences were cell line specific. The results of this study showed that co-culture with hREC affects hESC-RPE functionality. In addition, co-culture revealed drastic cell line specific differences, most notably in growth factor secretion. This model has the potential to be used as an in vitro outer blood-retinal barrier model for drug permeability testing. Copyright © 2017 The Authors. Published by Elsevier Inc. All rights reserved.

An improved in vitro blood-brain barrier model: rat brain endothelial cells co-cultured with astrocytes.

Science.gov (United States)

Abbott, N Joan; Dolman, Diana E M; Drndarski, Svetlana; Fredriksson, Sarah M

2012-01-01

In vitro blood-brain barrier (BBB) models using primary cultured brain endothelial cells are important for establishing cellular and molecular mechanisms of BBB function. Co-culturing with BBB-associated cells especially astrocytes to mimic more closely the in vivo condition leads to upregulation of the BBB phenotype in the brain endothelial cells. Rat brain endothelial cells (RBECs) are a valuable tool allowing ready comparison with in vivo studies in rodents; however, it has been difficult to obtain pure brain endothelial cells, and few models achieve a transendothelial electrical resistance (TEER, measure of tight junction efficacy) of >200 Ω cm(2), i.e. the models are still relatively leaky. Here, we describe methods for preparing high purity RBECs and neonatal rat astrocytes, and a co-culture method that generates a robust, stable BBB model that can achieve TEER >600 Ω cm(2). The method is based on >20 years experience with RBEC culture, together with recent improvements to kill contaminating cells and encourage BBB differentiation.Astrocytes are isolated by mechanical dissection and cell straining and are frozen for later co-culture. RBECs are isolated from 3-month-old rat cortices. The brains are cleaned of meninges and white matter and enzymatically and mechanically dissociated. Thereafter, the tissue homogenate is centrifuged in bovine serum albumin to separate vessel fragments from other cells that stick to the myelin plug. The vessel fragments undergo a second enzyme digestion to separate pericytes from vessels and break down vessels into shorter segments, after which a Percoll gradient is used to separate capillaries from venules, arterioles, and single cells. To kill remaining contaminating cells such as pericytes, the capillary fragments are plated in puromycin-containing medium and RBECs grown to 50-60% confluence. They are then passaged onto filters for co-culture with astrocytes grown in the bottom of the wells. The whole procedure takes ∼2

Effect of Cyclic Dynamic Compressive Loading on Chondrocytes and Adipose-Derived Stem Cells Co-Cultured in Highly Elastic Cryogel Scaffolds

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Chih-Hao Chen

2018-01-01

Full Text Available In this study, we first used gelatin/chondroitin-6-sulfate/hyaluronan/chitosan highly elastic cryogels, which showed total recovery from large strains during repeated compression cycles, as 3D scaffolds to study the effects of cyclic dynamic compressive loading on chondrocyte gene expression and extracellular matrix (ECM production. Dynamic culture of porcine chondrocytes was studied at 1 Hz, 10% to 40% strain and 1 to 9 h/day stimulation duration, in a mechanical-driven multi-chamber bioreactor for 14 days. From the experimental results, we could identify the optimum dynamic culture condition (20% and 3 h/day to enhance the chondrocytic phenotype of chondrocytes from the expression of marker (Col I, Col II, Col X, TNF-α, TGF-β1 and IGF-1 genes by quantitative real-time polymerase chain reactions (qRT-PCR and production of ECM (GAGs and Col II by biochemical analysis and immunofluorescence staining. With up-regulated growth factor (TGF-β1 and IGF-1 genes, co-culture of chondrocytes with porcine adipose-derived stem cells (ASCs was employed to facilitate chondrogenic differentiation of ASCs during dynamic culture in cryogel scaffolds. By replacing half of the chondrocytes with ASCs during co-culture, we could obtain similar production of ECM (GAGs and Col II and expression of Col II, but reduced expression of Col I, Col X and TNF-α. Subcutaneous implantation of cells/scaffold constructs in nude mice after mono-culture (chondrocytes or ASCs or co-culture (chondrocytes + ASCs and subject to static or dynamic culture condition in vitro for 14 days was tested for tissue-engineering applications. The constructs were retrieved 8 weeks post-implantation for histological analysis by Alcian blue, Safranin O and Col II immunohistochemical staining. The most abundant ectopic cartilage tissue was found for the chondrocytes and chondrocytes + ASCs groups using dynamic culture, which showed similar neo-cartilage formation capability with half of the

Study on postpartum estrus of guinea pigs (Cavia cobaya using Anredera cordifolia leaf extract

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D. Wijayanti

2017-04-01

Full Text Available Aim: The purpose of this study was to determine the postpartum estrus cycle of guinea pigs (Cavia cobaya using Anredera cordifolia leaf extract. Materials and Methods: Materials used were 8 males and 8 females of C. cobaya with body weight ranged 400-450 g. Mating ratio applied was 1:1. Treatments given were 0, 10, 50 and 90 mg of A. cordifolia leaf extract/head, designated as T0, T1, T2 and T3, respectively. Two females were subjected for each treatment. A. cordifolia leaf extract was administered orally from 10 days prepartum to 10 days postpartum. Observation of mating behavior, vulva morphology, and vaginal smear preparation was done in the afternoon for 10 days subsequence postpartum. Data were analyzed by univariate method and descriptively. Results: The results showed that the addition of A. cordifolia leaf extract 50 mg orally could accelerate the time of postpartum estrus based on the average frequency of mating behavior consisting of behavioral approach, allow the buck to sniffing her, mating positions, standing heat, lordosis position, and copulation. During estrus, vulva morphology was red color, had much mucus and no thin membrane covering vagina. There were a lot of superficial cells on vagina. Conclusion: The best treatment to accelerate occurring postpartum estrus was the addition of A. cordifolia leaf extract as many as 50 mg/head weight (T2 orally.

Isolation and determination of cultural characteristics of a new highly CO2 tolerant fresh water microalgae

International Nuclear Information System (INIS)

Yue Lihong; Chen Weigong

2005-01-01

Fresh water microalgae, which has high CO 2 tolerance, were isolated and its cultural characteristics were investigated. The ZY-1 strain was identified as genus Chlorella. It showed maximum growth at 10% (v/v) CO 2 enriched air flowing condition, and a good growth rate in a broad range of physically controllable conditions, including CO 2 concentration up to 70% (v/v), CO 2 enriched air flow rate, temperature and pH value. The results indicated the feasibility of the ZY-1 strain for fixing CO 2 from stack gases

Vibrios and Aeromonas.

Science.gov (United States)

Holmberg, S D

1988-09-01

There are many similarities in the Vibrionaceae that cause human illness in the United States (see Table 1). Vibrios are characteristically indigenous to marine, estuarine, and brackish environments. They are distributed mainly in Gulf of Mexico coastal water, and these organisms "bloom" when the water is warm. Outbreaks of disease in humans frequently occur in summer, coinciding with multiplication of vibrios in warm water. Sporadic cases and small outbreaks of cholera continue to occur in persons living on or near the Gulf of Mexico, but infection in most persons is unrecognized. In fact, more serious and frequent illnesses result from V. vulnificus wound infections and from gastroenteritis caused by vibrios other than V. cholerae 01. Underlying hepatic or neoplastic disease (especially leukemia) apparently increases the likelihood and severity of illnesses caused by V. vulnificus and Aeromonas. Some Vibrionaceae produce clinical illness by means of enterotoxins identical or similar to cholera toxin. For many others, hemolysins, cytotoxins, and other exotoxins are necessary to produce disease; the importance of these virulence factors often is not known or the importance of these virulence factors often is not known or is of doubtful significance. Also, purported pathogenicity as demonstrated by animal models, such as fluid accumulation in ligated ileal loops, is quite nonspecific and needs to be interpreted cautiously. For Plesiomonas, a mode of pathogenesis has not been discovered. Eating raw shellfish (frequently raw oysters) has been linked epidemiologically to enteric infections with most of these bacteria; foreign travel and exposure to seawater are other frequently observed epidemiologic associations with infection. Foreign travel, particularly to the Yucatan Peninsula of Mexico, has been strongly associated with the acquisition of non-01 V. cholerae and Plesiomonas organisms. Most Vibrionaceae in the United States are susceptible in vitro--and illnesses

Metabolomics Investigation of an Association of Induced Features and Corresponding Fungus during the Co-culture of Trametes versicolor and Ganoderma applanatum

Science.gov (United States)

Xu, Xiao-Yan; Shen, Xiao-Ting; Yuan, Xiao-Jie; Zhou, Yuan-Ming; Fan, Huan; Zhu, Li-Ping; Du, Feng-Yu; Sadilek, Martin; Yang, Jie; Qiao, Bin; Yang, Song

2018-01-01

The co-culture of Trametes versicolor and Ganoderma applanatum is a model of intense basidiomycete interaction, which induces many newly synthesized or highly produced features. Currently, one of the major challenges is an identification of the origin of induced features during the co-culture. Herein, we report a 13C-dynamic labeling analysis used to determine an association of induced features and corresponding fungus even if the identities of metabolites were not available or almost nothing was known of biochemical aspects. After the co-culture of T. versicolor and G. applanatum for 10 days, the mycelium pellets of T. versicolor and G. applanatum were sterilely harvested and then mono-cultured in the liquid medium containing half fresh medium with 13C-labeled glucose as carbon source and half co-cultured supernatants collected on day 10. 13C-labeled metabolome analyzed by LC-MS revealed that 31 induced features including 3-phenyllactic acid and orsellinic acid were isotopically labeled in the mono-culture after the co-culture stimulation. Twenty features were derived from T. versicolor, 6 from G. applanatum, and 5 features were synthesized by both T. versicolor and G. applanatum. 13C-labeling further suggested that 12 features such as previously identified novel xyloside [N-(4-methoxyphenyl)formamide 2-O-beta-D-xyloside] were likely induced through the direct physical interaction of mycelia. Use of molecular network analysis combined with 13C-labeling provided an insight into the link between the generation of structural analogs and producing fungus. Compound 1 with m/z 309.0757, increased 15.4-fold in the co-culture and observed 13C incorporation in the mono-culture of both T. versicolor and G. applanatum, was purified and identified as a phenyl polyketide, 2,5,6-trihydroxy-4, 6-diphenylcyclohex-4-ene-1,3-dione. The biological activity study indicated that this compound has a potential to inhibit cell viability of leukemic cell line U937. The current work sets

Antimicrobial activity of some of the south-Indian spices against serotypes of Escherichia coli, Salmonella, Listeria monocytogenes and Aeromonas hydrophila Atividade antimicrobiana de condimentos do sul da India sobre Escherichia coli, Salmonella, Listeria monocytogenes e Aeromonas hydrophila

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M.N. Indu

2006-06-01

Full Text Available Antibacterial activity of extracts of Allium sativum (garlic, Myristica fragrans (nutmeg, Zingiber officinale (ginger, Allium cepa (onion and Piper nigrum (pepper has been evaluated against 20 different serogroups of Escherichia coli, 8 serotypes of Salmonella, Listeria monocytogenes and Aeromonas hydrophila. Garlic extract showed excellent antibacterial activity against all the test organisms, except L. monocytogenes. Nutmeg showed good anti-listerial activity, although activity against E. coli and Salmonella were serotype dependent. Both garlic and nutmeg extracts were effective against A. hydrophila. Extracts of ginger showed inhibitory activity against two serogroups of E. coli: as O8 (enterotoxigenic E. coli and O88 only. Extracts of onion and pepper did not show any antibacterial activity against the test organisms.Avaliou-se a atividade antimicrobiana de extratos de alho (Allium sativum, noz-moscada (Mysritica frangrans, gengibre (Zingiber officinale cebola (Allium cepa e pimenta do reino (Piper nigrum sobre 20 sorotipos de Escherichia coli, 8 sorotipos de Salmonella, Listeria monocytogenes e Aeromonas hydrophila. O alho apresentou atividade antimicrobiana excelente sobre todos os microrganismos testados, excepto L. monocytogenes. A noz-moscada apresentou boa atividade antilisteria, emboara atividade sobre E. coli e Salmonella tenha sido sorotipo-dependente. Tanto alho como noz-moscada foram eficientes contra A. hydrophila. O extrato de gengibre apresentou atividade inibitória sobre dois sorotipos de E. coli: 08 (enterotoxigenico e 088. Os extratos de cebola e pimenta do reino não apresentaram nenhuma atividade contra os microrganismos testados.

