The introduction of invasive Nile tilapia (Oreochromis niloticus), and the rapacious predator Nile perch (Lates niloticus), into Lake Victoria resulted in a decline in population sizes, genetic diversity and even extirpation of native species which were previously the mainstay of local fisheries. However, remnant populations of native fish species, including tilapia, still persist in satellite lakes around Lake Victoria where they may coexist with O. niloticus. In this study we assessed population genetic structure, diversity, and integrity of the native critically endangered Singidia tilapia (O. esculentus) in its refugial populations in the Yala swamp, Kenya, and contrasted this diversity with populations of the invasive tilapia O. niloticus in satellite lakes (Kanyaboli, Namboyo and Sare) and Lake Victoria. Based on mtDNA control region sequences and eight nuclear microsatellite loci, we did not detect any mtDNA introgression between the native and the invasive species in Lakes Kanyaboli and Namboyo, but did find low levels of nuclear admixture, primarily from O. niloticus to O. esculentus. Some genetic signal of O. esculentus in O. niloticus was found in Lake Sare, where O. esculentus is not found, suggesting it has recently been extirpated by the O. niloticus invasion. In both species, populations in the satellite lakes are significantly genetically isolated from each other, with private mtDNA haplotypes and microsatellite alleles. For O. niloticus, genetic diversity in satellite lakes was similar to that found in Lake Victoria. Our data imply a low frequency of immigration exchange between the two populations of O. esculentus and we suggest that the populations of this endangered species and important fisheries resource should be conserved separately in Lakes Kanyaboli and Namboyo and with high priority.
bIn combination with antibiotics, quinine is recommended as the second-line treatment for uncomplicated malaria, an alternative first-line treatment for severe malaria, and for treatment of malaria in the first trimester of pregnancy. Quinine has been shown to have frequent clinical failures, and yet the mechanisms of action and resistance have not been fully elucidated. However, resistance is linked to polymorphisms in multiple genes, including multidrug resistance 1 (Pfmdr1), the chloroquine resistance transporter (Pfcrt), and the sodium/hydrogen exchanger gene (Pfnhe1). Here, we investigated the association between in vitro quinine susceptibility and genetic polymorphisms in Pfmdr1codons 86 and 184, Pfcrt codon 76, and Pfnhe1 ms4760 in 88 field isolates from western Kenya. In vitro activity was assessed based on the drug concentration that inhibited 50% of parasite growth (the IC 50 ), and parasite genetic polymorphisms were determined from DNA sequencing. Data revealed there were significant associations between polymorphism in Pfmdr1-86Y, Pfmdr1-184F, or Pfcrt-76T and quinine susceptibility (P < 0.0001 for all three associations). Eighty-two percent of parasites resistant to quinine carried mutant alleles at these codons (Pfmdr1-86Y, Pfmdr1-184F, and Pfcrt-76T), whereas 74% of parasites susceptible to quinine carried the wild-type allele (Pfmdr1-N86, Pfmdr1-Y184, and Pfcrt-K76, respectively). In addition, quinine IC 50 values for parasites with Pfnhe1 ms4760 3 DNNND repeats were significantly higher than for those with 1 or 2 repeats (P ؍ 0.033 and P ؍ 0.0043, respectively). Clinical efficacy studies are now required to confirm the validity of these markers and the importance of parasite genetic background.
