Development and validation of a PCR-based method to differentiate human sex in blood-fed Aedes aegypti (Diptera: Culicidae).

Monitoring mosquito host choice to identify high-risk groups for different vector-borne diseases is important to devise vector control strategies and disease management. The present study was conducted to develop and validate a PCR-based method to identify human sex in blood-fed Aedes aegypti mosquitoes. Several human genes present in both the X and Y chromosomes were screened and diagnostic PCR primers were successfully designed and amplified for the human STS gene. The limit of detection of this PCR assay was carried out on Ae. aegypti fed with human blood up to 5 days (120 hours) post blood-meal under laboratory condition. The efficiency of this PCR assay was evaluated in field-collected Ae. aegypti mosquitoes and compared with other existing methods. The developed PCR primers can successfully amplify and distinguish human sex in mosquitoes up to 72 hours after a blood meal, with an amplified product of 627bp and 298bp for male (XY) and 627bp for female (XX) blood-fed mosquitoes. Further, validation of this assay in field-collected Ae. aegypti mosquitoes revealed that this assay could detect human sex in mosquito blood meal substantially more efficiently (c2 = 4.5, p = 0.034) than other PCR based assay. The newly developed PCR assay highly specific to human DNA and can distinguish male and female DNA for up to 72 hours. This assay can be is used for identifying highrisk groups and extended to other medically important hematophagous insects to assess their role in disease transmission and epidemic preparedness.

Diverse Circular DNA Viral Communities in Blood, Oral, and Fecal Samples of Captive Lemurs.

Few studies have addressed viral diversity in lemurs despite their unique evolutionary history on the island of Madagascar and high risk of extinction. Further, while a large number of studies on animal viromes focus on fecal samples, understanding viral diversity across multiple sample types and seasons can reveal complex viral community structures within and across species. Groups of captive lemurs at the Duke Lemur Center (Durham, NC, USA), a conservation and research center, provide an opportunity to build foundational knowledge on lemur-associated viromes. We sampled individuals from seven lemur species, i.e., collared lemur (Eulemur collaris), crowned lemur (Eulemur coronatus), blue-eyed black lemur (Eulemur flavifrons), ring-tailed lemur (Lemur catta), Coquerel's sifaka (Propithecus coquereli), black-and-white ruffed lemur (Varecia variegata variegata), and red ruffed lemur (Varecia rubra), across two lemur families (Lemuridae, Indriidae). Fecal, blood, and saliva samples were collected from Coquerel's sifaka and black-and-white ruffed lemur individuals across two sampling seasons to diversify virome biogeography and temporal sampling. Using viral metagenomic workflows, the complete genomes of anelloviruses (n = 4), cressdnaviruses (n = 47), caudoviruses (n = 15), inoviruses (n = 34), and microviruses (n = 537) were determined from lemur blood, feces, and saliva. Many virus genomes, especially bacteriophages, identified in this study were present across multiple lemur species. Overall, the work presented here uses a viral metagenomics approach to investigate viral communities inhabiting the blood, oral cavity, and feces of healthy captive lemurs.

The detection of blood, semen and saliva through fabrics: A pilot study.

This study aimed to identify if biological material could be detected on the opposite side to deposition on fabric by commonly used presumptive and/or secondary tests. Additionally, this study aimed to ascertain if there is a difference in the DNA quantity and quality from samples obtained from both sides of the same substrate: cotton, polyester, denim, or combined viscose and polyester swatches. Blood, semen, or saliva (25 µL) was deposited on one side of 5 replicates of each fabric type and left for 24 h. Blood swatches were tested using Hemastix® and the ABACard® HemaTrace® immunoassay, semen swatches were tested using acid phosphatase (AP) reagent, the ABACard® p30® immunoassay and hematoxylin and eosin staining, and saliva swatches were tested using Phadebas® paper and the RSID-Saliva™ immunoassay. Both sides of each swatch were separately wet/dry swabbed and subjected to DNA analysis. Blood was able to be detected on the underside of all fabrics using both tests. Semen was able to be detected on the underside of swatches using the presumptive AP test but not p30®, and sperm was rarely observed. Saliva was able to be detected by RSID-Saliva™ but not Phadebas® paper when the underside of swatches were tested. Across all biological materials, DNA was able to be recovered from the top side of all 60 swatches. For the underside, DNA was able to be recovered from 54 swatches. Of the 6 swatches that DNA was unable to be recovered from, one sample was from semen and the rest were from saliva. This study has demonstrated that DNA and components of interest in forensically relevant biological material can be recovered from the opposite side to where it was originally deposited, and that observing biological material and/or DNA on one side of fabric does not definitively indicate direct deposition on that side.

