Microbial Flora and Nutrient Content of Market Bought Smoked African Cat Fish Clarias gariepinus from Jos, Nigeria

Clarias gariepinus, one of the many fishes sold in Nigerian market, is the most preferred smoked fish in Jos where large quantities are smoked and stored for sale. This study assessed the nutritional value and health of smoked C. gariepinus sold in Jos markets.Live and smoked C. gariepinus were purchased from the four major markets in Jos metropolis. Microorganisms isolated from the smoked fish were identified. The live fish were smoked in the laboratory and inoculated with the isolated microorganisms. Nutrient content of the fishes were monitored weekly for four weeks, un-inoculated laboratory smoked fish served as controls.Bacillus brevis, Aspergillus fumigates and Mucor species were isolated from purchased smoked fish. The nutrient value of these fish were significantly lower (p<0.05) than the laboratory smoked fish. There was however a gradual decline in the nutrient content of the infected laboratory smoked fish. The carbohydrate content decreased to zero while the moisture content increased. Mucor had the most significant effect on protein (62.06 ±13.39) and carbohydrate (1.11±0.95) levels in infected fish. pH dropped below7.0 by the end of four weeks in Mucor infected fish and fat content was lowest (14.19±3.82) in A. fumigates infected fish. There was a significant difference (p>0.05) between the nutrient values in the control and infected fish. The microbial content and lower nutrient values of infected fish emphasize the need to ascertain the health and nutrient content of market sold fish. This will ensure that consumers receive optimum nourishment and avoid the likely health implications of consuming infected fish. Keywords: Fish nutrient, smoked Clarias gariepinus, microflora, Jos market

Extracellular release of acid phosphatase from blood stream forms of Trypanosoma brucei brucei .

Acid phosphatase (ACP) activity was demonstrated in blood stream form of Trypanosoma brucei brucei harvested from infected Wister rats by Ion Exchange DEAE Cellulose 52 chromatography. Whole parasite extract (WPE) and Excretory Secretory Extract (ESE) were prepared and analyzed for acid phosphatase activity. A higher ACP activity (85.5 \u3bcmol/min) was recorded in WPE compared to ESE (36.8 \u3bcmol/min). ACP activity in ESE is suggestive of the presence of a cell rich enzyme. Phase separation of the extracts using the detergent Triton X-114 (TX-114), resulted in protein partitioning into aqueous and detergent phases. ACP activity was higher in the detergent phases (56.2 \u3bcmol/min and 28.8 \u3bcmol/min) of WPE and ESE respectively. ACP activity recorded in the aqueous phases of WPE and EPE was 27.8 and 7.6 \u3bcmol/min respectively. On a Size Exclusion chromatography column using Sephacryl-300, ESE emerged as five distinct protein peaks. ACP activity of the eluted fractions showed two peaks of relative molecular weights 195 and 325 KD. This study shows that T. brucei releases acid phosphatase extracellularly via a yet to be determined mechanism. Acid phosphatase activity in ESE is indicative of a soluble enzyme within the cell matrix which may also play an important role in the pathology of African Trypanosomiasis

Assessment of heavy metal residues in water, fish tissue and human blood from Ubeji, Warri, Delta State, Nigeria

