Design and evaluation of primer pairs for efficient detection of avian rotavirus

The use of molecular methods for rotavirus characterisation provides increased sensitivity for typing and allows the identification of putative reassortant strains. Reagents and methods for genotyping the virus need constant modification because of the reassortant nature of the virus. This study was aimed at designing and evaluating new oligonucleotide degenerate primer pairs that provide increased sensitivity and specificity for detecting avian rotavirus. Gene-specific primer pairs were designed by analysing different rotavirus strains isolated during the last decade by downloading them from the GenBank. The alignments were generated using clustal analysis from the BioEdit program. Degenerate nucleotides were included due to the reassortant nature of rotavirus. The consensus sequences were aligned using the BioEdit program and then treated with the Fast PCR software to derive the primers. The derived primer sequences were submitted for a BLAST search to ensure alignment was exclusive to the desired target genes. The designed primers had specific bands and were efficient in detecting rotavirus in faecal samples than previously published primers. Thus, a successful surveillance of rotaviruses requires that primer pairs be updated regularly in order to detect the emergence of novel or “unusual types”, which have occurred by genetic drift causing nucleotide changes at the primer binding sites that result in typing failures. We recommend the use of the proposed primers in molecular surveillance studies for efficient detection of avian rotavirus.     

Physicochemical and kinetic characteristics of rhodanese from the liver of African catfish <Emphasis Type="Italic">Clarias gariepinus</Emphasis> Burchell in Asejire lake

Two forms of rhodanese were purified from the liver of Clarias gariepinus Burchell, designated catfish rhodanese I (cRHD I) and rhodanese II (cRHD II), by ion-exchange chromatography on a CM-Sepharose CL-6B column and gel filtration through a Sephadex G-75 column. The apparent molecular weight obtained for cRHD I and cRHD II was 34,500 ± 707 and 36,800 ± 283 Da, respectively. The subunit molecular weight determined by sodium dodecyl sulphate–polyacrylamide gel electrophoresis was 33,200 ± 283 and 35,100 ± 141 Da for cRHD I and cRHD II, respectively. Atomic absorption spectrophotometric analysis revealed that cRHD II contained a high level of iron (Fe), which presumably was responsible for the brownish colour of the preparation. In contrast, no Fe was identified in cRHD I, and its preparation was colourless. Further characterization of cRHD II gave true Michaelis–Menten constant (K _m) values of 25.40 ± 1.70 and 18.60 ± 1.68 mM for KCN and Na₂S₂O₃, respectively, an optimum pH of 6.5 and an optimum temperature of 40°C. The Arrhenius plot of the effects of temperature on the reaction rate consisted of two linear segments with a break occurring at 40°C. The apparent activation energy values from these slopes were 7.3 and 72.9 kcal/mol. Inhibition studies on the cRHD II enzyme showed that the activity of the enzyme was not affected by Mn²⁺, Co²⁺, Sn²⁺, Ni²⁺ and NH₄ ⁺, but Zn²⁺ inhibited the enzyme considerably.     

Studies on glutathione transferase from grasshopper (Zonocerus variegatus)

Glutathione transferase (GST) was purified from the hindgut of grasshopper (Zonocerus variegatus) a polyphagous insect. The purified enzyme had a native molecular weight of 40 kDa and a subunit molecular weight of 19 kDa. The purified enzyme could conjugate glutathione (GSH) with 1-chloro-2,4-dinitrobenzene (CDNB), paranitrobenzylchloride, paranitrophenylacetate, 7-chloro-4-nitrobenzo-2-oxa-1,3-diazole (NBDCl), and 1,2-dichloro-4-nitrobenzene (DCNB) with specific activities of 3.3 ± 0.3, 0.49 ± 0.10, 0.10 ± 0.002, 1.2 ± 0.2, and 1.7 ± 0.4 μmol/min/mg protein, respectively. CDNB appears to be the best substrate with a specificity constant, k_cat/K_m, of 1.8 ± 0.1 × 10⁻⁴ M⁻¹ S⁻¹. The kinetic mechanism of Z. variegatus GST (zvGST) in the conjugation of GSH with some electrophilic substrates appears complex. Conjugation of GSH with DCNB was inhibited by high DCNB concentration, while with NBDCl, as the electrophilic substrates, different values of K_m were obtained at high and low concentrations of the substrates. Cibacron blue, hematin, S-hexylglutathione, and oxidized glutathione inhibited the enzyme with I₅₀ values of 0.057 ± 0.004, 0.80 ± 0.2, 33 ± 2 μM, and 5.2 ± 0.3 mM, respectively. The nature of inhibition by each of these inhibitors is either competitive or non-competitive at varying GSH or CDNB as substrates. NADH and NAD⁺ inhibited the enzyme with an I₅₀ value of 0.4 ± 0.01 and 11 ± 1 mM, respectively. NADH at a concentration of 0.54 mM completely abolished the activity. As part of its adaptation, the flexible kinetic pathway of detoxication by zvGST may assist the organism in coping with various xenobiotics encountered in its preferred food plants.     

