Fluid Thioglycollate Research Articles

Perkinsus beihaiensis (Perkinsozoa) in oysters of Bahia State, Brazil.

This study reports the pathogen Perkinsus beihaiensis in oysters of the genus Crassostrea on the coast of the State of Bahia (Brazil), its prevalence, infection intensity and correlation with salinity. Oysters (n = 240) were collected between October and December 2014 at eight sampling stations between latitudes 13°55'S and 15°42'S. The laboratory procedures included macroscopic analysis, histology, culture in Ray's fluid thioglycollate medium (RFTM), Polymerase Chain Reaction (PCR) and DNA sequencing. PCR and sequencing have been used for the genetic identification of oysters as well. Two species of oysters have been identified: Crassostrea rhizophorae and C. brasiliana. In both oyster species P. beihaiensis was the only Perkinsus species detected. In C. rhizophorae, the average prevalence was 82.8% by histology and 65.2% by RFTM. In C. brasiliana, the prevalences were 70.5% and 35.7%, respectively. The higher prevalence of P. beihaiensis in C. rhizophorae was probably influenced by salinity, with which was positively correlated (r> 0.8). In both oysters, P. beihaiensis was located mainly in the gastric epithelium. The infection was generally mild or moderate, without apparent harm to the hosts, but in cases of severe infection, there was hemocytical reaction and tissue disorganization. The generally high prevalence in the region suggests that oysters should be monitored with respect to this pathogen, especially in growing areas.

Genetic signature analysis of Perkinsus marinus in Mexico suggests possible translocation from the Atlantic Ocean to the Pacific coast of Mexico

BackgroundThe protozoan Perkinsus marinus (Mackin, Owen & Collier) Levine, 1978 causes perkinsosis in the American oyster Crassostrea virginica Gmelin, 1791. This pathogen is present in cultured C. virginica from the Gulf of Mexico and has been reported recently in Saccostrea palmula (Carpenter, 1857), Crassostrea corteziensis (Hertlein, 1951) and Crassostrea gigas (Thunberg, 1793) from the Mexican Pacific coast. Transportation of fresh oysters for human consumption and repopulation could be implicated in the transmission and dissemination of this parasite across the Mexican Pacific coast. The aim of this study was two-fold. First, we evaluated the P. marinus infection parameters by PCR and RFTM (Ray’s fluid thioglycollate medium) in C. virginica from four major lagoons (Términos Lagoon, Campeche; Carmen-Pajonal-Machona Lagoon complex, Tabasco; Mandinga Lagoon, Veracruz; and La Pesca Lagoon, Tamaulipas) from the Gulf of Mexico. Secondly, we used DNA sequence analyses of the ribosomal non-transcribed spacer (rNTS) region of P. marinus to determine the possible translocation of this species from the Gulf of Mexico to the Mexican Pacific coast.ResultsPerkinsus marinus prevalence by PCR was 57.7% (338 out of 586 oysters) and 38.2% (224 out of 586 oysters) by RFTM. The highest prevalence was observed in the Carmen-Pajonal-Machona Lagoon complex in the state of Tabasco (73% by PCR and 58% by RFTM) and the estimated weighted prevalence (WP) was less than 1.0 in the four lagoons. Ten unique rDNA-NTS sequences of P. marinus [termed herein the “P. marinus (Pm) haplotype”] were identified in the Gulf of Mexico sample. They shared 96–100% similarity with 18 rDNA-NTS sequences from the GenBank database which were derived from 16 Mexican Pacific coast infections and two sequences from the USA. The phylogenetic tree and the haplotype network showed that the P. marinus rDNA-NTS sequences from Mexico were distant from the rDNA-NTS sequences of P. marinus reported from the USA. The ten rDNA-NTS sequences described herein were restricted to specific locations displaying different geographical connections within the Gulf of Mexico; the Carmen-Pajonal-Machona Pm1 haplotype from the state of Tabasco shared a cluster with P. marinus isolates reported from the Mexican Pacific coast.ConclusionsThe rDNA-NTS sequences of P. marinus from the state of Tabasco shared high similarity with the reference rDNA-NTS sequences from the Mexican Pacific coast. The high similarity suggests a transfer of oysters infected with P. marinus from the Mexican part of the Gulf of Mexico into the Mexican Pacific coast.