Temperature dependence of bioelectrochemical CO2 conversion and methane production with a mixed-culture biocathode.

Science.gov (United States)

Yang, Hou-Yun; Bao, Bai-Ling; Liu, Jing; Qin, Yuan; Wang, Yi-Ran; Su, Kui-Zu; Han, Jun-Cheng; Mu, Yang

2018-02-01

This study evaluated the effect of temperature on methane production by CO 2 reduction during microbial electrosynthesis (MES) with a mixed-culture biocathode. Reactor performance, in terms of the amount and rate of methane production, current density, and coulombic efficiency, was compared at different temperatures. The microbial properties of the biocathode at each temperature were also analyzed by 16S rRNA gene sequencing. The results showed that the optimum temperature for methane production from CO 2 reduction in MES with a mixed-culture cathode was 50°C, with the highest amount and rate of methane production of 2.06±0.13mmol and 0.094±0.01mmolh -1 , respectively. In the mixed-culture biocathode MES, the coulombic efficiency of methane formation was within a range of 19.15±2.31% to 73.94±2.18% due to by-product formation at the cathode, including volatile fatty acids and hydrogen. Microbial analysis demonstrated that temperature had an impact on the diversity of microbial communities in the biofilm that formed on the MES cathode. Specifically, the hydrogenotrophic methanogen Methanobacterium became the predominant archaea for methane production from CO 2 reduction, while the abundance of the aceticlastic methanogen Methanosaeta decreased with increased temperature. Copyright © 2017. Published by Elsevier B.V.

Morphological analysis of human umbilical vein endothelial cells co-cultured with ovarian cancer cells in 3D: An oncogenic angiogenesis assay.

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Xiao Wan

Full Text Available Antiangiogenic therapy for cancer is a strategy targeted at tumour vasculature, often in combination with conventional cytotoxicity treatments. Animal testing is still the most common method used for evaluating the efficacy of new drugs but tissue-engineered in vitro models are becoming more acceptable for replacing and reducing the use of animals in anti-cancer drug screening. In this study, a 3D co-culture model of human endothelial cells and ovarian cancer cells was developed. This model has the potential to mimic the interactions between endothelial cells and ovarian cancer cells. The feasibility of applying this model in drug testing was explored here. The complex morphology of the co-culture system, which features development of both endothelial tubule-like structures and tumour structures, was analysed quantitatively by an image analysis method. The co-culture morphology integrity was maintained for 10 days and the potential of the model for anti-cancer drug testing was evaluated using Paclitaxel and Cisplatin, two common anti-tumour drugs with different mechanisms of action. Both traditional cell viability assays and quantitative morphological analyses were applied in the drug testing. Cisplatin proved a good example showing the advantages of morphological analysis of the co-culture model when compared with mono-culture of endothelial cells, which did not reveal an inhibitory effect of Cisplatin on the tubule-like endothelial structures. Thus, the tubule areas of the co-culture reflected the anti-angiogenesis potential of Cisplatin. In summary, in vitro cancer models can be developed using a tissue engineering approach to more closely mimic the characteristics of tumours in vivo. Combined with the image analysis technique, this developed 3D co-culture angiogenesis model will provide more reproducible and reliably quantified results and reveal further information of the drug's effects on both tumour cell growth and tumour angiogenesis.

ANTIBIOTICS RESISTANCE AND PUTATIVE VIRULENCE FACTORS OF AEROMONAS HYDROPHILA ISOLATED FROM ESTUARY

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Olumide Adedokun Odeyemi

2012-06-01

Full Text Available This study aim to investigate antibiotics resistance profile and putative virulence factors of Aeromonas hydrophila isolated from estuary. Bacteria used for this study were isolated from water and sediment samples obtained from Sungai Melayu, Johor, Malaysia. Serially diluted 100 µL water and 1g sediment were inoculated on modified Rimler - Shott (mRS agar. Colonies with distinct cultural characteristics were picked for further studies. Isolates were tested for biofilm productions, protease enzyme and antibiotics resistance profile using agar well diffusion method against 10 commercial antibiotics. Congo Red Agar (CRA, Microplate and Standard Tube (ST methods were used for assessment of biofilm formation among the isolates while Skim Milk Agar was used for protease production. Sw.KMJ 3 and Sw.KMJ 9 produced black crystalline colonies on CRA. Six of the isolates were biofilm producers in ST method. Result of Microplate method, helped in grouping the isolates into weak (n = 8, moderate (n = 3 and strong producers (n = 4 at 540 nm wavelength. All the isolates were classified as weak ODc  ODi 0.1, moderate ODi = 0.1  0.12 and strong producers ODi  0.12 respectively at 540 nm wavelength. Antibiotics susceptibility test also revealed that all the isolates were resistant to between 6 and 10 antibiotics. Two isolates each were resistant to 6 (60 %, 7 (70 % and 9 (90 % antibiotics respectively. Eight of the isolates showed resistance to 8 (80 % antibiotics while only isolate Sw.KMJ-7 showed resistance to all the tested antibiotics. Sw.KMJ-3, Sw.KMJ-8 and Sw.KMJ-9 produced protease enzyme on SMA. The isolates were also found to be resistant to both antibiotics and heavy metals.

Mixed Wastewater Coupled with CO2 for Microalgae Culturing and Nutrient Removal.

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Lili Yao

Full Text Available Biomass, nutrient removal capacity, lipid productivity and morphological changes of Chlorella sorokiniana and Desmodesmus communis were investigated in mixed wastewaters with different CO2 concentrations. Under optimal condition, which was 1:3 ratio of swine wastewater to second treated municipal wastewater with 5% CO2, the maximum biomass concentrations were 1.22 g L-1 and 0.84 g L-1 for C. sorokiniana and D. communis, respectively. Almost all of the ammonia and phosphorus were removed, the removal rates of total nitrogen were 88.05% for C. sorokiniana and 83.18% for D. communis. Lipid content reached 17.04% for C. sorokiniana and 20.37% for D. communis after 10 days culture. CO2 aeration increased intracellular particle numbers of both microalgae and made D. communis tend to be solitary. The research suggested the aeration of CO2 improve the tolerance of microalgae to high concentration of NH4-N, and nutrient excess stress could induce lipid accumulation of microalgae.

Aeromonas salmonicida subsp. salmonicida strains isolated from Chinese freshwater fish contain a novel genomic island and possible regional-specific mobile genetic elements profiles

DEFF Research Database (Denmark)

Long, Meng; Nielsen, Tue K; Leisner, Jørgen J

2016-01-01

Two strains of Aeromonas salmonicida, YK and BG, were isolated from largemouth bronze gudgeon and northern whitefish in China, and identified as A. salmonicida subsp. salmonicida based on phylogenetic analysis of vapA and 16S rRNA gene sequences. YK and BG originated from freshwater fish, one...

A Triple Co-Culture Model of the Human Respiratory Tract to Study Immune-Modulatory Effects of Liposomes and Virosomes.

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Rebecca A M Blom

Full Text Available The respiratory tract with its ease of access, vast surface area and dense network of antigen-presenting cells (APCs represents an ideal target for immune-modulation. Bio-mimetic nanocarriers such as virosomes may provide immunomodulatory properties to treat diseases such as allergic asthma. In our study we employed a triple co-culture model of epithelial cells, macrophages and dendritic cells to simulate the human airway barrier. The epithelial cell line 16HBE was grown on inserts and supplemented with human blood monocyte-derived macrophages (MDMs and dendritic cells (MDDCs for exposure to influenza virosomes and liposomes. Additionally, primary human nasal epithelial cells (PHNEC and EpCAM+ epithelial progenitor cell mono-cultures were utilized to simulate epithelium from large and smaller airways, respectively. To assess particle uptake and phenotype change, cell cultures were analyzed by flow cytometry and pro-inflammatory cytokine concentrations were measured by ELISA. All cell types internalized virosomes more efficiently than liposomes in both mono- and co-cultures. APCs like MDMs and MDDCs showed the highest uptake capacity. Virosome and liposome treatment caused a moderate degree of activation in MDDCs from mono-cultures and induced an increased cytokine production in co-cultures. In epithelial cells, virosome uptake was increased compared to liposomes in both mono- and co-cultures with EpCAM+ epithelial progenitor cells showing highest uptake capacity. In conclusion, all cell types successfully internalized both nanocarriers with virosomes being taken up by a higher proportion of cells and at a higher rate inducing limited activation of MDDCs. Thus virosomes may represent ideal carrier antigen systems to modulate mucosal immune responses in the respiratory tract without causing excessive inflammatory changes.

Expression of HSP27, HSP72 and MRP proteins in in vitro co-culture ...

Indian Academy of Sciences (India)

Prakash

2009-12-09

Dec 9, 2009 ... in co-cultures of human colon carcinoma cell spheroids obtained from different grades of ...... of adherens junction in prostate epithelial cells; Exp. Cell. Res. ... tumour-associated antigens is related to estrogen receptor status;.