Effect of Blood Decontamination Procedures on the Microshear Bond Strength of Resin-modified Glass Ionomer Cement to Resin Composite.

OBJECTIVE: To investigate the effect of decontamination procedures on the microshear bond strength (µSBS) of blood-contaminated resin-modified glass ionomer cement (RMGIC) bonded to resin composite (RC). METHODS: Eighty RMGIC disc specimens were allocated into 5 groups (n=16). All groups except Group 2 were contaminated with blood. Group 1 had no decontamination procedure, Group 3 was decontaminated by rinsing, Group 4 was decontaminated by 34% phosphoric acid etching, and Group 5 was decontaminated by 5% sodium hypochlorite application. RMGIC specimens were subsequently bonded with RC using a universal adhesive in self-etch mode. µSBS tests were conducted using a universal testing machine at a crosshead speed of 1 mm/min. Failure mode analysis was conducted on RMGIC fracture surfaces under a scanning electron microscope. RESULTS: µSBS results indicated that Group 4 had the highest mean µSBS value of 6.22 ± 2.14 MPa, while Group 1 had the lowest mean µSBS value of 3.53 ±1.67 MPa. Significant differences were observed in the µSBS of Group 2 with no contamination (p=0.023) and Group 4 with decontamination by phosphoric acid-etching (p=0.003) when compared to Group 1 with blood contamination. No statistically significant differences (p>0.05) were observed between all other groups' µSBS. For all groups, the predominant mode of failure was adhesive failure between the RMGIC-RC interface, with a few mixed failures in RMGIC for Groups 2-5. CONCLUSIONS: Blood contamination before adhesive application significantly reduced the µSBS between RMGIC and RC. Phosphoric acid etching was the most effective blood decontamination procedure to improve the µSBS.

Infectivity of Plasmodium parasites to Aedes aegypti and Anopheles stephensi mosquitoes maintained on blood-free meals of SkitoSnack.

BACKGROUND: Aedes and Anopheles mosquitoes are responsible for tremendous global health burdens from their transmission of pathogens causing malaria, lymphatic filariasis, dengue, and yellow fever. Innovative vector control strategies will help to reduce the prevalence of these diseases. Mass rearing of mosquitoes for research and support of these strategies presently depends on meals of vertebrate blood, which is subject to acquisition, handling, and storage issues. Various blood-free replacements have been formulated for these mosquitoes, but none of these replacements are in wide use, and little is known about their potential impact on competence of the mosquitoes for Plasmodium infection. METHODS: Colonies of Aedes aegypti and Anopheles stephensi were continuously maintained on a blood-free replacement (SkitoSnack; SS) or bovine blood (BB) and monitored for engorgement and hatch rates. Infections of Ae. aegypti and An. stephensi were assessed with Plasmodium gallinaceum and P. falciparum, respectively. RESULTS: Replicate colonies of mosquitoes were maintained on BB or SS for 10 generations of Ae. aegypti and more than 63 generations of An. stephensi. The odds of engorgement by SS- relative to BB-maintained mosquitoes were higher for both Ae. aegypti (OR = 2.6, 95% CI 1.3-5.2) and An. stephensi (OR 2.7, 95% CI 1.4-5.5), while lower odds of hatching were found for eggs from the SS-maintained mosquitoes of both species (Ae. aegypti OR = 0.40, 95% CI 0.26-0.62; An. stephensi OR = 0.59, 95% CI 0.36-0.96). Oocyst counts were similar for P. gallinaceum infections of Ae. aegypti mosquitoes maintained on SS or BB (mean ratio = [mean on SS]/[mean on BB] = 1.11, 95% CI 0.85-1.49). Similar oocyst counts were also observed from the P. falciparum infections of SS- or BB-maintained An. stephensi (mean ratio = 0.76, 95% CI 0.44-1.37). The average counts of sporozoites/mosquito showed no evidence of reductions in the SS-maintained relative to BB-maintained mosquitoes of both species. CONCLUSIONS: Aedes aegypti and An. stephensi can be reliably maintained on SS over multiple generations and are as competent for Plasmodium infection as mosquitoes maintained on BB. Use of SS alleviates the need to acquire and preserve blood for mosquito husbandry and may support new initiatives in fundamental and applied research, including novel manipulations of midgut microbiota and factors important to the mosquito life cycle and pathogen susceptibility.