EMERGENCE AND ANTIBIOTIC SENSITIVITY PATTERN OF ACINETOBACTER BAUMANNII IN HOSPITAL FACILITY

ABSTRACTObjective: Nosocomial infections or Hospital acquired infection (HAI) are one of the major threats to hospitalized patients as well as for the hospitalassociated personnel. In last few years there is a gross change in causative agents, new organisms have come out with great threat to hospitals as theypossess antibiotic resistance property e.g. production of biofilm, production of enzymes such as β- lactamases. Among many organisms, Acinetobacterbaumannii has emerged as a potent nosocomial pathogen. Our objective of this study was to find the burden of Acinetobacter baumannii infectionswhich are associated as nosocomial infections and to determine the drug of choice for an effective treatment.Methods: Clinical specimens were collected from patients of different unit of the hospital by maintaining universal precautions and standardmicrobiological protocols. All the respective specimens were cultured in respective culture medium i.e. MacConkey agar, blood agar, chocolate agar,cysteine lactose electrolyte deficient (CLED) agar and, fluid thioglycolate (TG) medium at 37˚C for 24-48 hours. After incubation of 24-48 hours cultureplates were examined for bacterial growth and identification and antibiotic sensitivity test was made by Vitek2 compact.Result: The study was conducted at the department of microbiology from January 2016 to April 2016. A total of 2582 specimens were collected andprocessed for identification and sensitivity testing. Specimens of all age group (2 days- 93 years) and both sexes were processed for identificationof A. baumannii and antibiotic sensitivity testing. A total of 119 isolates (4.60%) of A. baumannii were obtained from 2582 clinical specimens. Themost common infection A. baumannii was found as lower respiratory tract infection (89.07%) followed by abscess (6.72%), septicaemia (2.52%),urinary tract infections (0.84%), and soft tissue infections (0.84%). The maximum sensitivity of A. baumannii isolates were seen to Colistin (CL) (119,100%), followed by Tigecycline (TGC) (63, 52.94%) and Minocycline (MIN) (27, 22.69%). The maximum resistant was observed for Imipenem (IMI),Aztreonam (AZT) and Ticarcillin- clavulanic acid (TIC) (119, 100%).Conclusion: The Gram- negative coccobacillus, Acinetobacter baumannii poses a formidable threat to patients. It has emerged as a superbug inhospital environment particularly in ICU units. The chances of A. baumannii infections increase in the presence of iatrogenic factors like inadequatelong- term antibiotic therapy and new interventions in a medical facility. To control the burden of Acinetobacter infections new therapies suchas combine therapy must be obtained and followed with proper dose as recommend by physicians; along with awareness of the importance ofthis infection should be implicated. Proper sanitation, good housekeeping, sterilization of equipment, hand hygiene, water purification, isolationprocedures and maintaining of the hospital environment, use of infection control practices are some of the measures to control the transmission ofAcinetobacter spp. among hospital personnel.Keywords: Acinetobacter baumannii, Biofilm, β-lactamases, Hospital acquired infection.

Survey on <i>Perkinsus</i> Species in Manila Clam <i>Ruditapes philippinarum</i> in Korean Waters Using Species-Specific PCR

We investigated possible infection with Perkinsus olseni and/or P. honshuensis in Manila clams in Korean waters using Perkinsus species-specific PCR. Ray’s fluid thioglycollate medium assay revealed that 97.3% of the clams examined were infected with Perkinsus sp., with the mean intensity ranging from 2.2 × 104 to 6.8 × 106 cells/g gill. In the Perkinsus species-specific PCR assay, all the amplified Perkinsus-positive bands were identified as P. olseni, while none of P. honshuensis-specific band was amplified. Our survey results indicate that P. olseni is the main causative agent of perkinsosis in Manila clam populations in Korean waters, while more extensive survey must be carried out.

Novel biorelevant dissolution medium as a prognostic tool for polysaccharide-based colon-targeted drug delivery system.