Effectiveness of ambon banana stem juice as immunostimulatory against Aeromonas hydrophila infections in catfish Clarias gariepinus

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Qorie Astria

2017-07-01

Full Text Available ABSTRACT Outbreaks of infectious diseases due to Aeromonas hydrophila in catfish can cause high death rates (80–100%. Fish disease control can be done using phytopharmaceutical to prevent or treat diseases of fish. One of the phytopharmaceutical that known to prevent the fish diseases is ambon banana stem Musa cavendishii var. dwarf Paxton. This study was conducted to test the effectiveness of catfish immersion using banana stem juice as an immunostimulant against bacterial infections A. hydrophila. The observed parameters were a total performance of production, hematological test, and water quality. Immersion of catfish seeds with stem juice was performed at a concentration of 5 mL/L, 13 mL/L, and 21 mL/L for 30 minutes. Each treatment consisted of three replications. A  total of 15 fishes were immersed in 1.5 L water. At day-9 after immersion, fish was infected by A. hydrophila bacteria at a dose of 104 cfu/mL. The results showed that fish treated with banana stem juice at a concentration of 13 mL/L had the survival rate of 53.33±6.67% which is higher than positive control (33.33±6.67%. Based on hematology observations on the 3rd day post-challenge test (H12 with A. hydrophila, total erythrocytes, hemoglobin, total leukocytes, phagocytic activity, differential leukocyte, and lysozyme activity was highest in treatment13 mL/L. Lower feed conversion ratio (1.08±0.04 were also obtained in treatment 13 mL/L. Thus Immersion of seeds in stem juice can boost the immune system against infections A. hydrophila. Keywords: Aeromonas hydrophila, ambon banana stem juice, soaking, catfish, immune system  ABSTRAK Wabah penyakit akibat infeksi Aeromonas hydrophila pada ikan lele dapat menyebabkan tingkat kematian yang tinggi (80–100%. Penanggulangan penyakit ikan dapat dilakukan menggunakan fitofarmaka untuk mencegah ataupun mengobati penyakit ikan. Salah satu fitofarmaka yang dapat digunakan dalam upaya pencegahan penyakit ikan adalah batang

Chitosan-hyaluronan based 3D co-culture platform for studying the crosstalk of lung cancer cells and mesenchymal stem cells.

Science.gov (United States)

Han, Hao-Wei; Hsu, Shan-Hui

2016-09-15

The controversial roles of mesenchymal stem cells (MSCs) in lung cancer development are not yet resolved because of the lack of an extracellular environment that mimics the tumor microenvironment. Three-dimensional (3D) culture system is an emerging research tool for biomedical applications such as drug screening. In this study, MSCs and human non-small cell lung carcinoma cells (A549) were co-cultured on a thin biomaterial-based substratum (hyaluronan-grafted chitosan, CS-HA; ∼2μm), and they were self-organized into the 3D tumor co-spheroids with core-shell structure. The gene expression levels of tumorigenicity markers in cancer cells associated with cancer stemness, epithelial-mesenchymal transition (EMT) property, and cell mobility were up-regulated for more than twofold in the MSC-tumor co-spheroids, through the promoted expression of certain tumor enhancers and the direct cell-cell interaction. To verify the different extents of tumorigenicity, A549 cells or those co-cultured with MSCs were transplanted into zebrafish embryos for evaluation in vivo. The tumorigenicity obtained from the zebrafish xenotransplantation model was consistent with that observed in vitro. These evidences suggest that the CS-HA substrate-based 3D co-culture platform for cancer cells and MSCs may be a convenient tool for studying the cell-cell interaction in a tumor-like microenvironment and potentially for cancer drug testing. Mesenchymal stem cells (MSCs) have been found in several types of tumor tissues. However, the controversial roles of MSCs in cancer development are still unsolved. Chitosan and hyaluronan are commonly used materials in the biomedical field. In the current study, we co-cultured lung cancer cells and MSCs on the planar hyaluronan-grafted chitosan (CS-HA) hybrid substrates, and discovered that lung cancer cells and MSCs were rapidly self-assembled into 3D tumor spheroids with core-shell structure on the substrates after only two days in culture. Therefore, CS

Rethinking Sponge Bob and Ninja Turtles: Popular Culture as Funds of Knowledge for Curriculum Co-Construction

Science.gov (United States)

Hedges, Helen

2011-01-01

Children's interest in popular culture was clear in my study of interests-based curriculum. Yet, perhaps unsurprisingly, it was a contentious site of curriculum co-construction. This article explores this tension. It argues that interpreting popular culture as "funds of knowledge" might assist teachers to consider a different view of this interest…

Cultural, social and intrapersonal factors associated with clusters of co-occurring health-related behaviours among adolescents.

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Klein Velderman, Mariska; Dusseldorp, Elise; van Nieuwenhuijzen, Maroesjka; Junger, Marianne; Paulussen, Theo G W M; Reijneveld, Sijmen A

2015-02-01

Adverse health-related behaviours (HRBs) have been shown to co-occur in adolescents. Evidence lacks on factors associated with these co-occurring HRBs. The Theory of Triadic Influence (TTI) offers a route to categorize these determinants according to type (social, cultural and intrapersonal) and distance in the causal pathway (ultimate or distal). Our aims were to identify cultural, social and intrapersonal factors associated with co-occurring HRBs and to assess the relative importance of ultimate and distal factors for each cluster of co-occurring HRBs. Respondents concerned a random sample of 898 adolescents aged 12-18 years, stratified by age, sex and educational level of head of household. Data were collected via face-to-face computer-assisted interviewing and internet questionnaires. Analyses were performed for young (12-15 years) and late (16-18 years) adolescents regarding two and three clusters of HRB, respectively. For each cluster of HRBs (e.g. smoking, delinquency), associated factors were found. These accounted for 27 to 57% of the total variance per cluster. Factors came in particular from the intrapersonal stream of the TTI at the ultimate level and the social stream at the distal level. Associations were strongest for parenting practices, risk behaviours of friends and parents and self-control. Results of this study confirm that it is possible to identify a selection of cultural, social and intrapersonal factors associated with co-occurring HRBs among adolescents. © The Author 2014. Published by Oxford University Press on behalf of the European Public Health Association. All rights reserved.

Procesos de construcción de la diferencia cultural en el co-manejo del Parque Nacional Nahuel Huapi

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Florencia Trentini

2016-01-01

Full Text Available This article has sought to present an ethnographic analysis of the processes of constructing cultural difference in the co-management of Nahuel Huapi National Park (Patagonia, Argentina. It focuses on the articulation of identity, culture and territory, understanding co-management to be a global-local conservationist arena in which multiple interests that reshape identities and environmental policies come into dispute. The study concludes that communities are not approached as historical products in this process, but rather as totalizing and homogeneous entities that must comply with predetermined attributes in order to be legitimized and incorporated into the co-management process. In contrast to this tendency, it shows how different “ways of being a community” require a degree of institutional plasticity that cannot be guaranteed with such rigid models of co-management.

CP-25 attenuates the inflammatory response of fibroblast-like synoviocytes co-cultured with BAFF-activated CD4(+) T cells.

Science.gov (United States)

Jia, Xiaoyi; Wei, Fang; Sun, Xiaojing; Chang, Yan; Xu, Shu; Yang, Xuezhi; Wang, Chun; Wei, Wei

2016-08-02

Total glucosides of paeony (TGP) is the first anti-inflammatory immune regulatory drug approved for the treatment of rheumatoid arthritis in China. A novel compound, paeoniflorin-6'-O-benzene sulfonate (code CP-25), comes from the structural modification of paeoniflorin (Pae), which is the effective active ingredient of TGP. The aim of the present study is to investigate the effect of CP-25 on adjuvant arthritis (AA) fibroblast-like synoviocytes (FLS) co-cultured with BAFF-activated CD4(+) T cells and the expression of BAFF-R in CD4(+) T cells. The mRNA expression of BAFF and its receptors was assessed by qPCR. The expression of BAFF receptors in CD4(+) T cells was analyzed by flow cytometry. The effect of CP-25 on AA rats was evaluated by their joint histopathology. The cell culture growth of thymocytes and FLS was detected by cell counting kit (CCK-8). The concentrations of IL-1β, TNF-α, and IL-6 were measured by Enzyme-linked immunosorbent assay (ELISA). The mRNA expression levels of BAFF and BAFF-R were enhanced in the mesenteric lymph nodes of AA rats, TACI expression was reduced, and BCMA had no change. The expression of BAFF-R in CD4(+) T cells was also enhanced. CP-25 alleviated the joint histopathology and decreased the expression of BAFF-R in CD4(+) T cells from AA rats in vivo. In vitro, CP-25 inhibited the abnormal cell culture growth of BAFF-stimulated thymocytes and FLS. In the co-culture system, IL-1β, IL-6 and TNF-α production was enhanced by FLS co-cultured with BAFF-activated CD4(+) T cells. Moreover, BAFF-stimulated CD4(+) T cells promoted the cell culture growth of FLS. The addition of CP-25 decreased the expression of BAFF-R in CD4(+) T cells and inhibited the cell culture growth and cytokine secretion ability of FLS co-cultured with BAFF-activated CD4(+) T cells. The present study indicates that CP-25 may repress the cell culture growth and cytokine secretion ability of FLS, and its inhibitory effects might be associated with its ability

Analysis of the Genome and Mobilome of a Dissimilatory Arsenate Reducing Aeromonas sp. O23A Reveals Multiple Mechanisms for Heavy Metal Resistance and Metabolism

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Witold Uhrynowski

2017-05-01

Full Text Available Aeromonas spp. are among the most ubiquitous microorganisms, as they have been isolated from different environmental niches including waters, soil, as well as wounds and digestive tracts of poikilothermic animals and humans. Although much attention has been paid to the pathogenicity of Aeromonads, the role of these bacteria in environmentally important processes, such as transformation of heavy metals, remains to be discovered. Therefore, the aim of this study was a detailed genomic characterization of Aeromonas sp. O23A, the first representative of this genus capable of dissimilatory arsenate reduction. The strain was isolated from microbial mats from the Zloty Stok mine (SW Poland, an environment strongly contaminated with arsenic. Previous physiological studies indicated that O23A may be involved in both mobilization and immobilization of this metalloid in the environment. To discover the molecular basis of the mechanisms behind the observed abilities, the genome of O23A (∼5.0 Mbp was sequenced and annotated, and genes for arsenic respiration, heavy metal resistance (hmr and other phenotypic traits, including siderophore production, were identified. The functionality of the indicated gene modules was assessed in a series of minimal inhibitory concentration analyses for various metals and metalloids, as well as mineral dissolution experiments. Interestingly, comparative analyses revealed that O23A is related to a fish pathogen Aeromonas salmonicida subsp. salmonicida A449 which, however, does not carry genes for arsenic respiration. This indicates that the dissimilatory arsenate reduction ability may have been lost during genome reduction in pathogenic strains, or acquired through horizontal gene transfer. Therefore, particular emphasis was placed upon the mobilome of O23A, consisting of four plasmids, a phage, and numerous transposable elements, which may play a role in the dissemination of hmr and arsenic metabolism genes in the

Analysis of the Genome and Mobilome of a Dissimilatory Arsenate Reducing Aeromonas sp. O23A Reveals Multiple Mechanisms for Heavy Metal Resistance and Metabolism.