Effects of a blood-free mosquito diet on fitness and gonotrophic cycle parameters of laboratory reared Anopheles gambiae sensu stricto.

BACKGROUND: The current rise of new innovative tools for mosquito control, such as the release of transgenic mosquitoes carrying a dominant lethal gene and Wolbachia-based strategies, necessitates a massive production of mosquitoes in the insectary. However, currently laboratory rearing depends on vertebrate blood for egg production and maintenance. This practice raises ethical concerns, incurs logistical and cost limitations, and entails potential risk associated with pathogen transmission and blood storage. Consequently, an artificial blood-free diet emerges as a desirable alternative to address these challenges. This study aims to evaluate the effects of a previously formulated artificial blood-free diet (herein referred to as BLOODless) on Anopheles gambiae (An. gambiae s.s.; IFAKARA) gonotrophic parameters and fitness compared with bovine blood. METHODS: The study was a laboratory-based comparative evaluation of the fitness, fecundity and fertility of An. gambiae s.s. (IFAKARA) reared on BLOODless versus vertebrate blood from founder generation (F0) to eighth generation (F8). A total of 1000 female mosquitoes were randomly selected from F0, of which 500 mosquitoes were fed with bovine blood (control group) and the other 500 mosquitoes were fed with BLOODless diet (experimental group). The feeding success, number of eggs per female, hatching rate and pupation rate were examined post-feeding. Longevity and wing length were determined as fitness parameters for adult male and female mosquitoes for both populations. RESULTS: While blood-fed and BLOODless-fed mosquitoes showed similar feeding success, 92.3% [95% confidence interval (CI) 89.7-94.9] versus 93.6% (95% CI 90.6-96.6), respectively, significant differences emerged in their reproductive parameters. The mean number of eggs laid per female was significantly higher for blood-fed mosquitoes (P < 0.001) whereas BLOODless-fed mosquitoes had significantly lower hatching rates [odds ratio (OR) 0.17, 95% CI 0.14-0.22, P < 0.001]. Wing length and longevity were similar between both groups. CONCLUSIONS: This study demonstrates the potential of the BLOODless diet as a viable and ethical alternative to vertebrate blood feeding for rearing An. gambiae s.s. This breakthrough paves the way for more efficient and ethical studies aimed at combating malaria and other mosquito-borne diseases.

Effects of blood and root-dentin cleaning on the porosity and bond strength of a collagen bioceramic material.

To assess the effect of cleaning protocols on dentin contaminated with blood in reparative endodontic materials, bovine root samples were divided: no contamination (N); contamination (P); contamination and cleaning with saline (S), 2.5% NaOCl+saline (Na) or 2.5% NaOCl+17% EDTA+saline (NaE) and filled with: mineral trioxide aggregate (MTA), calcium-aluminate-cement (C), or C+collagen (Ccol) (n=13). The samples were evaluated for porosity, chemical composition, and bond strength. MTA porosity was lower than C (p=0.02) and higher than Ccol (p<0.001). P and NaE were similar (p=1.00), but higher than the other groups (p<0.001). MTA bond strength was similar to Ccol (p=0.777) and lower than C (p=0.028). P presented lower bond strength than the N (p<0.001); S and Na were similar to each other (p=0.969), but higher than P and lower than N (p<0.001). It was observed a predominance of mixed and cohesive failures. None of the samples showed Ca/P ratio values similar to human hydroxyapatite. This study showed that contamination with blood increased the materials porosity, but dentin cleaning with 2.5% NaOCl reduced this effect, and the collagen additive reduced the material porosity. Furthermore, blood contamination reduced the materials bond strength, and cleaning with saline or 2.5% NaOCl diminished this effect.