BackgroundOver 65% of human infections are ascribed to bacterial biofilms that are often highly resistant to antibiotics and host immunity. Staphylococcus epidermidis is the predominant cause of recurrent nosocomial and biofilm-related infections. However, the susceptibility patterns of S. epidermidis biofilms to physico-chemical stress induced by commonly recommended disinfectants [(heat, sodium chloride (NaCl), sodium hypochlorite (NaOCl) and hydrogen peroxide (H2O2)] in domestic and human healthcare settings remains largely unknown. Further, the molecular mechanisms of bacterial biofilms resistance to the physico-chemical stresses remain unclear. Growing evidence demonstrates that extracellular DNA (eDNA) protects bacterial biofilms against antibiotics. However, the role of eDNA as a potential mechanism underlying S. epidermidis biofilms resistance to physico-chemical stress exposure is yet to be understood. Therefore, this study aimed to evaluate the susceptibility patterns of and eDNA release by S. epidermidis biofilm and planktonic cells to physico-chemical stress exposure.ResultsS. epidermidis biofilms exposed to physico-chemical stress conditions commonly recommended for disinfection [heat (60 °C), 1.72 M NaCl, solution containing 150 μL of waterguard (0.178 M NaOCl) in 1 L of water or 1.77 M H2O2] for 30 and 60 min exhibited lower log reductions of CFU/mL than the corresponding planktonic cells (p < 0.0001). The eDNA released by sub-lethal heat (50 °C)-treated S. epidermidis biofilm and planktonic cells was not statistically different (p = 0.8501). However, 50 °C-treated S. epidermidis biofilm cells released significantly increased eDNA than the untreated controls (p = 0.0098). The eDNA released by 0.8 M NaCl-treated S. epidermidis biofilm and planktonic cells was not significantly different (p = 0.9697). Conversely, 5 mM NaOCl-treated S. epidermidis biofilms exhibited significantly increased eDNA release than the corresponding planktonic cells (p = 0.0015). Further, the 50 μM H2O2-treated S. epidermidis biofilms released significantly more eDNA than the corresponding planktonic cells (p = 0.021).ConclusionsS. epidermidis biofilms were less susceptible to physico-chemical stress induced by the four commonly recommended disinfectants than the analogous planktonic cells. Further, S. epidermidis biofilms enhanced eDNA release in response to the sub-lethal heat and oxidative stress exposure than the corresponding planktonic cells suggesting a role of eDNA in biofilms resistance to the physico-chemical stresses.
Effects of high stocking densities (HSDs) were evaluated for Nile tilapia fish (Oreochromis niloticus) under culture to determine its influence on plasma cortisol and whole blood glucose concentration. Plasma cortisol levels (ng/ml) were assayed by Enzyme-Linked Immunosorbent Assay (ELISA). Whole blood glucose levels were determined using a hand-held one touch ultraglucose meter (MD-300) and test strips. Plasma cortisol and whole blood glucose level determinations were replicated three times for O. niloticus reared under both low stocking densities (LSD) and HSD. One way Analysis of Variance (ANOVA) was performed on the data collected, and comparison of significant differences in means was carried out between LSD and HSD at 0.01%. Plasma cortisol levels revealed statistically (P≤0.01) significant values of HSD at 6.32 ± 1.06 ng/ml than in LSD at 4.62 ± 1.58 ng/ml for the O. niloticus groups studied. Whole blood glucose analysis revealed a statistical (P<0.05) difference in the means in HSD and LSD O. niloticus groups (F(df,1; 8) = 7.946 > Fcrit = 4.414; P=0.01). Mean plasma glucose concentration was statistically (P≤0.01) higher for HSD than LSD O. niloticus groups at mean ± SD, 96.84 ± 5.28 and 76.82 ± 5.92, respectively. The findings of this study demonstrate that high stocking densities increase both cortisol and whole blood glucose concentration in tilapia fish, indicating a marked increase in stress levels. Elevated plasma cortisol and whole blood glucose concentration can be used as biomarkers for acute stress in O. niloticus produced under aquaculture systems. The findings of this study can help inform policy on the management of stress caused by overstocking of O. niloticus and other related Cichlids under industrial aquaculture production.
Fish farmers have a tendency of employing high stocking density (HSD) as a means of increasing productivity. However, HSD is a chronic stressor that is likely to lower profitability of fish farming if not implemented properly. HSD induces stress which in turn elevates sequentially the levels of plasma cortisol and glucose. The resultant glucose is distributed to various tissues by glucose transporter protein (GLUTs) to restore normalcy. GLUT 1, transmembrane protein found in erythrocytes, is responsible for import and export of glucose in red blood cells. However, knowledge on how chronic stress impact glucose and GLUT 1 protein in fish subjected to HSD is still unclear. In this study, effect of HSD on the expression of GLUT 1 in Nile tilapia was investigated in an attempt to elucidate the role of GLUT 1 in glucose metabolism during chronic stress. Fish were reared for 4 weeks at 1.5 and 4.5 kg/m 3 for low stocking density (LSD) and HSD, respectively. Four physiological parameters were determined from the blood samples obtained from fish at the end of experiment. At p \ 0.05, there were significant differences between fish reared at HSD and LSD in plasma cortisol level (72.1 ± 5.9 ng/ml and 37.5 ± 4.6 ng/ml); blood glucose level (136.00 ± 1.3 mg/dL and 70.2 ± 1.0 mg/dL); erythrocytes count (7.2 ± 0.5 9 106 mm -3 and 2.1 ± 0.4 9 106 mm -3 ); and plasma GLUT 1 level (1.40 ± 0.17 rbi and 0.81 ± 0.07 rbi), respectively. HSD induced elevation of plasma cortisol level, blood glucose level, erythrocytes count and GLUT 1 level. These elevated physiological factors and particularly GLUT 1 can be used as a cellular stress biomarker in fish farming and aquaculture.