Residual levels of lead, chromium, cadmium and zinc in water and fish tissue from Ubeji River, Warri and blood samples from residents of Ubeji were analysed. Control water and fish samples were obtained from Eleyele River and blood from residents of Ibadan. All the samples collected were digested using a modified procedure from the Association of Official Analytical Chemists (AOAC) and were subsequently analyzed using Atomic Absorption Spectroscopy. Levels of lead and cadmium in Ubeji river were significantly higher than levels in Eleyele River. Fifty fish samples from five species ( Citharinus citharus , Liza falcipinis , Brycinus macrolepidotus , Polydactylus quadrifilis and Tilapia zilli ) were analysed. Lead concentration (ppm) was highest in C. citharus (76.07\ub1161.48) and least in T. zilli (23.16\ub126.30). Chromium was not detected in B. macrolepidotus, P. quadrifilis and T. zilli. Concentrations of all heavy metals detected, except zinc, in the fish species were higher than WHO permissible limit. Lead, Cadmium and Zinc concentrations in the tissues of the control fish were significantly lower (p>0.05) than those recorded in fish from Ubeji River. There were no statistically significant differences in heavy metal concentration in fish gills and muscle (p>0.05). Lead and cadmium levels in Ubeji river significantly exceeded (p=0.0 and p=0.012) the permissible limits for aquaculture and drinking. Levels of Chromium (1737.17\ub12996.01), Zinc and Lead (149.35\ub1188.28 and 123.49\ub1350.85 respectively) were significantly high (p<0.05) in the 101 human blood samples screened, while concentrations of Cadmium was comparatively low (10.11\ub110.71). Lower levels of cadmium, zinc and lead (8.54\ub1 7.49, 79.89\ub162.65 and 53.46\ub157.17, respectively) were recorded for the control samples from Ibadan. This study highlights the high levels of heavy metals in the Ubeji River and suggests the need for interventions to stem the tide of pollution in the river. It would also be important to assess health problems that may arise as a result of contact and continuous use of the water. @ JASE

Comparison of PCR-based detection of Plasmodium falciparum infections based on single and multicopy genes

PCR-based assays are the most sensitive and specific methods to detect malaria parasites

Cellular responses to modified Plasmodium falciparum MSP119 antigens in individuals previously exposed to natural malaria infection

<p>Abstract</p> <p>Background</p> <p>MSP1 processing-inhibitory antibodies bind to epitopes on the 19 kDa C-terminal region of the <it>Plasmodium falciparum </it>merozoite surface protein 1 (MSP1₁₉), inhibiting erythrocyte invasion. Blocking antibodies also bind to this antigen but prevent inhibitory antibodies binding, allowing invasion to proceed. Recombinant MSP1₁₉had been modified previously to allow inhibitory but not blocking antibodies to continue to bind. Immunization with these modified proteins, therefore, has the potential to induce more effective protective antibodies. However, it was unclear whether the modification of MSP1₁₉would affect critical T-cell responses to epitopes in this antigen.</p> <p>Methods</p> <p>The cellular responses to wild-type MSP1₁₉and a panel of modified MSP1₁₉antigens were measured using an <it>in-vitro </it>assay for two groups of individuals: the first were malaria-naïve and the second had been naturally exposed to <it>Plasmodium falciparum </it>infection. The cellular responses to the modified proteins were examined using cells from malaria-exposed infants and adults.</p> <p>Results</p> <p>Interestingly, stimulation indices (SI) for responses induced by some of the modified proteins were at least two-fold higher than those elicited by the wild-type MSP1₁₉. A protein with four amino acid substitutions (Glu27→Tyr, Leu31→Arg, Tyr34→Ser and Glu43→Leu) had the highest stimulation index (SI up to 360) and induced large responses in 64% of the samples that had significant cellular responses to the modified proteins.</p> <p>Conclusion</p> <p>This study suggests that specific MSP1₁₉variants that have been engineered to improve their antigenicity for inhibitory antibodies, retain T-cell epitopes and the ability to induce cellular responses. These proteins are candidates for the development of MSP1-based malaria vaccines.</p

Assessment of heavy metal residues in water, fish tissue and human blood from Ubeji, Warri, Delta State, Nigeria