Effect of Smoking Time and Temperature on the Proximate Composition and Quality of Milkfish Steaks

The effect of three smoking durations (60, 90, and 120 min) and two smoking temperatures (60°C and 70°C) on proximate composition and quality of brackish cultured milkfish steaks of 3 cm size was studied. No significant difference was observed in the sensory properties across all the samples, except in the overall acceptability of sample smoked at 60°C for 60 min. There was significant difference in the values obtained for the textures of all the samples. The L*a*b* values showed a significant difference in the raw and smoked samples. The moisture content showed significant difference in all six samples. Naphthalene, fluorine, and anthracene were detected in the raw sample and in all the smoked samples with significant difference, except for naphthalene. The effect of smoking on the sensory properties and the moisture content of a smoked fish sample are dependent on the smoking time and temperature. Polycyclic aromatic hydrocarbon (PAH) concentration in smoked fish can be a factor, based on both water pollution and the smoking procedure. Benzo-a-pyrene and PAH4, which are acceptable indicators of total PAH presence in smoked foods, were not detected in all the smoked samples, thereby indicating the products to be safe for consumption.     

Status of yam (Dioscorea spp.) in the Democratic Republic of Congo

Yam is an important tuber crop with enormous potential to enhance rural sustenance and livelihood in DRC. However, studies to enhance its genetic improvement are very far from sufficient with only a handful of information available on the crop. Yam has been treated as an orphan crop compared to contemporary crops such as cassava and sweet potato which have adapted to different cropping systems and become widespread in production. The lack of research attention to address the major production challenges has further decreased the value and potential of the crop compared to its contemporaries. These production constraints include lack of adequate quality planting materials, low yield potential, poor resistance/tolerance to yam mosaic and anthracnose diseases and ultimately poor tuber quality attributes focusing on tuber taste, flesh oxidation and dry matter contents of the majority of the farmers preferred varieties. In this review, we evaluated the status of yam in DRC and presented the needful activities to be incorporated for its improvement. Diversity has however been maintained mainly through ennoblement efforts in house gardens and small farmlands using traditional farming methods. Studies from other nations where yam has been successful with prominence in characterization and genetic improvement brought to light the need for DRC to consider yam as a staple carbohydrate food source, even to the extent of modifications in food public policy. Reversal of the yam's current stigma is a challenge to the scientific community and the population in general.     

Investigations of energy metabolism in weanling barrows: the interaction of dietary energy concentration and daily feed (energy) intake

Much of our understanding of energy metabolism in the pig has been derived from studies in which the energy supply was controlled through regulated feed intake. In commercial situations, where ad libitum feeding is practiced, dietary energy concentration, but not daily feed intake, is under producer control. This study evaluated the interactive effects of dietary energy concentration and feeding level (FL) on growth, body composition, and nutrient deposition rates. Individually penned PIC barrows, with an initial BW of 9.5 +/- 1.0 kg, were allotted to 1 of 9 treatments in a 3 x 3 factorial arrangement plus an initial slaughter group (n = 6) that was slaughtered at the beginning of the trial. Three NE concentrations (low, 2.15; medium, 2.26; and high, 2.37 Mcal of NE/kg) and 3 feeding levels (FL: 100, 80, or 70% of ad libitum access to feed) were investigated. Daily feed allowance for the restricted-fed pigs was adjusted twice per week on a BW basis until completion of the experiment at 25 +/- 1 kg of BW. Average daily gain, ADFI, and G:F were unaffected by NE (mean = 572 g, 781 g, and 0.732 g/g, respectively). Average daily gain and ADFI, but not G:F, increased (P < 0.05) with FL. Empty body lipid concentration increased with dietary NE concentration and with FL; a significant (P < 0.01) interaction revealed that empty body lipid concentration increased most rapidly as ADFI increased on the highest energy diet. Empty body lipid concentration was greatest in pigs with ad libitum access to the high-NE diet. Empty body protein concentration decreased with increasing NE (P < 0.05) but was not affected by FL. Empty body protein deposition (PD) increased with increasing FL (P < 0.001), but not with NE. Empty body lipid deposition (LD) and the LD:PD ratio increased (P < 0.01) in pigs with ad libitum access to the high-NE diet. In conclusion, NE did not interact with FL on growth, body protein concentration, or PD, suggesting that the conclusions regarding energy utilization obtained from experiments using restricted feed intake may not easily be applied to pigs fed under ad libitum conditions. The interactive effects of NE and FL on body lipid concentration, LD, and the LD:PD ratio indicate that changes in dietary energy concentration alter the composition of gain without necessarily changing overall BW gain. Consequently, the composition of gain is an important outcome in studies on energy utilization.     