To overcome the limitations of the conventionally used methods for evaluation of orally administered colon-targeted delivery systems, a novel dissolution method using probiotics has been recently reported. In the present study, universal suitability of this medium composed of five different probiotics is established. Different delivery systems – mini tablets, liquisolid compacts, and microspheres coated with different polysaccharides – were prepared and subjected to sequential dissolution testing in medium with and without microbiota. The results obtained from fluid thioglycollate medium (FTM)-based probiotic medium for all the polysaccharide-based formulations showed statistically similar dissolution profile to that in the rat and goat cecal content media. Hence, it can be concluded that the developed FTM-based probiotic medium, once established, may eliminate the need for further animal sacrifice in the dissolution testing of polysaccharide-based colon-targeted delivery system.

Ammonia-Acetylene PECVD Coating on Wound Dressing to Control Delivery of Silver Sulfadiazine

A novel wound dressing surface modification was developed by applying plasma-enhanced chemical vapor deposition (PECVD). The cotton wound gauze dressings were treated by the mixed composition of 10 %NH3+He and 10 %NH3+C2H2 plasmas followed by immersion in silver sulfadiazine (AgSD) solution. A high functionality and a high surface area can thus be obtained by plasma deposition of C2H2 + NH3. We reported the effect of a strong interaction between AgSD and cotton gauze yarn by observing the reactive amorphous carbon layer. It had an ability to slow down the release rate of AgSD compounds into the solution, reported as the UV-visible absorbance of the solution released from the plasma-treated gauze. The results showed that the release rate of AgSD molecules was lower, compared to the untreated gauze. Scanning electron microscopy (SEM) micrographs showed the interaction between AgSD and the cotton fibers of the plasma-treated gauze. In the untreated gauze, no such interaction was found. According to the energy dispersive X-ray spectroscopy (EDS) data, %weight of the Ag compound on the treated gauze was higher than the untreated gauze. A test on the antibacterial effect, using Fluid Thioglycollate Medium (FTM) as the culture media, clearly demonstrated that the plasma treated gauze which was immersed with AgSD, had ability to inhibit bacterial growth.

Primer registro de Perkinsus sp. (Protozoa, Apicomplexa) en el callo de hacha Atrina maura en Sinaloa, México

We evaluated the presence of the protozoan Perkinsus sp. by determining of its prevalence and level of infection in Atrina maura during 2012. Pen shells were collected per season beside an oyster farm located at Estero La Piedra, Sinaloa, Mexico, and analyzed by Ray's fluid thioglycollate medium and polymerase chain reaction. The higher prevalence (90%) and infection level (2.3) were obtained when temperature and salinity were higher. The Mackin scale showed a non-infected to moderate infection range. PCR was positive to Perkinsus sp. This is the first record of the presence of Perkinsus sp. in A. maura in Sinaloa, Mexico.

First record of Perkinsus chesapeaki infecting Crassostrea rhizophorae in South America

This study investigated Perkinsus spp. infecting Crassostrea rhizophorae from the Jaguaribe River estuary, Ceará, Brazil. Fragments of gills and rectum of the oysters (n=150) were incubated in Ray’s fluid thioglycollate medium (RFTM). Genus Perkinsus-specific PerkITS85/750 PCR assays were performed and their amplicons were sequenced by the Sanger method. The RFTM assays confirmed Perkinsus spp. The sequencing of the amplified fragments from the rDNA internal transcribed spacers (ITS) of Perkinsus spp. confirmed Perkinsus chesapeaki. Neighbor-Joining analyzes place P. chesapeaki identified in this study in a well-supported clade with other isolates of the same species. This is the first record of P. chesapeaki infecting C. rhizophorae in South America.

Perkinsus beihaiensis infectando a ostra Crassostrea rhizophorae em cultivo e em estoque natural na Baía de Camamu, Bahia, Brasil

Foi investigada a infecção de Perkinsus beihaiensis (Perkinsozoa) na ostra Crassostrea rhizophorae em um sistema de cultivo do tipo espinhel e em um estoque natural de ostras no manguezal adjacente ambos localizados no estado da Bahia, Nordeste do Brasil. As colheitas foram realizadas em outubro e novembro de 2012 e em janeiro de 2013. As ostras (n = 300) foram medidas, examinadas macroscopicamente quanto a sinais da infecção e submetidas às técnicas laboratoriais: histologia, ensaio em meio de cultivo de tioglicolato de Ray (RFTM), reação em cadeia da polimerase (PCR) e sequenciamento, que confirmou a identificação do patógeno. As análises histológicas e o RFTM mostraram, respectivamente, prevalência média de 93,3% e de 69%. A infecção foi geralmente leve ou muito leve. Não houve diferença significativa (p > 0,05) entre os ambientes em termos de prevalência ou severidade da infecção. Este é o primeiro registro de P. beihaiensis no estado da Bahia e o segundo em ostras do Brasil e América do Sul.

