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Summary: The application of metabarcoding to study animal‐associated microeukaryotes has been restricted because the universal barcode used to study microeukaryotic ecology and distribution in the environment, the Small Subunit of the Ribosomal RNA gene (18S rRNA), is also present in the host. As a result, when host‐associated microbial eukaryotes are analysed by metabarcoding, the reads tend to be dominated by host sequences. We have done an in silico validation against the SILVA 18S rRNA database of a non‐metazoan primer set (primers that are biased against the metazoan 18S rRNA) that recovers only 2.6% of all the metazoan sequences, while recovering most of the other eukaryotes (80.4%). Among metazoans, the non‐metazoan primers are predicted to amplify 74% of Porifera sequences, 4% of Ctenophora, and 15% of Cnidaria, while amplifying almost no sequences within Bilateria. In vivo, these non‐metazoan primers reduce significantly the animal signal from coral and human samples, and when compared against universal primers provide at worst a 2‐fold decrease in the number of metazoan reads and at best a 2800‐fold decrease. This easy, inexpensive, and near‐universal method for the study of animal‐associated microeukaryotes diversity will contribute to a better understanding of the microbiome. [ABSTRACT FROM AUTHOR]

No studies are available about biochemical characteristics and modulation (i.e. by endogenous and/or environmental cues) of trypsin (a key digestive endoprotease) in hepatopancreas of intertidal euryhaline crabs neither on the possible concomitant modulation of key ectoproteases such as aminopeptidase-N (APN) involved in final steps of protein digestion. Furthermore, nothing is still known in decapods crustaceans about the role of histamine as primary chemical messenger for modulation of main components of digestive process (i.e. proteases). We determined biochemical characteristics and investigated the effect of histamine injections; of histamine in vitro and of acclimation of individuals to low and high salinity on trypsin and aminopeptidase-N (APN) activities in the hepatopancreas of the euryhaline crab Cyrtograpsus angulatus (Dana 1851). Trypsin activity was maximal at pH 7.4 and at 45 °C. APN activity increased from pH 6.6 to 7.6–9.0 and was maintained high at 37–45 °C. Both activities exhibited Michaelis-Menten kinetics (apparent Km: trypsin = 0.36 mM; APN = 0.07 mM). The injection of 10 − 4 M histamine decreased trypsin activity (about 40%) in hepatopancreas while did not affect APN activity. Similarly, in vitro 10 -4 M histamine decreased trypsin activity (about 52%) in hepatopancreas but not APN activity. Trypsin activity in the hepatopancreas was not affected by acclimation of crabs to low (10 psu) or high (40 psu) environmental salinity while APN activity was increased (about 200%) in 10 psu. The results show the differential modulation of trypsin and APN by distinct cues and point to histamine as modulator of intracellular trypsin by direct action on the hepatopancreas. [ABSTRACT FROM AUTHOR]

Background: The lack of an effective diagnostic tool for Carrion’s disease leads to misdiagnosis, wrong treatments and perpetuation of asymptomatic carriers living in endemic areas. Conventional PCR approaches have been reported as a diagnostic technique. However, the detection limit of these techniques is not clear as well as if its usefulness in low bacteriemia cases. The aim of this study was to evaluate the detection limit of 3 PCR approaches. Methodology/Principal Findings: We determined the detection limit of 3 different PCR approaches: Bartonella-specific 16S rRNA, fla and its genes. We also evaluated the viability of dry blood spots to be used as a sample transport system. Our results show that 16S rRNA PCR is the approach with a lowest detection limit, 5 CFU/μL, and thus, the best diagnostic PCR tool studied. Dry blood spots diminish the sensitivity of the assay. Conclusions/Significance: From the tested PCRs, the 16S rRNA PCR-approach is the best to be used in the direct blood detection of acute cases of Carrion’s disease. However its use in samples from dry blood spots results in easier management of transport samples in rural areas, a slight decrease in the sensitivity was observed. The usefulness to detect by PCR the presence of low-bacteriemic or asymptomatic carriers is doubtful, showing the need to search for new more sensible techniques. [ABSTRACT FROM AUTHOR]

We studied the existence, biochemical characteristics and response to different environmental salinities of amylase, maltase and sucrase activity in the intertidal euryhaline crab Cyrtograpsus angulatus (Dana, 1852) along with the response to distinct salinities of glycogen and free glucose content in storage organs. Amylase, maltase and sucrase activities were kept over a broad range of pH and temperature and exhibited Michaelis–Menten kinetics. Zymography showed the existence of two amylase forms in crabs exposed to 35 (osmoconformation) and low (6–10 psu; hyper-regulation) or high (40 psu) (hypo-regulation) salinities. Carbohydrases activity in the hepatopancreas and glycemia were not affected in crab exposed to different environmental salinities. In 6 and 40 psu, the glycogen content in anterior gills was lower than in 35 psu. In 6, 10 and 40 psu, glycogen concentration in hepatopancreas, muscle and posterior gills were similar to that in 35 psu. Free glucose concentration in chela muscle was higher in 6 and 40 psu than in 35 psu. The existence and biochemical characteristics of carbohydrases activity and the adjustments in concentration of glycogen in anterior gills and free glucose in chela muscle suggests the ability to perform complete hydrolysis of glycogenic substrates and to keep glucose homeostasis in relation to acclimation to different salinity conditions. [ABSTRACT FROM AUTHOR]

A 22-day-old male was admitted with a 2-day history of irritability, dyspnea, jaundice, fever, and gastrointestinal bleeding. A thin blood smear was performed, which showed the presence of intraerythrocyte bacteria identified as Bartonella bacilliformis, and subsequently, the child was diagnosed with Carrion's disease. The diagnosis was confirmed by specific polymerase chain reaction. The child was born in a non-endemic B. bacilliformis area and had not traveled to such an area before hospitalization. However, the mother was from an endemic B. bacilliformis area, and posterior physical examination showed the presence of a wart compatible with B. bacilliformis in semi-immune subjects. These data support vertical transmission of B. bacilliformis., (© The American Society of Tropical Medicine and Hygiene.)