Science.gov (United States)

Uhrynowski, Witold; Decewicz, Przemyslaw; Dziewit, Lukasz; Radlinska, Monika; Krawczyk, Pawel S; Lipinski, Leszek; Adamska, Dorota; Drewniak, Lukasz

2017-01-01

Aeromonas spp. are among the most ubiquitous microorganisms, as they have been isolated from different environmental niches including waters, soil, as well as wounds and digestive tracts of poikilothermic animals and humans. Although much attention has been paid to the pathogenicity of Aeromonads, the role of these bacteria in environmentally important processes, such as transformation of heavy metals, remains to be discovered. Therefore, the aim of this study was a detailed genomic characterization of Aeromonas sp. O23A, the first representative of this genus capable of dissimilatory arsenate reduction. The strain was isolated from microbial mats from the Zloty Stok mine (SW Poland), an environment strongly contaminated with arsenic. Previous physiological studies indicated that O23A may be involved in both mobilization and immobilization of this metalloid in the environment. To discover the molecular basis of the mechanisms behind the observed abilities, the genome of O23A (∼5.0 Mbp) was sequenced and annotated, and genes for arsenic respiration, heavy metal resistance ( hmr ) and other phenotypic traits, including siderophore production, were identified. The functionality of the indicated gene modules was assessed in a series of minimal inhibitory concentration analyses for various metals and metalloids, as well as mineral dissolution experiments. Interestingly, comparative analyses revealed that O23A is related to a fish pathogen Aeromonas salmonicida subsp. salmonicida A449 which, however, does not carry genes for arsenic respiration. This indicates that the dissimilatory arsenate reduction ability may have been lost during genome reduction in pathogenic strains, or acquired through horizontal gene transfer. Therefore, particular emphasis was placed upon the mobilome of O23A, consisting of four plasmids, a phage, and numerous transposable elements, which may play a role in the dissemination of hmr and arsenic metabolism genes in the environment. The obtained

Analysis of the Genome and Mobilome of a Dissimilatory Arsenate Reducing Aeromonas sp. O23A Reveals Multiple Mechanisms for Heavy Metal Resistance and Metabolism

Science.gov (United States)

Uhrynowski, Witold; Decewicz, Przemyslaw; Dziewit, Lukasz; Radlinska, Monika; Krawczyk, Pawel S.; Lipinski, Leszek; Adamska, Dorota; Drewniak, Lukasz

2017-01-01

Aeromonas spp. are among the most ubiquitous microorganisms, as they have been isolated from different environmental niches including waters, soil, as well as wounds and digestive tracts of poikilothermic animals and humans. Although much attention has been paid to the pathogenicity of Aeromonads, the role of these bacteria in environmentally important processes, such as transformation of heavy metals, remains to be discovered. Therefore, the aim of this study was a detailed genomic characterization of Aeromonas sp. O23A, the first representative of this genus capable of dissimilatory arsenate reduction. The strain was isolated from microbial mats from the Zloty Stok mine (SW Poland), an environment strongly contaminated with arsenic. Previous physiological studies indicated that O23A may be involved in both mobilization and immobilization of this metalloid in the environment. To discover the molecular basis of the mechanisms behind the observed abilities, the genome of O23A (∼5.0 Mbp) was sequenced and annotated, and genes for arsenic respiration, heavy metal resistance (hmr) and other phenotypic traits, including siderophore production, were identified. The functionality of the indicated gene modules was assessed in a series of minimal inhibitory concentration analyses for various metals and metalloids, as well as mineral dissolution experiments. Interestingly, comparative analyses revealed that O23A is related to a fish pathogen Aeromonas salmonicida subsp. salmonicida A449 which, however, does not carry genes for arsenic respiration. This indicates that the dissimilatory arsenate reduction ability may have been lost during genome reduction in pathogenic strains, or acquired through horizontal gene transfer. Therefore, particular emphasis was placed upon the mobilome of O23A, consisting of four plasmids, a phage, and numerous transposable elements, which may play a role in the dissemination of hmr and arsenic metabolism genes in the environment. The obtained

Co-culture of chondrocytes and bone marrow mesenchymal stem cells in vitro enhances the expression of cartilaginous extracellular matrix components

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Chang Qing

2011-04-01

Full Text Available Chondrocytes and bone marrow mesenchymal stem cells (BMSCs are frequently used as seed cells in cartilage tissue engineering. In the present study, we determined if the co-culture of rabbit articular chondrocytes and BMSCs in vitro promotes the expression of cartilaginous extracellular matrix and, if so, what is the optimal ratio of the two cell types. Cultures of rabbit articular chondrocytes and BMSCs were expanded in vitro and then cultured individually or at a chondrocyte:BMSC ratio of 4:1, 2:1, 1:1, 1:2, 1:4 for 21 days and cultured in DMEM/F12. BMSCs were cultured in chondrogenic induction medium. Quantitative real-time RT-PCR and Western blot were used to evaluate gene expression. In the co-cultures, type II collagen and aggrecan expression increased on days 14 and 21. At the mRNA level, the expression of type II collagen and aggrecan on day 21 was much higher in the 4:1, 2:1, and 1:1 groups than in either the articular chondrocyte group or the induced BMSC group, and the best ratio of co-culture groups seems to be 2:1. Also on day 21, the expression of type II collagen and aggrecan proteins in the 2:1 group was much higher than in all other groups. The results demonstrate that the co-culture of rabbit chondrocytes and rabbit BMSCs at defined ratios can promote the expression of cartilaginous extracellular matrix. The optimal cell ratio appears to be 2:1 (chondrocytes:BMSCs. This approach has potential applications in cartilage tissue engineering since it provides a protocol for maintaining and promoting seed-cell differentiation and function.

Co-culture of chondrocytes and bone marrow mesenchymal stem cells in vitro enhances the expression of cartilaginous extracellular matrix components.

Science.gov (United States)

Qing, Chang; Wei-ding, Cui; Wei-min, Fan

2011-04-01

Chondrocytes and bone marrow mesenchymal stem cells (BMSCs) are frequently used as seed cells in cartilage tissue engineering. In the present study, we determined if the co-culture of rabbit articular chondrocytes and BMSCs in vitro promotes the expression of cartilaginous extracellular matrix and, if so, what is the optimal ratio of the two cell types. Cultures of rabbit articular chondrocytes and BMSCs were expanded in vitro and then cultured individually or at a chondrocyte:BMSC ratio of 4:1, 2:1, 1:1, 1:2, 1:4 for 21 days and cultured in DMEM/F12. BMSCs were cultured in chondrogenic induction medium. Quantitative real-time RT-PCR and Western blot were used to evaluate gene expression. In the co-cultures, type II collagen and aggrecan expression increased on days 14 and 21. At the mRNA level, the expression of type II collagen and aggrecan on day 21 was much higher in the 4:1, 2:1, and 1:1 groups than in either the articular chondrocyte group or the induced BMSC group, and the best ratio of co-culture groups seems to be 2:1. Also on day 21, the expression of type II collagen and aggrecan proteins in the 2:1 group was much higher than in all other groups. The results demonstrate that the co-culture of rabbit chondrocytes and rabbit BMSCs at defined ratios can promote the expression of cartilaginous extracellular matrix. The optimal cell ratio appears to be 2:1 (chondrocytes:BMSCs). This approach has potential applications in cartilage tissue engineering since it provides a protocol for maintaining and promoting seed-cell differentiation and function.

Effect of diisopropylfluorophosphate on synaptic transmission and acetylcholine sensitivity in neuroblastoma-myotube co-culture

International Nuclear Information System (INIS)

Adler, M.; Chang, F.C.T.; Foster, R.E.; Glenn, J.F.; Mark, G.; Maxwell, D.

1986-01-01

The authors investigate the effects of the irreversible organophosphorous cholinesterase inhibitor, DFP, on clonal G8-1 myotubes co-cultured with ACh- secreting NG108-15 neuroblastoma x glioma hybrid cells. The enzyme activity is shown, plotted as a function of time in culture. The enzyme activity remained low over four days. At the end of this time, the cultures were nearly confluent with myoblasts but contained less than 2% multinucleated myotubes. The AChE activity increased gradually after horse serum was added to the growth medium to promote myotube formation, reaching a maximum of 1.1 nmole. C 14 ACh/min/mg protein on the 15th day

Therapeutic effects of marshmallow (Althaea officinalis L.) extract on plasma biochemical parameters of common carp infected with Aeromonas hydrophila

OpenAIRE

Banaee, Mahdi; Soleimany, Vahid; Nematdoost Haghi, Behzad

2017-01-01

This study evaluated preclinical and clinical safety of marshmallow (Althaea officinalis L.) extract as a naturopathic medicine in common carp deliberately infected with Aeromonas hydrophila. The fish were fed 0 (control), 2.50, 5.00 and 10.00 g of marshmallow extract for 60 days in a preclinical experiment and then, challenged with A. hydrophila for a 10-day experiment. Significant increases were observed in aspartate aminotransferase (AST), lactate dehydrogenase (LDH), alkaline phosphatase ...

VAKSINASI IKAN TILAPIA (Oreochromis niloticus MENGGUNAKAN VAKSIN MONOVALEN DAN BIVALEN UNTUK PENCEGAHAN PENYAKIT MOTILE AEROMONAS SEPTICEMIA DAN STREPTOCOCCOSIS

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Desy Sugiani

2013-08-01

Full Text Available Peningkatan respon antibodi pascavaksinasi dengan antigen tunggal dan campuran dari bakterin Aeromonas hydrophila and Streptococcus agalactiae diharapkan dapat meningkatkan daya tahan ikan tilapia (Oreochromis niloticus terhadap penyakit Motile Aeromonas Septicemia/MAS dan Streptococcosis. Sediaan vaksin disiapkan dengan metode pembuatan dan formula yang berbeda, proses inaktifasi dilakukan dengan menambahkan 3% Neutral Buffer Formalin (NBF 10% pada biakan bakteri dalam media tumbuh BHI dan TSB. Vaksinasi diberikan melalui injeksi intraperitoneal dengan sediaan vaksin monovalen A. hydrophila, monovalen S. agalactiae, dan bivalen A. hydrophila + S. agalactiae (Sel utuh, produk ektraselular/ECP, crude supernatan, campuran sel utuh + ECP, dan broth. Uji tantang dilakukan menggunakan dosis LD50 infeksi tunggal maupun ko-infeksi dari bakteri A. hydrophila dan S. agalactiae. Efektivitas dan keampuhan vaksin tersebut dihitung berdasarkan nilai RPS (Relative Percent Survival dan hasil respon hematologi. Titer antibodi dapat terdeteksi setelah satu minggu pemeliharaan pasca vaksinasi. Nilai titer antar perlakuan vaksin bivalen berbeda nyata (P<0.05 dengan vaksin monovalen dan kontrol. Nilai RPS vaksin bivalen (campuran sel utuh + ECP mencapai 100 untuk uji tantang dengan A. hydrophila dan 56,7 pada uji tantang ko-infeksi. Vaksin monovalen A. hydrophila maupun S. agalactiae hanya mampu memproteksi terhadap bakteri homolog, tidak terjadi proteksi silang di antara keduanya.