Epizootiology of Perkinsus sp. inCrassostrea gasar oysters in polyculture with shrimps in northeastern Brazil.

Bivalve culture is of considerable economic and social interest in northeastern (NE) Brazil. The polyculture is an alternative approach to traditional monoculture for reducing the environmental impact of shrimp farming and improving oyster culture. Perkinsus marinus and Perkinsus olseni were found infecting oysters in NE Brazil and can threaten oyster production. This study evaluated Perkinsus spp. occurrence in Crassostrea gasar during all production stages. Oyster spats were produced in a hatchery and grown in shrimp ponds in Rio Grande do Norte state. Perkinsus spp. were surveyed by Ray's fluid thioglycollate medium and confirmed by polymerase chain reaction. Prevalence and intensity of infection were determined in oysters until they reached 7 cm. Results showed that the broodstock was already infected by Perkinsus (60%), but the derived spats were Perkinsus-free. Oyster spats acquired Perkinsus infection when transferred to ponds. The prevalence gradually increased in the seven months following placement in ponds (73%), and then decreased to 17% by the tenth month. The infections were initially mild, but intensity increased at the final growth stage. In conclusion, it is possible to produce Perkinsus-free C. gasar oyster spats from infected broodstock, and their culture in shrimp ponds is feasible.

Evaluation of the effectiveness of manual and automated dialyzers reprocessing after multiple reuses

A cross-sectional study was conducted to evaluate the effectiveness of manual and automated dialyzer reprocessing. Dialyzers were filled with fluid thioglycollate medium from blood and dialysate chambers after being reprocessed and chemically sterilized with 0.2% peracetic acid. They were incubated for 14 days at 35°C ± 2°C, and microbiologic analysis was performed. Microorganisms were identified in 3 of the 11 samples (27.3%) from the blood chambers: Sphingomonas paucimobilis (2/3) and Penicillium spp (1/3) and in 11 of the 11 samples (100%) from the dialysate chambers: S paucimobilis (7/11), Stenotrophomonas maltophilia (4/11), Pseudomonas aeruginosa (3/11), Candida spp (1/11), and Acinetobacter baumannii (1/11). Of the 4 manually reprocessed dialyzers, gram-positive bacillus were identified in 1 sample (25%) from the blood chamber, and Bacillus spp and Burkholderia spp were identified in 1 sample (25%) from the dialysate chamber. The dialyzers reprocessing can pose risks safety because of exposure patient to microorganisms.

An Assessment of Microbiological Methods to Test Sterility of Foot-and-mouth Disease Vaccines Produced in Brazil