Residual levels of lead, chromium, cadmium and zinc in water and fish tissue from Ubeji River, Warri and blood samples from residents of Ubeji were analysed. Control water and fish samples were obtained from Eleyele River and blood from residents of Ibadan. All the samples collected were digested using a modified procedure from the Association of Official Analytical Chemists (AOAC) and were subsequently analyzed using Atomic Absorption Spectroscopy. Levels of lead and cadmium in Ubeji river were significantly higher than levels in Eleyele River. Fifty fish samples from five species ( Citharinus citharus , Liza falcipinis , Brycinus macrolepidotus , Polydactylus quadrifilis and Tilapia zilli ) were analysed. Lead concentration (ppm) was highest in C. citharus (76.07±161.48) and least in T. zilli (23.16±26.30). Chromium was not detected in B. macrolepidotus, P. quadrifilis and T. zilli. Concentrations of all heavy metals detected, except zinc, in the fish species were higher than WHO permissible limit. Lead, Cadmium and Zinc concentrations in the tissues of the control fish were significantly lower (p>0.05) than those recorded in fish from Ubeji River. There were no statistically significant differences in heavy metal concentration in fish gills and muscle (p>0.05). Lead and cadmium levels in Ubeji river significantly exceeded (p=0.0 and p=0.012) the permissible limits for aquaculture and drinking. Levels of Chromium (1737.17±2996.01), Zinc and Lead (149.35±188.28 and 123.49±350.85 respectively) were significantly high (p<0.05) in the 101 human blood samples screened, while concentrations of Cadmium was comparatively low (10.11±10.71). Lower levels of cadmium, zinc and lead (8.54± 7.49, 79.89±62.65 and 53.46±57.17, respectively) were recorded for the control samples from Ibadan. This study highlights the high levels of heavy metals in the Ubeji River and suggests the need for interventions to stem the tide of pollution in the river. It would also be important to assess health problems that may arise as a result of contact and continuous use of the water. @ JASE

Extracellular release of acid phosphatase from blood stream forms of Trypanosoma brucei brucei .

Acid phosphatase (ACP) activity was demonstrated in blood stream form of Trypanosoma brucei brucei harvested from infected Wister rats by Ion Exchange DEAE Cellulose 52 chromatography. Whole parasite extract (WPE) and Excretory Secretory Extract (ESE) were prepared and analyzed for acid phosphatase activity. A higher ACP activity (85.5 μmol/min) was recorded in WPE compared to ESE (36.8 μmol/min). ACP activity in ESE is suggestive of the presence of a cell rich enzyme. Phase separation of the extracts using the detergent Triton X-114 (TX-114), resulted in protein partitioning into aqueous and detergent phases. ACP activity was higher in the detergent phases (56.2 μmol/min and 28.8 μmol/min) of WPE and ESE respectively. ACP activity recorded in the aqueous phases of WPE and EPE was 27.8 and 7.6 μmol/min respectively. On a Size Exclusion chromatography column using Sephacryl-300, ESE emerged as five distinct protein peaks. ACP activity of the eluted fractions showed two peaks of relative molecular weights 195 and 325 KD. This study shows that T. brucei releases acid phosphatase extracellularly via a yet to be determined mechanism. Acid phosphatase activity in ESE is indicative of a soluble enzyme within the cell matrix which may also play an important role in the pathology of African Trypanosomiasis

Assessment of heavy metal residues in water, fish tissue and human blood from Ubeji, Warri, Delta State, Nigeria

Residual levels of lead, chromium, cadmium and zinc in water and fish tissue from Ubeji River, Warri and blood samples from residents of Ubeji were analysed. Control water and fish samples were obtained from Eleyele River and blood from residents of Ibadan. All the samples collected were digested using a modified procedure from the Association of Official Analytical Chemists (AOAC) and were subsequently analyzed using Atomic Absorption Spectroscopy. Levels of lead and cadmium in Ubeji river were significantly higher than levels in Eleyele River. Fifty fish samples from five species ( Citharinus citharus , Liza falcipinis , Brycinus macrolepidotus , Polydactylus quadrifilis and Tilapia zilli ) were analysed. Lead concentration (ppm) was highest in C. citharus (76.07±161.48) and least in T. zilli (23.16±26.30). Chromium was not detected in B. macrolepidotus, P. quadrifilis and T. zilli. Concentrations of all heavy metals detected, except zinc, in the fish species were higher than WHO permissible limit. Lead, Cadmium and Zinc concentrations in the tissues of the control fish were significantly lower (p>0.05) than those recorded in fish from Ubeji River. There were no statistically significant differences in heavy metal concentration in fish gills and muscle (p>0.05). Lead and cadmium levels in Ubeji river significantly exceeded (p=0.0 and p=0.012) the permissible limits for aquaculture and drinking. Levels of Chromium (1737.17±2996.01), Zinc and Lead (149.35±188.28 and 123.49±350.85 respectively) were significantly high (p<0.05) in the 101 human blood samples screened, while concentrations of Cadmium was comparatively low (10.11±10.71). Lower levels of cadmium, zinc and lead (8.54± 7.49, 79.89±62.65 and 53.46±57.17, respectively) were recorded for the control samples from Ibadan. This study highlights the high levels of heavy metals in the Ubeji River and suggests the need for interventions to stem the tide of pollution in the river. It would also be important to assess health problems that may arise as a result of contact and continuous use of the water. @ JASE