UJI AKTIVITAS ANTIBAKTERI EKSTRAK RUMPUT LAUT Kappaphycus alvarezii DAN Eucheuma denticullatum TERHADAP BAKTERI Aeromonas hydrophila DAN Vibrio harveyii

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Dwi Budi Wiyanto

2010-04-01

Full Text Available Penelitian tentang Uji Aktivitas Antibakteri Ekstrak Rumput Laut Kappaphycus alvarezii dan Eucheuma denticullatum Terhadap Bakteri Aeromonas hydrophila dan Vibrio harveyii dilakukan, mengingat banyak dijumpai  penyakit pada usaha budidaya ikan dan udang, terutama bakteri Aeromonas hydrophila dan Vibrio harveyii. Penelitian ini bertujuan untuk mengetahui efektifitas senyawa bioaktif rumput laut K. alvarezii dan E. denticullatum yang diekstrak menggunakan pelarut metanol dan etanol sebagai antibakteri terhadap A. hydrophila dan V. harveyii.  Penelitian dilakukan sebanyak dua tahap, yaitu: (1 Uji aktivitas antibakteri ekstrak rumput laut; dan (2 Analisa senyawa bioaktif yang terdapat pada ekstrak rumput laut, dimana masing-masing tahapan dianalisis menggunakan rancangan acak lengkap faktorial. Hasil penelitian menunjukkan, dua jenis ekstrak rumput laut dengan pelarut metanol dan etanol, mempunyai daya antibakteri terhadap A. hydrophila dan V. harveyii. Ekstrak E. denticullatum dengan pelarut metanol memiliki daya hambat lebih luas dibanding ekstrak K. alvarezii dengan pelarut metanol terhadap A. hydrophila (19.43±0,55 mm. Ekstrak E. denticullatum dengan pelarut metanol memiliki daya hambat lebih luas dibanding ekastrak K. alvarezii dengan pelarut metanol terhadap V. harveyii (19.85±0,23 mm. Asam heksadekanoat merupakan senyawa paling dominan dijumpai pada ekstrak rumput laut K. alvarezii, dan E. denticullatum yang diekstrak menggunakan pelarut metanol. Kata Kunci : Aktivitas Antibakteri, Rumput laut K. alvarezii dan E. denticullatum,  Bakteri A.  hydrophila dan V. harveyii. 

Primary Neuron/Astrocyte Co-Culture on Polyelectrolyte Multilayer Films: A Template for Studying Astrocyte-Mediated Oxidative Stress in Neurons**

OpenAIRE

Kidambi, Srivatsan; Lee, Ilsoon; Chan, Christina

2008-01-01

We engineered patterned co-cultures of primary neurons and astrocytes on polyelectrolyte multilayer (PEM) films without the aid of adhesive proteins/ligands to study the oxidative stress mediated by astrocytes on neuronal cells. A number of studies have explored engineering co-culture of neurons and astrocytes predominantly using cell lines rather than primary cells owing to the difficulties involved in attaching primary cells onto synthetic surfaces. To our knowledge this is the first demons...

A First Generation Comparative Chromosome Map between Guinea Pig (Cavia porcellus) and Humans.

Science.gov (United States)

Romanenko, Svetlana A; Perelman, Polina L; Trifonov, Vladimir A; Serdyukova, Natalia A; Li, Tangliang; Fu, Beiyuan; O'Brien, Patricia C M; Ng, Bee L; Nie, Wenhui; Liehr, Thomas; Stanyon, Roscoe; Graphodatsky, Alexander S; Yang, Fengtang

2015-01-01

The domesticated guinea pig, Cavia porcellus (Hystricomorpha, Rodentia), is an important laboratory species and a model for a number of human diseases. Nevertheless, genomic tools for this species are lacking; even its karyotype is poorly characterized. The guinea pig belongs to Hystricomorpha, a widespread and important group of rodents; so far the chromosomes of guinea pigs have not been compared with that of other hystricomorph species or with any other mammals. We generated full sets of chromosome-specific painting probes for the guinea pig by flow sorting and microdissection, and for the first time, mapped the chromosomal homologies between guinea pig and human by reciprocal chromosome painting. Our data demonstrate that the guinea pig karyotype has undergone extensive rearrangements: 78 synteny-conserved human autosomal segments were delimited in the guinea pig genome. The high rate of genome evolution in the guinea pig may explain why the HSA7/16 and HSA16/19 associations presumed ancestral for eutherians and the three syntenic associations (HSA1/10, 3/19, and 9/11) considered ancestral for rodents were not found in C. porcellus. The comparative chromosome map presented here is a starting point for further development of physical and genetic maps of the guinea pig as well as an aid for genome assembly assignment to specific chromosomes. Furthermore, the comparative mapping will allow a transfer of gene map data from other species. The probes developed here provide a genomic toolkit, which will make the guinea pig a key species to unravel the evolutionary biology of the Hystricomorph rodents.

The effect of 60Co γ irradiation on culture in vitro of Pogostemon cablin (blanco) benth

International Nuclear Information System (INIS)

Wu Lirong; He Hong; Zhang Yanling; Liu Xing; Lin Xiaohua

2008-01-01

The 60 Co γ-irradiation on culture in vitro of Pogostemon cablin is studied to breed new variety. The explants used for 60 Co γ-irradiation were the leaf segments, nodular stem segments, stem segments and root tips from Pogostemon cablin. The media for culture in vitro were MT basal media containing 0.05mg/L BA. As irradiation dosage increased, the mortality of different explants from Pogostemon cablin increased. The irradiation dosage of LD50 for leaf segments, nodular stem segments and stem segments were 72, 64 and 66Gy, respectively, which were deduced from the regression equation. As the irradiation dosage increased, the ability of the plant regeneration of the explants decreased. 60 Co γ-irradiation could make the explants damage or die. Irradiation could inhibit the regeneration of explants from Pogostemon cablin obviously. The changes in the appearance of the regenerated plants were also observed. (authors)

Yeast-yeast interactions revealed by aromatic profile analysis of Sauvignon Blanc wine fermented by single or co-culture of non-Saccharomyces and Saccharomyces yeasts.

Science.gov (United States)

Sadoudi, Mohand; Tourdot-Maréchal, Raphaëlle; Rousseaux, Sandrine; Steyer, Damien; Gallardo-Chacón, Joan-Josep; Ballester, Jordi; Vichi, Stefania; Guérin-Schneider, Rémi; Caixach, Josep; Alexandre, Hervé

2012-12-01

There has been increasing interest in the use of selected non-Saccharomyces yeasts in co-culture with Saccharomyces cerevisiae. The main reason is that the multistarter fermentation process is thought to simulate indigenous fermentation, thus increasing wine aroma complexity while avoiding the risks linked to natural fermentation. However, multistarter fermentation is characterised by complex and largely unknown interactions between yeasts. Consequently the resulting wine quality is rather unpredictable. In order to better understand the interactions that take place between non-Saccharomyces and Saccharomyces yeasts during alcoholic fermentation, we analysed the volatile profiles of several mono-culture and co-cultures. Candida zemplinina, Torulaspora delbrueckii and Metschnikowia pulcherrima were used to conduct fermentations either in mono-culture or in co-culture with S. cerevisiae. Up to 48 volatile compounds belonging to different chemical families were quantified. For the first time, we show that C. zemplinina is a strong producer of terpenes and lactones. We demonstrate by means of multivariate analysis that different interactions exist between the co-cultures studied. We observed a synergistic effect on aromatic compound production when M. pulcherrima was in co-culture with S. cerevisiae. However a negative interaction was observed between C. zemplinina and S. cerevisiae, which resulted in a decrease in terpene and lactone content. These interactions are independent of biomass production. The aromatic profiles of T. delbrueckii and S. cerevisiae in mono-culture and in co-culture are very close, and are biomass-dependent, reflecting a neutral interaction. This study reveals that a whole family of compounds could be altered by such interactions. These results suggest that the entire metabolic pathway is affected by these interactions. Copyright © 2012 Elsevier Ltd. All rights reserved.

Phytochemical analysis of Binahong (Anredera Cordifolia) leaves extract to inhibit In Vitro growth of Aeromonas Hydrophila

Science.gov (United States)

Basyuni, Mohammad; Ginting, Prita Yulianti Anasta Br; Lesmana, Indra

2017-11-01

Binahong (Anredera cordifolia) is one of the medicinal plants commonly used to treat the disease of living organisms. The secondary metabolite of A. cordifolia leaves has been shown antibacterial activity. This study aimed to investigate the secondary metabolite of A. cordifolia leaves showing antibacterial and analysis the effectiveness of antibacterial to inhibit the growth of bacteria Aeromonas hydrophila. A paper disc soaked in a solution of A. cordifolia leaves extract was used to test in vitro at a concentration of 0% (w/v), 0.2%, 0.4%, 0.6%, 0.8%, and positive control of antibiotic (oxytetracycline), respectively. The extracts then placed on a tryptone soy agar (TSA) medium containing bacteria A. hydrophila and incubated at 37 °C for 24 hours. In vitro test showed that A. cordifolia leaves extract inhibited the growth of bacteria A. hydrophila with an inhibition area around the paper disc. The inhibition growth of A. hydrophila increased with the increasing of extract concentration. Bacterial growth was inhibited in the diameter zone of A. hydrophila under different levels of the extracts were 0 mm (0 % negative control), 8.4 mm (0.2 %), 9.4 mm (0.4 %), 10.5 mm (0.6 %), 11.9 mm (0.8 %), 27.5 mm (positive control), respectively. Phytochemical screening of A. cordifolia leaves extract indicated that the extracts contained flavonoid, phenol, saponin, alkaloid, triterpenoid, and β-sitosterol. Our in vitro study demonstrated the inhibition growth of A. hydrophila that caused the disease of motile Aeromonas septicemia (MAS).