Background: Foot-and-mouth vaccines are an important tool in the control and eradication of the disease. In order to be commercialized, vaccines produced in Brazil undergo an evaluation process by health authorities, which includes sterility testing, residual active virus, potency, thermal stability, volume and non-structural protein activity. Sterility tests described in the Brazilian Pharmacopeia and by the World Organization for Animal Health (OIE) include direct inoculation and membrane filtration methods. The objective of the present study was to evaluate these two methods used to analyze sterility of vaccines against foot-and-mouth disease produced in Brazil.Materials, Methods & Results: Vaccines produced by the six main laboratories in Brazil were initially tested for filtration capacity. The sensitivity of the two techniques was determined artificially contaminating vaccines using known bacterial concentrations. Vaccines (9 bottles) from the same manufacturer were inoculated with 5 mL of steady-state growths of Pseudomonas aeruginosa, Candida albicans, and Clostridium sporogenes to final concentrations of 0.1, 1 and 10 CFU/ mL and a final volume of 55 mL. Bottles were manually shaken for 1 min to complete homogenization of contents. Then, 10 mL of each flask were used in assessment of the direct inoculation method, and 10 mL were used to evaluate the membrane filtration technique. Direct inoculation was carried out inoculating 1 mL of the experimentally contaminated vaccine in five tryptic soy broth (TSB) and fluid thioglycollate medium (FTM) bottles. The membrane filtration technique was carried out filtrating 10 mL of the challenged vaccines in a peristaltic pump system (SteritestTM Pump System), where vaccines were initially solubilized in Triton X-100 to promote filtration. Next, membranes are incubated in TSB and FTM. These use two types of culture medium, tryptic soy broth (TSB) and fluid thioglycollate medium (FTM), with incubation times of 20-25ºC and 30-35ºC, respectively, to detect fungi, yeasts, and aerobic and anaerobic bacteria. The medium is incubated for 14 days, to enable the detection of slow-growth microorganisms that may be in a latent stage or weakened due to the extreme conditions of the production process (like the use of cleaning and disinfection agents, ultraviolet light, and preservers, for instance). All vaccines were effectively filtered in the SteritestTM Pump System. Membrane filtration and direct inoculation presented the same sensitivity to detect yeasts (0.1 CFU/mL) and anaerobic organisms (1 CFU/mL). For the detection of aerobic organisms, membrane filtration was 100 times more sensitive, compared to direct inoculation.Discussion: The specialized literature also reports that, apart from the higher sensitivity, membrane filtration affords to reduce contamination during the procedures, since it is carried out in a closed system. In addition, it is indicated in the analysis of large sample volumes. Moreover, membrane filtration reduces the occurrence of false positive results, since it removes the excess vaccine volume from the culture medium, which may be mistaken for turbidity caused by bacterial growth. In this sense, the membrane filtration technique is more appropriate in the control of vaccine sterility in foot-andmouth disease prevention strategies, and is an interesting tool to improve quality control of the product.

Perkinsus sp. infecting the oyster Crassostrea rhizophorae from estuaries of the septentrional Northeast, Brazil.

The mangrove oyster Crassostrea rhizophorae is an estuarine resource exploited by riverside communities in Northeast Brazil. Despite its socioeconomic importance, studies on the health status of this bivalve are scanty in this region. The purpose of the present study was to investigate the presence of the protozoan Perkinsus sp. in C. rhizophorae collected in August and September 2011 in three estuaries of the septentrional Northeast, Brazil: Jaguaribe (Ceará), Camurupim (Piauí) and Carnaubeiras (Maranhão) (n= 150 specimens/site). The samples were submitted to Ray's fluid thioglycollate medium (RFTM), PCR and histology assays. The RFTM assay revealed spherical, blue or bluish-black hypnospores of the genus Perkinsus in 50 specimens (Jaguaribe= 17.3%, Camurupim= 5.3%, Carnaubeiras= 10.6%). The intensity of the infection ranged from very light (1-10 cells per slide) to severe (more than 40 cells in each of 10 fields of the slide) for Jaguaribe; very light for Camurupim and very light to moderate (at least 40 cells observed in each of 10 fields of the slide) for Carnaubeiras. When submitted to confirmatory PCR analysis, 6 cases were confirmed (Jaguaribe=3, Camurupim=1, Carnaubeiras=2). The histology confirmed 21 cases of infection in specimens from the three estuaries. Although local collectors have reported no mortality in oyster populations that might be attributed to infection by Perkinsus, health surveillance of oyster populations in the septentrional region of Northeast Brazil is advisable.

여름철 산란기에 있어 바지락 번식량의 공간적 변이와 기생 원생생물 Perkinsus olseni 감염이 바지락 번식에 미치는 영향

Spatial variation in the reproductive effort of Manila clam Ruditapes philippinarum is often closely associated with variation in the seawater temperature and food availability, which determines gonad maturity and the quantity of gamates produced during spawning. Previous studies also have reported that severe infection by the protozoan parasite Perkinsus olseni exerts a negative impact on clam reproduction, retarding gonad maturation or decreasing the reproductive effort. In the present study, we investigated impacts of P. olseni infection on the reproductive condition of Manila clam during a spawning season. Histology revealed that 54% of female clams in Wando off the south coast were in spawning, while only 10% of the female from Gomso and 0% of the female from Seonjaedo in Gyeonggi bay off the west coast were engaged in spawning at the end of May in 2004. Ray's fluid thioglycollate media (RFTM) assay was applied to assess P. olseni infection and indicated that the infection intensity in Wando (3,608,000 ± 258,000 cells/g wet tissue) was significantly higher than the levels in Gomso (1,305,000 ± 106,000 cells/g wet

Perkinsus sp. infections and in vitro isolates from Anadara trapezia (mud arks) of Queensland, Australia.