Staphylococcus epidermidis is the predominant cause of recalcitrant biofilm-associated infections, which are often highly resistant to antibiotics. Thus, the use of physico-chemical agents for disinfection offers a more effective approach to the control of S. epidermidis biofilm infections. However, the underlying tolerance mechanisms employed by S. epidermidis biofilm against these physico-chemical disinfectants remain largely unknown. The expression of a σ B -dependent gene, alkaline shock protein 23 ( asp23 ) and catalase activity by S. epidermidis biofilm and planktonic cells exposed to heat (50 °C), 0.8 M sodium chloride (NaCl), 5 mM sodium hypochlorite (NaOCl) or 50 μM hydrogen peroxide (H 2 O 2 ) for 60 minutes were compared. Significantly higher asp23 expression levels were observed in biofilms exposed to 50 °C, 5 mM NaOCl or 50 μM H 2 O 2 compared to the corresponding planktonic cells ( p < 0.05). Conversely, asp23 expression levels in biofilm and planktonic cells exposed to 0.8 M NaCl were not significantly different ( p > 0.05). Further, biofilms exposed to 50 °C, 0.8 M NaCl, 5 mM NaOCl or 50 μM H 2 O 2 exhibited significantly higher catalase activity than the planktonic cells ( p < 0.05). These results suggest that activities of σ B and catalase may be involved in the tolerance of S. epidermidis biofilm against physico-chemical disinfection.
We carried out a postrelease evaluation to determine predictors of habitat use and carrying capacity for the black rhinoceros (Diceros bicornis michaeli), which are critical for monitoring how the Ruma National Park sub‐population may contribute to Kenya's meta‐population strategy. We determined whether level of elevation, rockiness, shade, distance to fence, roads, and human settlements predict habitat use, differences in habitat and diet preference between female and male black rhinoceros, and the ecological carrying capacity (CC) of black rhinoceros in the park. We used standard ecological methods to collect data on predictors of habitat use, habitat preference and to estimate CC. Results show, first, that none of the environmental and anthropogenic factors evaluated predicted habitat use by black rhinoceros in the park. Second, although there was no significant difference in habitat preference between the sexes (U = 16.50, p = 0.306), there was a 60% difference in Jaccard's dissimilarity in diet selection between the sexes. Third, the park can support 65 black rhinoceros. Altogether, the findings suggest that the park has potential to support other sub‐populations in Kenya. We recommend that future similar studies should incorporate population viability analysis and a community‐based approach to forecast the species health and extinction risk.
The novelty and suitability of the mitochondrial gene CO1 in DNA barcoding as a reliable identification tool in animal species are undisputed. This is attributed to its standardized sequencing segment of the mitochondrial cytochrome c oxidase-1 gene (CO1) which has the necessary universality and variability making it a generally acceptable barcode region. CO1 is a haploid single locus that is uniparentally-inherited. Protein-coding regions are present in high-copy numbers making it an ideal barcode. The mitochondrial oxidase subunit I (COI) gene is a robust barcode with a suitable threshold for delineating animals and is not subject to drastic length variation, frequent mononucleotide repeats or microinversions. However, a low nucleotide substitution rate of plant mitochondrial genome [mtDNA] precludes the use of CO1 as a universal plant DNA barcode and makes the search for alternative barcode regions necessary. Currently, there exists no universal barcode for plants. The plastid region reveals leading candidate loci as appropriate DNA barcodes yet to be explored in biodiversity studies in Kenya. Four of these plastid regions are portions of coding genes (matK, rbcL, rpoB, and rpoC1), and three noncoding spacers (atpF-atpH, trnH-psbA, and psbK-psbL) which emerge as ideal candidate DNA loci. While dif
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