Suministro de sangre para transfusiones en los países de América Latina y el Caribe 2018-2020

La provisión y el acceso a sangre segura para transfusiones están en relación estrecha con la organización y el grado de desarrollo de los servicios de sangre, con la gobernanza y con la participación de la sociedad a través de la donación voluntaria no remunerada. Sin embargo, un aspecto importante al abordar la disponibilidad de sangre es la actitud solidaria de las personas que donan sangre y componentes sanguíneos de manera voluntaria y regular. A pesar del notable aumento de la donación voluntaria de sangre en la región, la donación voluntaria aún se encuentra por debajo del 50% y la disponibilidad de sangre por cada mil habitantes en algunos países está muy por debajo de la demanda estimada. Desde el 2004, la Organización Panamericana de la Salud (OPS) ha recopilado y publicado los indicadores relacionados con el suministro de sangre en los países de América Latina y el Caribe, con el fin de facilitar el seguimiento de los avances en el suministro de sangre en los países de la Región. En el 2014, los países de la Región de las Américas reafirmaron su compromiso con la salud universal a través de la aprobación del Plan de acción para el acceso universal a sangre segura 2014-2019, aprobado por el 53.o Consejo Directivo celebrado en octubre del 2014 (CD53.6). Este plan promovía el acceso universal a sangre segura para transfusiones en la Región a través de donaciones voluntarias no remuneradas, la organización de servicios de sangre y la aplicación de estándares de calidad y seguridad y de acciones de gobernanza. El informe final del plan de acción para el acceso universal a sangre segura evidenció modestos avances obtenidos en su implementación, destacándose la cobertura del 100% de análisis de la sangre que se va a transfundir, lo cual llevó a una reducción muy importante en la posibilidad de transmisión por transfusión del VIH y otras infecciones. Sin embargo, este avance puede haber disminuido la prioridad del tema de la sangre en la agenda de salud pública, dejando rezagadas las demás acciones propuestas en el plan para aumentar la seguridad transfusional. También se demostró que es necesario intensificar la integración del tema de la sangre en programas prioritarios de salud pública a fin de destacar su relevancia en temas como la mortalidad materna, los trasplantes y el control de eventos infecciosos como los relacionados con las hepatitis B y C. Así mismo, se requiere fortalecer las acciones de gobernanza en la vigilancia y la organización eficiente de los servicios de sangre para disminuir la existencia de modelos de servicios dispersos, poco eficientes y con altos costos económicos que contribuyen a mantener bajos niveles de acceso y disponibilidad de sangre y escaso avance en la donación voluntaria no remunerada, entre otras acciones necesarias para la seguridad de la sangre. Los datos presentados en esta publicación permiten monitorizar e informar con indicadores específicos el progreso y las limitaciones en la aplicación del Plan de acción para el acceso universal a sangre segura. Asimismo, se espera que estos datos promuevan el análisis y la evaluación a nivel nacional, subregional, y la toma de decisiones que fortalezcan o modifiquen las estrategias que mejoren la seguridad de la sangre y la accesibilidad a las transfusiones. La información fue proporcionada por las autoridades de los países y corresponde a los años 2018, 2019 y 2020.

Temporal dynamics of the multi-omic response to endurance exercise training.