Perkinsus sp. protists were found infecting Anadara trapezia mud ark cockles at 6 sites in Moreton Bay, Queensland, Australia, at prevalences of 4 to 100% during 2011 as determined by surveys using Ray's fluid thioglycollate medium. Perkinsus sp. lesions were found among gill and visceral connective tissues in histological samples from several cockles, where basophilic, eccentrically vacuolated Perkinsus sp. signet ring trophozoites and proliferating, Perkinsus sp. schizont cells were documented. Two Perkinsus sp. isolates were propagated in vitro during August 2013 from gill tissues of a single infected A. trapezia cockle from Wynnum in Moreton Bay. DNA from those isolate cells amplified universally by a Perkinsus genus-specific PCR assay, and rDNA-internal transcribed spacer sequences respectively grouped them with P. olseni and P. chesapeaki in phylogenetic analyses. This is the first report of P. chesapeaki in Australia, and the first report of a P. chesapeaki in vitro isolate from an Australian mollusc host. Although P. olseni was originally described in 1981 as a pathogen of abalone in South Australia, and has subsequently been identified as a prevalent pathogen of numerous other molluscs worldwide, this is also the first report of a P. olseni-like in vitro isolate from an Australian mollusc host.

Microbial flora in chronic periodontitis: study at a tertiary health care center from north karnataka.

Background and Objective:Periodontitis is a major public health problem in India with a prevalence of 60–80%. If untreated it acts as a risk factor for systemic diseases. Data on anaerobic periodontal microflora in the Indian population is very scarce. Hence, this study was undertaken to know the nature of oral microbiota in chronic periodontitis in this region of India and also the semiquantitative study in pre- and post-treatment group and to determine antibiotic susceptibility pattern for aerobic isolates.Materials and Methods:The present study was conducted on 60 cases. Material was collected from the subgingival pockets in patients with chronic periodontitis attending the Periodontology, Outpatient Department. Clinical samples were transported to the laboratory in fluid thioglycollate medium. Initially Gram's stain and Fontana stains were done. Aerobic, anaerobic, and microaerophilic culture were put up. Antibiotic sensitivity test was done for aerobic isolates.Results:Sixty samples yielded 121 isolates of which 78.34% were polymicrobial, 11.66% were monomicrobial and oral commensals were grown in 10% cases. Out of 121 isolates 91.74% were anaerobic, 7.43% were aerobic and 0.83% were microaerophilic. Fusobacterium species was the most common isolate among anaerobes. Using “paired t-test” “P” value was significant indicating significant reduction in colony count after phase-I periodontal therapy.Conclusion:This study has shown that anaerobic bacteria are important cause of chronic periodontitis, along with aerobes and microaerophilic organisms. Fusobacterium spp, Bacteroides fragilis, Porphyromonas spp and Prevotella intermedia are the most common anaerobic pathogens. Bacterial culture methods are still economical and gold standard.

In-house development of an optimized synthetic module for routine [11C]acetate production.

[11C]Acetate, a radiotracer for PET imaging, is a promising radiopharmaceutical for overcoming the limitation of 2-deoxy-2-[18F]fluoro-D-glucose in a number of cancers. Here, the optimized automatic synthesis of [11C]acetate using an in-house-developed module under different conditions has been reported for routine production. [11C]CO2 was produced in a 16.4 MeV PETtrace cyclotron, and methyl magnesium chloride was used for synthesis. For product purification, ion-exchange solid-phase extraction cartridges were used, connected in series. High-performance liquid chromatography and gas chromatography were used to measure radiochemical and chemical purity. The Limulus amebocyte lysate test and the fluid thioglycollate medium test were performed for quality control of [11C]acetate. The total reaction time of [11C]acetate was within 15 min, and the overall decay-corrected radiochemical yield was 84.33±8.85%. Radiochemical purity was greater than 98% when evaluated on an analytical high-performance liquid chromatography system. No endotoxins or anaerobic bacteria were seen on quality control checks. Optimized production of [11C]acetate was achieved by the in-house module. Radiochemical and biological properties of the [11C]acetate produced were appropriate for clinical PET study.