Predictivity of dog co-culture model, primary human hepatocytes and HepG2 cells for the detection of hepatotoxic drugs in humans

International Nuclear Information System (INIS)

Atienzar, Franck A.; Novik, Eric I.; Gerets, Helga H.; Parekh, Amit; Delatour, Claude; Cardenas, Alvaro; MacDonald, James; Yarmush, Martin L.; Dhalluin, Stéphane

2014-01-01

Drug Induced Liver Injury (DILI) is a major cause of attrition during early and late stage drug development. Consequently, there is a need to develop better in vitro primary hepatocyte models from different species for predicting hepatotoxicity in both animals and humans early in drug development. Dog is often chosen as the non-rodent species for toxicology studies. Unfortunately, dog in vitro models allowing long term cultures are not available. The objective of the present manuscript is to describe the development of a co-culture dog model for predicting hepatotoxic drugs in humans and to compare the predictivity of the canine model along with primary human hepatocytes and HepG2 cells. After rigorous optimization, the dog co-culture model displayed metabolic capacities that were maintained up to 2 weeks which indicates that such model could be also used for long term metabolism studies. Most of the human hepatotoxic drugs were detected with a sensitivity of approximately 80% (n = 40) for the three cellular models. Nevertheless, the specificity was low approximately 40% for the HepG2 cells and hepatocytes compared to 72.7% for the canine model (n = 11). Furthermore, the dog co-culture model showed a higher superiority for the classification of 5 pairs of close structural analogs with different DILI concerns in comparison to both human cellular models. Finally, the reproducibility of the canine system was also satisfactory with a coefficient of correlation of 75.2% (n = 14). Overall, the present manuscript indicates that the dog co-culture model may represent a relevant tool to perform chronic hepatotoxicity and metabolism studies. - Highlights: • Importance of species differences in drug development. • Relevance of dog co-culture model for metabolism and toxicology studies. • Hepatotoxicity: higher predictivity of dog co-culture vs HepG2 and human hepatocytes

Predictivity of dog co-culture model, primary human hepatocytes and HepG2 cells for the detection of hepatotoxic drugs in humans

Energy Technology Data Exchange (ETDEWEB)

Atienzar, Franck A., E-mail: franck.atienzar@ucb.com [UCB Pharma SA, Non-Clinical Development, Chemin du Foriest, 1420 Braine-l' Alleud (Belgium); Novik, Eric I. [H mu rel Corporation, 675 U.S. Highway 1, North Brunswick, NJ 08902 (United States); Gerets, Helga H. [UCB Pharma SA, Non-Clinical Development, Chemin du Foriest, 1420 Braine-l' Alleud (Belgium); Parekh, Amit [H mu rel Corporation, 675 U.S. Highway 1, North Brunswick, NJ 08902 (United States); Delatour, Claude; Cardenas, Alvaro [UCB Pharma SA, Non-Clinical Development, Chemin du Foriest, 1420 Braine-l' Alleud (Belgium); MacDonald, James [Chrysalis Pharma Consulting, LLC, 385 Route 24, Suite 1G, Chester, NJ 07930 (United States); Yarmush, Martin L. [Department of Biomedical Engineering, Rutgers University, Piscataway, NJ 08854 (United States); Dhalluin, Stéphane [UCB Pharma SA, Non-Clinical Development, Chemin du Foriest, 1420 Braine-l' Alleud (Belgium)

2014-02-15

Drug Induced Liver Injury (DILI) is a major cause of attrition during early and late stage drug development. Consequently, there is a need to develop better in vitro primary hepatocyte models from different species for predicting hepatotoxicity in both animals and humans early in drug development. Dog is often chosen as the non-rodent species for toxicology studies. Unfortunately, dog in vitro models allowing long term cultures are not available. The objective of the present manuscript is to describe the development of a co-culture dog model for predicting hepatotoxic drugs in humans and to compare the predictivity of the canine model along with primary human hepatocytes and HepG2 cells. After rigorous optimization, the dog co-culture model displayed metabolic capacities that were maintained up to 2 weeks which indicates that such model could be also used for long term metabolism studies. Most of the human hepatotoxic drugs were detected with a sensitivity of approximately 80% (n = 40) for the three cellular models. Nevertheless, the specificity was low approximately 40% for the HepG2 cells and hepatocytes compared to 72.7% for the canine model (n = 11). Furthermore, the dog co-culture model showed a higher superiority for the classification of 5 pairs of close structural analogs with different DILI concerns in comparison to both human cellular models. Finally, the reproducibility of the canine system was also satisfactory with a coefficient of correlation of 75.2% (n = 14). Overall, the present manuscript indicates that the dog co-culture model may represent a relevant tool to perform chronic hepatotoxicity and metabolism studies. - Highlights: • Importance of species differences in drug development. • Relevance of dog co-culture model for metabolism and toxicology studies. • Hepatotoxicity: higher predictivity of dog co-culture vs HepG2 and human hepatocytes.

Primary chondrocytes enhance cartilage tissue formation upon co-culture with expanded chondrocytes, dermal fibroblasts, 3T3 feeder cells and embryonic stem cells

NARCIS (Netherlands)

Hendriks, J.A.A.; Miclea, Razvan L.; Schotel, Roka; de Bruijn, Ewart; Moroni, Lorenzo; Karperien, Hermanus Bernardus Johannes; Riesle, J.U.; van Blitterswijk, Clemens

2010-01-01

Co-culture models have been increasingly used in tissue engineering applications to understand cell–cell interactions and consequently improve regenerative medicine strategies. Aiming at further elucidating cartilage tissue formation, we co-cultured bovine primary chondrocytes (BPCs) with human

Permeability of PEGylated immunoarsonoliposomes through in vitro blood brain barrier-medulloblastoma co-culture models for brain tumor therapy.

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Al-Shehri, Abdulghani; Favretto, Marco E; Ioannou, Panayiotis V; Romero, Ignacio A; Couraud, Pierre-Olivier; Weksler, Babette Barbash; Parker, Terry L; Kallinteri, Paraskevi

2015-03-01

Owing to restricted access of pharmacological agents into the brain due to blood brain barrier (BBB) there is a need: 1. to develop a more representative 3-D-co-culture model of tumor-BBB interaction to investigate drug and nanoparticle transport into the brain for diagnostic and therapeutic evaluation. 2. to address the lack of new alternative methods to animal testing according to replacement-reduction-refinement principles. In this work, in vitro BBB-medulloblastoma 3-D-co-culture models were established using immortalized human primary brain endothelial cells (hCMEC/D3). hCMEC/D3 cells were cultured in presence and in absence of two human medulloblastoma cell lines on Transwell membranes. In vitro models were characterized for BBB formation, zonula occludens-1 expression and permeability to dextran. Transferrin receptors (Tfr) expressed on hCMEC/D3 were exploited to facilitate arsonoliposome (ARL) permeability through the BBB to the tumor by covalently attaching an antibody specific to human Tfr. The effect of anticancer ARLs on hCMEC/D3 was assessed. In vitro BBB and BBB-tumor co-culture models were established successfully. BBB permeability was affected by the presence of tumor aggregates as suggested by increased permeability of ARLs. There was a 6-fold and 8-fold increase in anti-Tfr-ARL uptake into VC312R and BBB-DAOY co-culture models, respectively, compared to plain ARLs. The three-dimensional models might be appropriate models to study the transport of various drugs and nanocarriers (liposomes and immunoarsonoliposomes) through the healthy and diseased BBB. The immunoarsonoliposomes can be potentially used as anticancer agents due to good tolerance of the in vitro BBB model to their toxic effect.

Expression of human immunodeficiency virus (HIV) in naturally infected peripheral blood mononuclear cells: comparison of a standard co-culture technique with a newly developed microculture method.

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Eberlein, B; Baur, A; Neundorfer, M; Jahn, G

1991-05-01

Peripheral blood mononuclear cells (PBMCs) from 29 patients infected with human immunodeficiency virus (HIV) were cultured by two different methods. One was the standard co-culture technique, the other a newly developed microculture method. In this assay 10(6) PBMCs were cultivated in 250 microliters medium, no activating agents or allogeneic cells were present. P24 antigen production measured by this method was found in 7 out of 11 PBMC cultures of patients in the Walter Reed (WR) stage 1 or 2, whereas only 4 samples were positive by the co-culture procedure. Cultures from patients in the later stages of the disease (WR 5/6) showed a higher p24 production by the co-culture method than by the microculture assay. It is assumed that rapidly growing HIV strains can be better assessed by the co-culture method which may select for these strains. P24 expression can be more easily obtained by the microculture technique even in cases where slowly replicating strains may be present. In conclusion, results from the microculture procedure described may be a useful supplementation to findings observed by the co-culture method.

Human induced pluripotent stem cell (hiPSC)-derived neurons respond to convulsant drugs when co-cultured with hiPSC-derived astrocytes.

Science.gov (United States)

Ishii, Misawa Niki; Yamamoto, Koji; Shoji, Masanobu; Asami, Asano; Kawamata, Yuji

2017-08-15

Accurate risk assessment for drug-induced seizure is expected to be performed before entering clinical studies because of its severity and fatal damage to drug development. Induced pluripotent stem cell (iPSC) technology has allowed the use of human neurons and glial cells in toxicology studies. Recently, several studies showed the advantage of co-culture system of human iPSC (hiPSC)-derived neurons with rodent/human primary astrocytes regarding neuronal functions. However, the application of hiPSC-derived neurons for seizure risk assessment has not yet been fully addressed, and not at all when co-cultured with hiPSC-derived astrocytes. Here, we characterized hiPSC-derived neurons co-cultured with hiPSC-derived astrocytes to discuss how hiPSC-derived neurons are useful to assess seizure risk of drugs. First, we detected the frequency of spikes and synchronized bursts hiPSC-derived neurons when co-cultured with hiPSC-derived astrocytes for 8 weeks. This synchronized burst was suppressed by the treatment with 6-cyano-7-nitroquinoxaline-2,3-dione, α-amino-3-hydroxy-5-methylisoxazole-4-propionic acid (AMPA) receptor antagonist, and D-(-)-2-amino-5-phosphonopentanoic acid, an N-Methyl-d-aspartate (NMDA) receptor antagonist. These data suggested that co-cultured hiPSC-derived neurons formed synaptic connections mediated by AMPA and NMDA receptors. We also demonstrated that co-cultured hiPSC-derived neurons showed epileptiform activity upon treatment with gabazine or kaliotoxin. Finally, we performed single-cell transcriptome analysis in hiPSC-derived neurons and found that hiPSC-derived astrocytes activated the pathways involved in the activities of AMPA and NMDA receptor functions, neuronal polarity, and axon guidance in hiPSC-derived neurons. These data suggested that hiPSC-derived astrocytes promoted the development of action potential, synaptic functions, and neuronal networks in hiPSC-derived neurons, and then these functional alterations result in the epileptiform

A pilot-scale study of biohydrogen production from distillery effluent using defined bacterial co-culture

Energy Technology Data Exchange (ETDEWEB)

Vatsala, T.M.; Raj, S. Mohan; Manimaran, A. (Shri AMM Murugappa Chettiar Research Centre, Photosynthesis and Energy Division, Tharamani, Chennai, India, 600)

2008-10-15

We evaluated the feasibility of improving the scale of hydrogen (H{sub 2}) production from sugar cane distillery effluent using co-cultures of Citrobacter freundii 01, Enterobacter aerogenes E10 and Rhodopseudomonas palustris P2 at 100 m{sup 3} scale. The culture conditions at 100 ml and 2 L scales were optimized in minimal medium and we observed that the co-culture of the above three strains enhanced H{sub 2} productivity significantly. Results at the 100 m{sup 3} scale revealed a maximum of 21.38 kg of H{sub 2}, corresponding to 10692.6 mol, which was obtained through batch method at 40 h from reducing sugar (3862.3 mol) as glucose. The average yield of H{sub 2} was 2.76 mol mol{sup -1} glucose, and the rate of H{sub 2} production was estimated as 0.53 kg/100 m{sup 3}/h. Our results demonstrate the utility of distillery effluent as a source of clean alternative energy and provide insights into treatment for industrial exploitation. (author)

3D printed scaffolds of calcium silicate-doped β-TCP synergize with co-cultured endothelial and stromal cells to promote vascularization and bone formation.