Regular exercise promotes whole-body health and prevents disease, but the underlying molecular mechanisms are incompletely understood1-3. Here, the Molecular Transducers of Physical Activity Consortium4 profiled the temporal transcriptome, proteome, metabolome, lipidome, phosphoproteome, acetylproteome, ubiquitylproteome, epigenome and immunome in whole blood, plasma and 18 solid tissues in male and female Rattus norvegicus over eight weeks of endurance exercise training. The resulting data compendium encompasses 9,466 assays across 19 tissues, 25 molecular platforms and 4 training time points. Thousands of shared and tissue-specific molecular alterations were identified, with sex differences found in multiple tissues. Temporal multi-omic and multi-tissue analyses revealed expansive biological insights into the adaptive responses to endurance training, including widespread regulation of immune, metabolic, stress response and mitochondrial pathways. Many changes were relevant to human health, including non-alcoholic fatty liver disease, inflammatory bowel disease, cardiovascular health and tissue injury and recovery. The data and analyses presented in this study will serve as valuable resources for understanding and exploring the multi-tissue molecular effects of endurance training and are provided in a public repository ( https://motrpac-data.org/ ).

Shedding light on the effects of blood on meniscus tissue: the role of mononuclear leukocytes in mediating meniscus catabolism.

OBJECTIVE: Traumatic meniscal injuries can cause acute pain, hemarthrosis (bleeding into the joint), joint immobility, and post-traumatic osteoarthritis (PTOA). However, the exact mechanism(s) by which PTOA develops following meniscal injuries is unknown. Since meniscus tears commonly coincide with hemarthrosis, investigating the direct effects of blood and its constituents on meniscus tissue is warranted. The goal of this study was to determine the direct effects of blood and blood components on meniscus tissue catabolism. METHODS: Porcine meniscus explants or primary meniscus cells were exposed to whole blood or various fractions of blood for 3 days to simulate blood exposure following injury. Explants were then washed and cultured for an additional 3 days prior to collection for biochemical analyses. RESULTS: Whole blood increased matrix metalloproteinase (MMP) activity. Fractionation experiments revealed blood-derived red blood cells did not affect meniscus catabolism. Conversely, viable mononuclear leukocytes induced MMP activity, nitric oxide (NO) production, and loss of tissue sulfated glycosaminoglycan (sGAG) content, suggesting that these cells are mediating meniscus catabolism. CONCLUSIONS: These findings highlight the potential challenges of meniscus healing in the presence of hemarthrosis and the need for further research to elucidate the in vivo effects of blood and blood-derived mononuclear leukocytes due to both hemarthrosis and blood-derived therapeutics.

Assessment of optimal blood exposure time at 37 °C during in vitro tsetse flies feeding for quality production in mass-rearing colonies.

Control of African animal trypanosomosis is implemented through an integrated control strategy, with the sterile insect technique (SIT) as one of its components. The SIT requires mass rearing of tsetse fly colonies using an in vitro feeding system. The exposure of blood at 37 °C on heating plates over time can have an impact on the quality of fly productivity. In this study, we investigated the survival and fecundity of adult tsetse flies fed at 37 °C on 8 blood exposure times ranging from 30 min to 4 h with increments of 30 min (treatment 1, flies were fed 30 min after exposure to blood at 37 °C; treatment 2, 1 h and so on until treatment 8 [4 h after]) in order to determine the optimal exposure time. In addition, bacterial growth in blood from each treatment was assessed by agar culture at 37 °C for 72 h. The results showed that the adult female survival rates were similar regardless of the treatment. For males, only those of treatment 1 (30 min) showed a marginal lower survival than those of treatments 7 and 8 fed after 3 h 30 min and 4 h of blood exposure, respectively. Over the 4-h interval of blood exposure at 37 °C, the results showed that the number of pupae produced per initial female and pupal weight tended to increase with exposure time, but the differences were not significant. We discuss the implications of these results on tsetse mass rearing for the SIT program.

Candida auris from the Egyptian cobra: Role of snakes as potential reservoirs.