An in Vitro Experimental Study on the Antimicrobial Activity of Silicone Oil against Anaerobic Bacteria

Purpose: To investigate the in vitro antimicrobial activity of silicone oil against anaerobic agents, specifically Propionibacterium acnes, Peptostreptococcus spp., Peptostreptococcus anaerobius, Bacteroides fragilis, Fuobacterium spp., and Clostridium tertium.Method: A 0.5 McFarland turbidity of Propionibacterium acnes, Peptostreptococcus spp., Peptostreptococcus anaerobius, Bacteroides fragilis, Fuobacterium spp., and Clostridium tertium was prepared, and 0.1 mL was inoculated into 0.9 mL of silicone oil. Control inoculations were performed in anaerobic blood agar and fluid thioglycollate medium without silicone oil.Results: Propionibacterium acnes retained their viability on the 3rd day in the presence of silicone oil. In total, 9.7 × 106 colonies were enumerated from 1 mL of silicone oil. After a prolonged incubation of 7 days, the number of colonies observed was 9.2 × 106. The other bacteria disappeared after the 3rd day of incubation in silicone oil.Conclusions: Propionibacterium acnes, which is the most common chronic postoperative endophthalmitis agent, is thought to be resistant to silicone oil.

First report of Perkinsus beihaiensis in wild clams Anomalocardia brasiliana (Bivalvia: Veneridae) in Brazil

This is the first report of Perkinsus sp. (Bivalvia: Veneridae) infecting wild clams of the species Anomalocardia brasiliana in Brazil. The gill lamellae and rectum of 150 specimens of A. brasiliana collected in the Timonha river estuary (Ceará, Northeastern Brazil) in March 2012 were incubated in Ray’s fluid thioglycollate medium (RFTM) for detection of Perkinsus sp. In RFTM, the prevalence of Perkinsus sp. was 14.7% (22/150) and the intensity of infection ranged from very light (1–10 cells across the slide) to light (12–100 cells). The presence of Perkinsus sp. was confirmed by PCR in seven (31.8%) out of 22 RFTM-positive specimens. DNA sequencing confirmed the presence of the genus Perkinsus and the phylogenetic analysis strongly indicated Perkinsus beihaiensis as the species responsible for the infection.

Sterilization of Chrysomya putoria (Insecta: Diptera: Calliphoridae) eggs for use in biotherapy.

Large-scale, quality-controlled laboratory production of fly larvae is needed for biotherapy. The objective of this study was to assess the action of glutaraldehyde on the sterilization of Chrysomya putoria eggs by applying pharmaceutical sterility tests. Egg masses with 0.600 g were divided into three parts of 0.200 g, the eggs were separated using sterile distilled water, and the suspensions obtained were mixed with activated 2% glutaraldehyde solution. After 15-min contact, the suspensions were filtered through Whatman filter paper, and the glutaraldehyde residue obtained in the filtrate was neutralized by rinsing with Tryptone Soy Broth. The treated eggs were placed aseptically on Petri dishes containing gauze moistened with sterile saline solution. About 10% of the sterilized mass was transferred to test tubes containing Tryptone Soy Broth and Fluid Thioglycollate Broth. The tubes were incubated, respectively, at 22.5 and 35.0°C for 14 d to verify egg mass sterility. The plates containing the rest of the eggs (90%) were sealed with plastic film and kept in a climatized chamber at 30°C/d, 28°C per night, 60 ± 10% relative humidity, and under a 12-h light period to assess insect viability and survival. Each experiment was carried out in triplicate using a biological class II safety cabinet. No change in color or turgidity was observed with the agent tested, proving the sterility of the product and that there was no trace of contamination. Forty larvae (in three replications) in the periods of 12, 24, and 48 h after sterilization, when transferred to diet, produced larvae, pupae, and total viability similar to the control (larvae without sterilization). However, for the 72- h treatment, larvae and total viability were significantly lower than for the other treatments. There was no significant difference for the pupal stage. The product tested was shown to be efficacious for use as a sterilizer of C. putoria eggs for all the parameters assessed.