Science.gov (United States)

Deng, Yuan; Jiang, Chuan; Li, Cuidi; Li, Tao; Peng, Mingzheng; Wang, Jinwu; Dai, Kerong

2017-07-17

Synthetic bone scaffolds have potential application in repairing large bone defects, however, inefficient vascularization after implantation remains the major issue of graft failure. Herein, porous β-tricalcium phosphate (β-TCP) scaffolds with calcium silicate (CS) were 3D printed, and pre-seeded with co-cultured human umbilical cord vein endothelial cells (HUVECs) and human bone marrow stromal cells (hBMSCs) to construct tissue engineering scaffolds with accelerated vascularization and better bone formation. Results showed that in vitro β-TCP scaffolds doped with 5% CS (5%CS/β-TCP) were biocompatible, and stimulated angiogenesis and osteogenesis. The results also showed that 5%CS/β-TCP scaffolds not only stimulated co-cultured cells angiogenesis on Matrigel, but also stimulated co-cultured cells to form microcapillary-like structures on scaffolds, and promoted migration of BMSCs by stimulating co-cultured cells to secrete PDGF-BB and CXCL12 into the surrounding environment. Moreover, 5%CS/β-TCP scaffolds enhanced vascularization and osteoinduction in comparison with β-TCP, and synergized with co-cultured cells to further increase early vessel formation, which was accompanied by earlier and better ectopic bone formation when implanted subcutaneously in nude mice. Thus, our findings suggest that porous 5%CS/β-TCP scaffolds seeded with co-cultured cells provide new strategy for accelerating tissue engineering scaffolds vascularization and osteogenesis, and show potential as treatment for large bone defects.

Intake, selection, digesta retention, digestion and gut fill of two coprophageous species, rabbits (Oryctolagus cuniculus) and guinea pigs (Cavia porcellus), on a hay-only diet.

Science.gov (United States)

Franz, R; Kreuzer, M; Hummel, J; Hatt, J-M; Clauss, M

2011-10-01

A colonic separation mechanism (CSM) is the prerequisite for the digestive strategy of coprophagy. Two different CSM are known in small herbivores, the 'wash-back' CSM of lagomorphs and the 'mucous-trap' CSM of rodents. Differences between these groups in their digestive pattern when fed exclusively hay were investigated in six rabbits (Oryctolagus cuniculus) and six guinea pigs (Cavia porcellus). Intake, digestibility (by total faecal collection), solute and particle mean retention times (MRT, using Co-EDTA and Cr-mordanted fibres) were measured. Rabbits selected less fibrous parts of the hay than guinea pigs, leaving orts with higher content of neutral detergent fibre [NDF; 721 ± 21 vs. 642 ± 31 g/kg dry matter (DM) in guinea pigs]. They also expressed a lower NDF digestibility (0.44 ± 0.10 vs. 0.55 ± 0.05 of total), a similar particle MRT (15 ± 3 vs. 18 ± 6 h), a longer solute MRT (51 ± 9 vs. 16 ± 4 h), and a lower calculated dry matter gut fill (19.6 ± 4.7 vs. 29.7 ± 4.1 g DM/kg body mass) than guinea pigs (p bacterial matter from the colonic digesta plug than the 'mucous-trap' CSM found in the guinea pigs. Related to metabolic body mass, rabbits therefore need a less capacious colon for their CSM where a more efficient bacteria wash-out is reflected in the lower fibre digestibility. A lighter digestive tract could contribute to a peculiarity of lagomorphs: their ability to run faster than other similar-sized mammals. © 2010 Blackwell Verlag GmbH.

Co-Metabolic Degradation of β-Cypermethrin and 3-Phenoxybenzoic Acid by Co-Culture of Bacillus licheniformis B-1 and Aspergillus oryzae M-4.

Science.gov (United States)

Zhao, Jiayuan; Chi, Yuanlong; Xu, Yingchao; Jia, Dongying; Yao, Kai

2016-01-01

The degradation efficiency of organic contaminants and their associated metabolites by co-culture of microbes is mainly limited by toxic intermediates from co-metabolic degradation. In this study, we investigated the degradation of β-cypermethrin (β-CY) and 3-phenoxybenzoic acid (3-PBA) by co-culture of Bacillus licheniformis B-1 and Aspergillus oryzae M-4, as well as the influences of β-CY and 3-PBA metabolites on their degradation and the growth of strains B-1 and M-4. Our results indicated that 100 mg/L β-CY was degraded by 78.85%, and 3-PBA concentration was 0.05 mg/L after 72 h. Compared with using only strain B-1, the half-life (t1/2) of β-CY by using the two strains together was shortened from 84.53 h to 38.54 h, and the yield coefficient of 3-PBA was decreased from 0.846 to 0.001. At 100 mg/L of 3-PBA and gallic acid, β-CY and 3-PBA degradation were only 17.68% and 40.45%, respectively. As the toxic intermediate derived from co-metabolic degradation of β-CY by strain B-1, 3-PBA was efficiently degraded by strain M-4, and gallic acid, as the toxic intermediate from co-metabolic degradation of 3-PBA by strain M-4, was efficiently degraded by strain B-1. These results provided a promising approach for efficient biodegradation of β-CY and 3-PBA.

Co-Metabolic Degradation of β-Cypermethrin and 3-Phenoxybenzoic Acid by Co-Culture of Bacillus licheniformis B-1 and Aspergillus oryzae M-4.

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Jiayuan Zhao

Full Text Available The degradation efficiency of organic contaminants and their associated metabolites by co-culture of microbes is mainly limited by toxic intermediates from co-metabolic degradation. In this study, we investigated the degradation of β-cypermethrin (β-CY and 3-phenoxybenzoic acid (3-PBA by co-culture of Bacillus licheniformis B-1 and Aspergillus oryzae M-4, as well as the influences of β-CY and 3-PBA metabolites on their degradation and the growth of strains B-1 and M-4. Our results indicated that 100 mg/L β-CY was degraded by 78.85%, and 3-PBA concentration was 0.05 mg/L after 72 h. Compared with using only strain B-1, the half-life (t1/2 of β-CY by using the two strains together was shortened from 84.53 h to 38.54 h, and the yield coefficient of 3-PBA was decreased from 0.846 to 0.001. At 100 mg/L of 3-PBA and gallic acid, β-CY and 3-PBA degradation were only 17.68% and 40.45%, respectively. As the toxic intermediate derived from co-metabolic degradation of β-CY by strain B-1, 3-PBA was efficiently degraded by strain M-4, and gallic acid, as the toxic intermediate from co-metabolic degradation of 3-PBA by strain M-4, was efficiently degraded by strain B-1. These results provided a promising approach for efficient biodegradation of β-CY and 3-PBA.

Effects of Treatment with Bone Morphogenetic Protein 4 and Co-culture on Expression of Piwil2 Gene in Mouse Differentiated Embryonic Stem Cells

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Mehdi Forouzandeh-Moghadam

2009-01-01

Full Text Available Background: Specific growth factors and feeder layers seem to have important roles in in vitroembryonic stem cells (ESCs differentiation. In this study,the effects of bone morphogenetic protein4 (BMP4 and mouse embryonic fibroblasts (MEFs co-culture system on germ cell differentiationfrom mouse ESCs were studied.Materials and Methods: Cell suspension was prepared from one-day-old embryoid body (EBand cultured for four days in DMEM medium containing 20% fetal bovine serum (FBS in thefollowing groups: simple culture (SC, simple culture with BMP4 (SCB, co-culture (CO-C andco-culture with BMP4 (CO-CB. Expression of piwi-like homolog 2 (Piwil2, the germ cell-specificgene, was evaluated in the different study groups by using quantitative real time polymerase chainreaction (RT-PCR. Testis was used as a positive control.Results: The maximum and minimum Piwil2 expression was observed in SC and SCB groups,respectively. A significant difference was observed in Piwil2 expression between SCB and otherstudy groups (p<0.05.Conclusion: The findings of this study showed that neither the addition of BMP4 in culture mediumnor the use of MEFs as a feeder layer have a positive effect on late germ cell induction from mouseESCs.

Review of vascularised bone tissue-engineering strategies with a focus on co-culture systems.

Science.gov (United States)

Liu, Yuchun; Chan, Jerry K Y; Teoh, Swee-Hin

2015-02-01

Poor angiogenesis within tissue-engineered grafts has been identified as a main challenge limiting the clinical introduction of bone tissue-engineering (BTE) approaches for the repair of large bone defects. Thick BTE grafts often exhibit poor cellular viability particularly at the core, leading to graft failure and lack of integration with host tissues. Various BTE approaches have been explored for improving vascularisation in tissue-engineered constructs and are briefly discussed in this review. Recent investigations relating to co-culture systems of endothelial and osteoblast-like cells have shown evidence of BTE efficacy in increasing vascularization in thick constructs. This review provides an overview of key concepts related to bone formation and then focuses on the current state of engineered vascularized co-culture systems using bone repair as a model. It will also address key questions regarding the generation of clinically relevant vascularized bone constructs as well as potential directions and considerations for research with the objective of pursuing engineered co-culture systems in other disciplines of vascularized regenerative medicine. The final objective is to generate serious and functional long-lasting vessels for sustainable angiogenesis that will enable enhanced cellular survival within thick voluminous bone grafts, thereby aiding in bone formation and remodelling in the long term. However, more evidence about the quality of blood vessels formed and its associated functional improvement in bone formation as well as a mechanistic understanding of their interactions are necessary for designing better therapeutic strategies for translation to clinical settings. Copyright © 2012 John Wiley & Sons, Ltd.

Differential susceptibility to plasticity: a 'missing link' between gene-culture co-evolution and neuropsychiatric spectrum disorders?