DNA methylation profiles in urothelial bladder cancer tissues and children with schistosomiasis from Eggua, Ogun State, Nigeria

Abstract Background Squamous cell carcinoma has been attributed to chronic schistosomiasis and is the predominant type of bladder cancer in schistosomiasis endemic areas. The aim of this study was to assess early promoter DNA methylation in selected genes implicated in schistosomiasis-associated bladder cancer (SABC). Methods A total of 159 urine samples were collected from school-aged children in Eggua Community of Ogun State and examined by microscopy for Schistosoma haematobium eggs. From this sample, a subset of 34 (21.1%) urine samples positive for S. haematobium, age and sex-matched with negative urine control samples, and 16 formalin-fixed paraffin-embedded bladder cancer tissues obtained from the University College Hospital were subjected to DNA isolation and bisulphite DNA conversion. Quantitative methylation-specific PCR was used to determine the methylation status of APC, RARβ2, RASSF1A, and TIMP3 in the samples. Results High degrees of methylation of RARβ2(67.7%), RASSF1A (38.2%), and TIMP3(52.9%) was more common in urogenital schistosomiasis (UGS)-positive urine samples than negative urine (control) samples and in bladder cancer tissues. Promoter DNA methylation in the positive urine samples was 1.4-fold, 13.3-fold, 3.4-fold, and 3.8-fold higher in APC, RARβ2, RASSF1A, and TIMP3, respectively, than in the matched controls. The odds of promoter methylation were likely to increase with age group for APC (OR: 1.615) and TIMP3(OR: 2.000); sex for TIMP3(OR: 2.644); and haematuria for RARβ2(OR: 1.094), RASSF1A (OR: 1.143), and TIMP3(OR: 1.842), although there were no significant associations. Conclusions: Gene promoter DNA methylation in tumour suppressor genes was observed in schistosomiasis cases. Hence, promoter DNA methylation may occur during active schistosomiasis in children. This result may serve as an early non-invasive biomarker to detect and hint at the risk of developing SABC later in life

Comparison of PCR-based detection of <it>Plasmodium falciparum </it>infections based on single and multicopy genes

Abstract PCR-based assays are the most sensitive and specific methods to detect malaria parasites. This study compared the diagnostic accuracy of three PCR-based assays that do not only differ in their sequence target, but also in the number of copies of their target region, for the detection of Plasmodium falciparum in 401 individuals living in a malaria-endemic area in Nigeria. Compared to a composite reference generated from results of all the 3 PCR assays, the stevor gene amplification had a sensitivity of 100% (Kappa = 1; 95% CI = 1.000–1.000), 83% (Kappa = 0.718; 95% CI = 0.648–0.788) by SSUrRNA gene PCR and 71% (Kappa = 0.552; 95% CI = 0.478–0.627) by the msa-2 gene amplification. Results from this study indicate that the stevor gene amplification is the most sensitive technique for the detection of P. falciparum. This assay may be an important reference standard, especially when a confirmatory technique with high sensitivity and specificity is needed for ruling out P. falciparum infection.</p