Candida auris represents one of the most urgent threats to public health, although its ecology remains largely unknown. Because amphibians and reptiles may present favorable conditions for C. auris colonization, cloacal and blood samples (n = 68), from several snake species, were cultured and molecularly screened for C. auris using molecular amplification of glycosylphosphatidylinositol protein-encoding genes and ribosomal internal transcribed spacer sequencing. Candida auris was isolated from the cloacal swab of one Egyptian cobra (Naja haje legionis) and molecularly identified in its cloaca and blood. The isolation of C. auris from wild animals is herein reported for the first time, thus suggesting the role that these animals could play as reservoirs of this emerging pathogen. The occurrence of C. auris in blood requires further investigation, although the presence of cationic antimicrobial peptides in the plasma of reptiles could play a role in reducing the vitality of the fungus.

Candida auris represents one of the most urgent threats to public health. In this study, we reported for the first time the isolation of C. auris from snake thus suggesting the role of these animals as reservoirs of this emerging pathogen.

Determinants of bacterial survival and proliferation in blood.

Bloodstream infection is a major public health concern associated with high mortality and high healthcare costs worldwide. Bacteremia can trigger fatal sepsis whose prevention, diagnosis, and management have been recognized as a global health priority by the World Health Organization. Additionally, infection control is increasingly threatened by antimicrobial resistance, which is the focus of global action plans in the framework of a One Health response. In-depth knowledge of the infection process is needed to develop efficient preventive and therapeutic measures. The pathogenesis of bloodstream infection is a dynamic process resulting from the invasion of the vascular system by bacteria, which finely regulate their metabolic pathways and virulence factors to overcome the blood immune defenses and proliferate. In this review, we highlight our current understanding of determinants of bacterial survival and proliferation in the bloodstream and discuss their interactions with the molecular and cellular components of blood.

Impact of anti-coagulant choice on blood elongational behavior.

Blood is a highly complex fluid with rheological properties that have a significant impact on various flow phenomena. In particular, it exhibits a non-Newtonian elongational viscosity that is comparable to polymer solutions. In this study, we investigate the effect of three different anticoagulants, namely EDTA (ethylene diamine tetraacetic acid), heparin, and citrate, on the elongational properties of both human and swine blood. We observe a unique two stage thinning process and a strong dependency of the characteristic relaxation time on the chosen anticoagulant, with the longest relaxation time and thus the highest elongational viscosity being found for the case of citrate. Our findings for the latter are consistent with the physiological values obtained from a dripping droplet of human blood without any anticoagulant. Furthermore, our study resolves the discrepancy found in the literature regarding the reported range of characteristic relaxation times, confirming that the elongational viscosity must be taken into account for a full rheological characterization of blood. These results have important implications for understanding blood flow in various physiological, pathological and technological conditions.

Detection of an emerging pathogen: A real time qualitative pcr assay targeting Haematospirillum jordaniae for EDTA whole blood and plasma clinical specimens.

Haematospirillum jordaniae is a gram-negative bacterium that has been identified in the blood of septic patients. The environmental source or potential zoonotic host of this bacterium, recently described as a human bacterial pathogen is unknown. An increasing number of H. jordaniae clinical infections identified by our laboratory suggested the need for an assay to detect this organism in order to aid clinical teams and practitioners with faster identification and treatment thus improving patient prognosis. Described here is a real-time qualitative PCR assay designed using gene targets identified from the analysis of 14 H. jordaniae genomes sequenced by the Center for Disease Control and Prevention's (CDC) Special Bacterial Reference Laboratory (SBRL) culture collection. The assay was validated on clinical EDTA whole blood samples as well as on plasma and determined to be effective at detecting as few as 10 copies per microliter (10,000 copies per mL, 4 log/mL) for whole blood samples and 1 copy per microliter (1,000 copies per mL, 3 log mL) for plasma samples.

An elderly male with septic arthritis and bacteremia.

Rhodococcus hoagii is a gram positive actinomycete found in horses and cattle. Humans can be infected by ingestion or inhalation through contaminated food or soil. The organism usually infects immunosuppressed hosts with pneumonia being the common presentation. We present a case of an 89 years old, apparently immunocompetent host presenting with fever, encephalopathy and arthritis who grew Rhodococcus hoagii in blood and synovial fluid, The patient responded well to a combination of vancomycin, azithromycin and imipenem-cilastatin. Our case demonstrates that extra-pulmonary manifestations such as septic arthritis and bacteremia can be seen in immune competent hosts.