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Wurzman Rachel

2012-04-01

Full Text Available Abstract Brüne's proposal that erstwhile 'vulnerability' genes need to be reconsidered as 'plasticity' genes, given the potential for certain environments to yield increased positive function in the same domain as potential dysfunction, has implications for psychiatric nosology as well as a more dynamic understanding of the relationship between genes and culture. In addition to validating neuropsychiatric spectrum disorder nosologies by calling for similar methodological shifts in gene-environment-interaction studies, Brüne's position elevates the importance of environmental contexts - inclusive of socio-cultural variables - as mechanisms that contribute to clinical presentation. We assert that when models of susceptibility to plasticity and neuropsychiatric spectrum disorders are concomitantly considered, a new line of inquiry emerges into the co-evolution and co-determination of socio-cultural contexts and endophenotypes. This presents potentially unique opportunities, benefits, challenges, and responsibilities for research and practice in psychiatry. Please see related manuscript: http://www.biomedcentral.com/1741-7015/10/38

Antimicrobial resistance of Aeromonas hydrophila isolated from different food sources: A mini-review

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Deyan Stratev

2016-09-01

Full Text Available Summary: Aeromonas hydrophila is a Gram-negative, oxidase-positive, facultative, anaerobic, opportunistic aquatic pathogen. A. hydrophila produces virulence factors, such as hemolysins, aerolysins, adhesins, enterotoxins, phospholipase and lipase. In addition to isolation from aquatic sources, A. hydrophila has been isolated from meat and meat products, milk and dairy products, and vegetables. However, various studies showed that this opportunistic pathogen is resistant to commercial antibiotics. This is attributed to factors such as the indiscriminate use of antibiotics in aquaculture, plasmids or horizontal gene transfer. In this report, we highlight the occurrence, prevalence and antimicrobial resistance of A. hydrophila isolated from different food samples. The presence of antimicrobial-resistant A. hydrophila in food poses threats to public and aquatic animal health. Keywords: A. hydrophila, Antimicrobial resistance, Microbial food safety

Effect of Bifidobacterium upon Clostridium difficile growth and toxicity when co-cultured in different prebiotic substrates

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Lorena Valdés Varela

2016-05-01

Full Text Available The intestinal overgrowth of Clostridium difficile, often after disturbance of the gut microbiota by antibiotic treatment, leads to C. difficile infection (CDI which manifestation ranges from mild diarrhoea to life-threatening conditions. The increasing CDI incidence, not only in compromised subjects but also in traditionally considered low-risk populations, together with the frequent relapses of the disease, has attracted the interest for prevention/therapeutic options. Among these, probiotics, prebiotics or synbiotics constitute a promising approach. In this study we determined the potential of selected Bifidobacterium strains for the inhibition of C. difficile growth and toxicity in different carbon sources. We conducted co-cultures of the toxigenic strain C. difficile LMG21717 with four Bifidobacterium strains (Bifidobacterium longum IPLA20022, Bifidobacterium breve IPLA20006, Bifidobacterium bifidum IPLA20015, and Bifidobacterium animalis subsp. lactis Bb12 in the presence of various prebiotic substrates (Inulin, Synergy and Actilight or glucose, and compared the results with those obtained for the corresponding mono-cultures. C. difficile and bifidobacteria levels were quantified by qPCR; the pH and the production of short chain fatty acids was also determined. Moreover, supernatants of the cultures were collected to evaluate their toxicity using a recently developed model. Results showed that co-culture with B. longum IPLA20022 and B. breve IPLA20006 in the presence of short-chain fructooligosaccharides, but not of Inulin, as carbon source significantly reduced the growth of the pathogen. With the sole exception of B. animalis Bb12, whose growth was enhanced, the presence of C. difficile did not show major effects upon the growth of the bifidobacteria. In accordance with the growth data, B. longum and B. breve were the strains showing higher reduction in the toxicity of the co-culture supernatants.

Micropatterned co-culture of hepatocyte spheroids layered on non-parenchymal cells to understand heterotypic cellular interactions

International Nuclear Information System (INIS)

Otsuka, Hidenori; Sasaki, Kohei; Okimura, Saya; Nagamura, Masako; Nakasone, Yuichi

2013-01-01

Microfabrication and micropatterning techniques in tissue engineering offer great potential for creating and controlling cellular microenvironments including cell–matrix interactions, soluble stimuli and cell–cell interactions. Here, we present a novel approach to generate layered patterning of hepatocyte spheroids on micropatterned non-parenchymal feeder cells using microfabricated poly(ethylene glycol) (PEG) hydrogels. Micropatterned PEG-hydrogel-treated substrates with two-dimensional arrays of gelatin circular domains (ϕ = 100 μm) were prepared by photolithographic method. Only on the critical structure of PEG hydrogel with perfect protein rejection, hepatocytes were co-cultured with non-parenchymal cells to be led to enhanced hepatocyte functions. Then, we investigated the mechanism of the functional enhancement in co-culture with respect to the contributions of soluble factors and direct cell–cell interactions. In particular, to elucidate the influence of soluble factors on hepatocyte function, hepatocyte spheroids underlaid with fibroblasts (NIH/3T3 mouse fibroblasts) or endothelial cells (BAECs: bovine aortic endothelial cells) were compared with physically separated co-culture of hepatocyte monospheroids with NIH3T3 or BAEC using trans-well culture systems. Our results suggested that direct heterotypic cell-to-cell contact and soluble factors, both of these between hepatocytes and fibroblasts, significantly enhanced hepatocyte functions. In contrast, direct heterotypic cell-to-cell contact between hepatocytes and endothelial cells only contributed to enhance hepatocyte functions. This patterning technique can be a useful experimental tool for applications in basic science, drug screening and tissue engineering, as well as in the design of artificial liver devices. (paper)

Role of differential physical properties in emergent behavior of 3D cell co-cultures

Science.gov (United States)

Kolbman, Dan; Das, Moumita

2015-03-01

The biophysics of binary cell populations is of great interest in many biological processes, whether the formation of embryos or the initiation of tumors. During these processes, cells are surrounded by other cell types with different physical properties, often with important consequences. For example, recent experiments on a co-culture of breast cancer cells and healthy breast epithelial cells suggest that the mechanical mismatch between the two cell types may contribute to enhanced migration of the cancer cells. Here we explore how the differential physical properties of different cell types may influence cell-cell interaction, aggregation, and migration. To this end, we study a proof of concept model- a three-dimensional binary system of interacting, active, and deformable particles with different physical properties such as elastic stiffness, contractility, and particle-particle adhesion, using Langevin Dynamics simulations. Our results may provide insights into emergent behavior such as segregation and differential migration in cell co-cultures in three dimensions.

Influence of epidermal growth factor (EGF) and hydrocortisone on the co-culture of mature adipocytes and endothelial cells for vascularized adipose tissue engineering.

Science.gov (United States)

Huber, Birgit; Czaja, Alina Maria; Kluger, Petra Juliane

2016-05-01

The composition of vascularized adipose tissue is still an ongoing challenge as no culture medium is available to supply adipocytes and endothelial cells appropriately. Endothelial cell medium is typically supplemented with epidermal growth factor (EGF) as well as hydrocortisone (HC). The effect of EGF on adipocytes is discussed controversially. Some studies say it inhibits adipocyte differentiation while others reported of improved adipocyte lipogenesis. HC is known to have lipolytic activities, which might result in mature adipocyte dedifferentiation. In this study, we evaluated the influence of EGF and HC on the co-culture of endothelial cells and mature adipocytes regarding their cell morphology and functionality. We showed in mono-culture that high levels of HC promoted dedifferentiation and proliferation of mature adipocytes, whereas EGF seemed to have no negative influence. Endothelial cells kept their typical cobblestone morphology and showed a proliferation rate comparable to the control independent of EGF and HC concentration. In co-culture, HC promoted dedifferentiation of mature adipocytes, which was shown by a higher glycerol release. EGF had no negative impact on adipocyte morphology. No negative impact on endothelial cell morphology and functionality could be seen with reduced EGF and HC supplementation in co-culture with mature adipocytes. Taken together, our results demonstrate that reduced levels of HC are needed for co-culturing mature adipocytes and endothelial cells. In co-culture, EGF had no influence on mature adipocytes. Therefore, for the composition of vascularized adipose tissue constructs, the media with low levels of HC and high or low levels of EGF can be used. © 2016 International Federation for Cell Biology.

Oxidative damage to DNA by diesel exhaust particle exposure in co-cultures of human lung epithelial cells and macrophages

DEFF Research Database (Denmark)

Jantzen, Kim; Roursgaard, Martin; Madsen, Claus Desler

2012-01-01

Studies in mono-culture of cells have shown that diesel exhaust particles (DEPs) increase the production of reactive oxygen species (ROS) and oxidative stress-related damage to DNA. However, the level of particle-generated genotoxicity may depend on interplay between different cell types, e.g. lung...... treatment with standard reference DEPs, SRM2975 and SRM1650b. The exposure to DEPs did not affect the colony-forming ability of A549 cells in co-culture with THP-1a cells. The DEPs generated DNA strand breaks and oxidatively damaged DNA, measured using the alkaline comet assay as formamidopyrimidine...... relationship between levels of respiration and ROS production. In conclusion, exposure of mono-cultured cells to DEPs generated oxidative stress to DNA, whereas co-cultures with macrophages had lower levels of oxidatively damaged DNA than A549 epithelial cells....

The gene expression profiles of canine mammary cancer cells grown with carcinoma-associated fibroblasts (CAFs as a co-culture in vitro

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Król Magdalena

2012-03-01

Full Text Available Abstract Background It is supposed that fibroblasts present in tumour microenvironment increase cancer invasiveness and its ability to metastasize but the mechanisms have not been clearly defined yet. Thus, the current study was designed to assess changes in gene expression in five various cancer cell lines grown as a co-culture with the carcinoma-associated fibroblasts (CAFs in vitro. Results A carcinoma-associated fibroblast cell line was isolated from a canine mammary cancer. Then, a co-culture of cancer cells with the CAFs was established and maintained for 72 hrs. Having sorted the cells, a global gene expression in cancer cells using DNA microarrays was examined. The analysis revealed an up-regulation of 100 genes and a down-regulation of 106 genes in the cancer cells grown as a co-culture with the CAFs in comparison to control conditions. The PANTHER binomial statistics tool was applied to determine statistically over-manifested pathways (p Conclusion The results of the current study showed that the co-culturing of cancer cells and the CAFs caused significant changes to the cancer gene expression. The presence of the CAFs in a microenvironment of cancer cells promotes adhesion, angiogenesis and EMT.

Crystallization and initial X-ray analysis of polyhydroxyalkanoate granule-associated protein from Aeromonas hydrophila

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Zhao, Minglian; Li, Zhenguo; Zheng, Wei; Lou, Zhiyong [MOE Key Laboratory of Protein Science, Department of Biological Sciences and Biotechnology, Tsinghua University, Beijing 100084 (China); Chen, Guo-Qiang, E-mail: chengq@stu.edu.cn [MOE Key Laboratory of Protein Science, Department of Biological Sciences and Biotechnology, Tsinghua University, Beijing 100084 (China); Multidisciplinary Research Center, Shantou University, Shantou 515063, Guangdong (China)

2006-08-01

The phasin PhaP{sub Ah} from A. hydrophila strain 4AK4 was crystallized using the hanging-drop vapour-diffusion method. Polyhydroxyalkanoate (PHA) granule-associated proteins (phasins) were discovered in PHA-accumulating bacteria. They play a crucial role as a structural protein during initial PHA-granule formation and granule growth and also serve as interfaces for granule stabilization in vivo. The phasin PhaP{sub Ah} from Aeromonas hydrophila strain 4AK4 was crystallized using the hanging-drop vapour-diffusion method. Single crystals were cryocooled for X-ray diffraction analysis. The phasin crystals belonged to space group P2{sub 1}2{sub 1}2{sub 1}, with unit-cell parameters a = 80.8, b = 108.9, c = 134.4 Å.