Assessment of primers designed for the subspecies-specific discrimination among Babesia canis canis, Babesia canis vogeli and Babesia canis rossi by PCR assay

Canine babesiosis is an infectious disease caused by either Babesia gibsoni or Babesia canis protozoans. The latter is also classified under three different phylogenetic groups, referred to as subspecies B. canis canis, B. canis vogeli and B. canis rossi. The objective of the present study was to validate and standardize a PCR assay to discriminate the organisms at the subspecies level. First, the reference sequences of the 18S rRNA, 5.8S rRNA and 28S rRNA genes, including the internal transcribed spacer 1 (ITS1) and 2 (ITS2) of the most common species and subspecies of the genus Babesia were retrieved from the GenBank database. Subspecies-specific primers (BAB3, BAB4 and BAB5) and one genus-specific primer were designed from the alignment of the sequences. The PCR assays were evaluated in three different combinations of primer pairs in order to assure complete specificity for each reaction. The results of the tests had demonstrated effectiveness of the novel primer pairs BAB1/BAB3, BAB1/BAB4 and BAB1/BAB5 for the amplification of the subspecies-specific target fragments of 746 bp (B. c. canis), 546 bp (B. c. vogeli) and 342 bp (B. c. rossi) by PCR. The original enzymatic amplification assays with novel primers reported in this paper were confirmed to be a reliable tool for the specific discrimination among B. canis subspecies by single-step PCR assays.     

Tributaries as biodiversity preserves: An ichthyoplankton perspective from the severely impounded Upper Paraná River

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Gene pool and connectivity patterns of Pinna nobilis in the Balearic Islands (Spain,Western Mediterranean Sea): Implications for its conservation through restocking

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Use of Hydrodynamic Cavitation for Algae Removal: Effect on the Inactivation of Microalgae Belonging to Genus <Emphasis Type="Italic">Scenedesmus</Emphasis>

Excessive algae growth has generated conflicts on the use of water supplies; therefore, the focus on new technologies to remove algae from water bodies is demanding. The aim of the present study was to assess the effect of hydrodynamic cavitation on the inactivation of microalgae belonging to genus Scenedesmus. A laboratory-scale experimental apparatus was built in order to accomplish this goal; it consisted of a Venturi device designed to generate the cavitation phenomenon. Suspended microalgae samples were treated for 60 minutes under different cavitation intensities (cavitation number—Cv—ranging from 0.17 to 0.27). Results evidenced that microalgae decay over time can be modeled through first-order kinetics. The maximum removal efficiency (85%) was recorded at the highest cavitation intensity (Cv?=?0.17). The removal efficiency decreased as the cavitation number increased. Hydrodynamic cavitation was effective in inactivating Scenedesmus; it produced irreversible damages to cell morphology such as flotation spines removal, cell wall lesions, cytoplasm extravasation, and cavity formation. Assumingly, hydrodynamic cavitation has great potential to treat eutrophic water bodies. Furthermore, it represents a sustainable removal technique, since it does not produce secondary pollution.     

Effects of salinity on sexual maturity and reproduction of Poecilia velifera

Poecilia velifera is a popular fish in freshwater aquarism that naturally inhabits aquatic habitats with a wide range of salinities. Nonetheless, the effects of different salinities on the reproductive success and sex ratio of the species remain unknown. Male sex characters, sex ratio and reproductive success of P. velifera were evaluated by maintaining 160 juveniles (0.078 ± 0.011 g) in four different salinities (0; 12; 24; 36 g/L), with four replicates. The only modification observed in males was the formation of a copulatory organ from the anal fin, which was used to distinguish them from females. Timing of the formation of the male copulatory organ, and the weight and total length of males when it occurred, were recorded. Twenty‐eight fish from each treatment were euthanized after 150 day to examine the gonads and confirm sex. Eight females and four males from each treatment were then kept in their respective salinity treatments for 65 days to examine the effects of salinity on reproduction. Males were kept with females in the different treatments for 15 days, at which point they were removed. Salinity was negatively correlated with male weight and length. Salinity also affected the sex ratio, with the percentages of males and females being 32.5 and 67.5%, and 27.5 and 72.5% in the salinities of 24 and 36 g of salt/L, respectively. Fifty percent of the females kept in freshwater reproduced, while there was no reproduction in the other treatments. The maintenance of P. velifera in freshwater promoted greater reproductive success and precocity, as well as larger males.     

Lunasin and Bowman-Birk protease inhibitor concentrations of protein extracts from enzyme-assisted aqueous extraction of soybeans

Lunasin and Bowman-Birk protease inhibitor (BBI) are two soybean peptides to which health-promoting properties have been attributed. Concentrations of these peptides were determined in skim fractions produced by enzyme-assisted aqueous extraction processing (EAEP) of extruded full-fat soybean flakes (an alternative to extracting oil from soybeans with hexane) and compared with similar extracts from hexane-defatted soybean meal. Oil and protein were extracted by using countercurrent two-stage EAEP of soybeans at 1:6 solids-to-liquid ratio, 50 °C, pH 9.0, and 120 rpm for 1 h. Protein-rich skim fractions were produced from extruded full-fat soybean flakes using different enzyme strategies in EAEP: 0.5% protease (wt/g extruded flakes) used in both extraction stages; 0.5% protease used only in the second extraction stage; no enzyme used in either extraction stage. Countercurrent two-stage protein extraction of air-desolventized, hexane-defatted soybean flakes was used as a control. Protein extraction yields increased from 66% to 89-96% when using countercurrent two-stage EAEP with extruded full-fat flakes compared to 85% when using countercurrent two-stage protein extraction of air-desolventized, hexane-defatted soybean flakes. Extruding full-fat soybean flakes reduced BBI activity. Enzymatic hydrolysis reduced BBI contents of EAEP skims. Lunasin, however, was more resistant to both enzymatic hydrolysis and heat denaturation. Although using enzymes in both EAEP extraction stages yielded the highest protein and oil extractions, reducing enzyme use to only the second stage preserved much of the BBI and Lunasin.     

Acute bacterial cystitis does not cause deoxyribonucleic acid damage detectable by the alkaline comet assay in urothelial cells of dogs

Considering the high incidence of dogs with acute bacterial cystitis (BC) and the relationship among inflammation, genotoxicity, and carcinogenesis, we conducted a case-control study comparing the frequency of deoxyribonucleic acid (DNA) lesions assessed by the comet assay between disease-free animals (13 males and 13 females) and cytology-confirmed cases of acute BC (12 males and 12 females), which was mainly caused by Staphylococcus sp. (40%) and Escherichia coli (35%). The results show no increase in DNA damage in cells obtained by bladder washings and no influence of age, sex, and breed due to acute BC. In conclusion, DNA damage was seemingly not associated with the infection by specific bacteria.