The Presence of Blood in a Strain Gauge Pressure Transducer Has a Clinical Effect on the Accuracy of Intracranial Pressure Readings.

IMPORTANCE: Patients admitted with cerebral hemorrhage or cerebral edema often undergo external ventricular drain (EVD) placement to monitor and manage intracranial pressure (ICP). A strain gauge transducer accompanies the EVD to convert a pressure signal to an electrical waveform and assign a numeric value to the ICP. OBJECTIVES: This study explored ICP accuracy in the presence of blood and other viscous fluid contaminates in the transducer. DESIGN: Preclinical comparative design study. SETTING: Laboratory setting using two Natus EVDs, two strain gauge transducers, and a sealed pressure chamber. PARTICIPANTS: No human subjects or animal models were used. INTERVENTIONS: A control transducer primed with saline was compared with an investigational transducer primed with blood or with saline/glycerol mixtures in mass:mass ratios of 25%, 50%, 75%, and 100% glycerol. Volume in a sealed chamber was manipulated to reflect changes in ICP to explore the impact of contaminates on pressure measurement. MEASUREMENTS AND MAIN RESULTS: From 90 paired observations, ICP readings were statistically significantly different between the control (saline) and experimental (glycerol or blood) transducers. The time to a stable pressure reading was significantly different for saline vs. 25% glycerol (< 0.0005), 50% glycerol (< 0.005), 75% glycerol (< 0.0001), 100% glycerol (< 0.0005), and blood (< 0.0005). A difference in resting stable pressure was observed for saline vs. blood primed transducers (0.041). CONCLUSIONS AND RELEVANCE: There are statistically significant and clinically relevant differences in time to a stable pressure reading when contaminates are introduced into a closed drainage system. Changing a transducer based on the presence of blood contaminate should be considered to improve accuracy but must be weighed against the risk of introducing infection.

Unresolved haemosporidia of the Australian skink, Egernia stokesii.

The Australian skink Egernia stokesii had been recognised as a host of two species of Plasmodium, Plasmodium mackerrasae and P. circularis; nevertheless, molecular data are available for only a single haemosporidian species of this host. Its sequences are labelled as "Plasmodium sp." or "Plasmodium mackerrasae", but morphological characteristics of this isolate are unavailable. Phylogenetic analyses of these sequences placed them into the clade of the genus Haemocystidium. In this study, blood samples of six E. stokesii were analysed by both, molecular and microscopic methods to clarify the haemosporidia of this lizard. Application of these approaches offered discordant results. Whereas sequence analysis clustered our isolates with lizard species of Haemocystidium, morphology of blood stages is more akin to Plasmodium than Haemocystidium. However, limited sampling, indistinguishable nuclei/merozoites and risk of possible hidden presence of mixed infection prevent reliable species identification of detected parasites or their description as new species of Haemocystidium.

Evaluation of the feasibility for replacing sheep blood with human blood in culture media used in microbiological diagnostics.

The present study aimed to suggest the replacement of animal blood with human blood in culture media, involving alternative methods and ethical considerations, such as animal welfare, in addition to potential laboratory cost reduction. Characteristics of growth and hemolysis development were compared in different culture media, using both sheep blood and human blood. Blood types from the ABO blood group system were tested, and commercially acquired sheep blood agar was used for comparison. Bacteria of the genus Streptococcus spp., Staphylococcus aureus, Enterococcus faecalis, and Escherichia coli were tested. It was observed that growth in media with type A and O positive blood showed closer similarities to those performed in agar with sheep blood. Depending on the bacterial species, the results were either more positive or not, with faster-growing and less demanding bacteria showing better results than, for example, S. pneumoniae, which demonstrated difficulty in the growth process and hemolysis generation in human blood agar. The research suggests that in some situations, sheep blood could be replaced, especially when the goal is growth and isolation, but may not be as suitable when the objective is to analyze hemolysis or when the studied species is demanding.