Assessment of the accuracy of estimation of gestational age in cattle from placentome size using inverse regression

AIMS: To assess, in dairy cows between 60–160 days of gestation, whether the agreement between actual gestational age and that predicted by placentome size could be improved by using individual placentome measurements rather than a single average, and to identify how increasing the number of placentomes measured improved the prediction.

METHODS: Data were obtained from 58 cows examined using transrectal ultrasonography every 10 days between 60–130 days, and every 15 days between 130–160 days of gestation. For each cow, four to six placentomes in each of the pregnant and non-pregnant horns were examined from the region of the uterus near to the cervix. A mixed-effects model, which included cow as a random effect, and a simple linear regression which ignored within-cow correlation, were fitted to the data. Inverse regression was used to compare the 95% prediction bands obtained for estimating gestational age using the means of three, five or eight placentome measurements.

RESULTS: The fit of the mixed effects model was better than a simple linear regression (p<0.001) but the fitted lines from the two models were very similar. Using the simple regression model, for a mean placentome length of 15?mm estimated gestational age was 100 days, with 95% CI of 68–131 days for measurement of three placentomes, and 80–120 days for eight placentomes.

CONCLUSIONS: The agreement between placentome size and gestational age was improved by increasing the number of placentomes measured. Direct comparison of these CI with those for other published fetal measures was not possible as similar prediction bands have not previously been calculated for fetal measures; however one study reported a residual SD which had been calculated using size measurements as the predictor variable and age as the response variable. Using these data 95% CI were calculated to be ±9 days for crown-rump length and ±25 days for uterine diameter. These are likely to be an underestimate of the true CI and do not take account of the increase in variance of the difference between predicted and actual gestational age as gestational age increases.

CLINICAL RELEVANCE: Placentome measurement may be useful for estimating fetal age in late gestation. Further research is required to better establish the agreement between gestational age and the fetal measurements which are currently used to estimate fetal age.     

Entertainment-Education Strategy Utilization for Agricultural Information Dissemination: Congruency Among Researchers,Extension Agents,and Farmers in Southwestern Nigeria

Several congruency studies exist on agricultural-related issues, but studies on congruency for entertainment-education (EE) utilization for agricultural information dissemination is limited. Congruency among researchers, extension agents, and farmers on the use of EE for agricultural information dissemination was examined. A multistage sampling procedure was used to select 161 respondents across respondent categories. A structured questionnaire was used to obtain data on respondents' perception and estimate of perception of other stakeholders to establish accuracy. Data were analyzed using Pearson product-moment correlation at p = 0.05. Results indicate congruence between researchers' perception and accuracy in establishing extension agents and farmers' opinion about media environment. A similar result was obtained for congruence between farmers' perception and other stakeholders' position on media environment. Congruence between extension agents, researchers, and farmers was accurate for infrastructural factors. Given the accurate predictions of each other's disposition to key variables, EE should be further exploited for agricultural information dissemination.     

Dietary combination of pawpaw seed and onion peel powder: Impact on growth,haematology, biochemical and antioxidant status of Clarias gariepinus

A feeding trial was conducted to evaluate the effect of pawpaw–onion powder (POP) mixture on the growth, and haemato‐biochemical and antioxidant responses of Clarias gariepinus (4.02 ± 0.01g/fish) for 60 days. Five trial diets were formulated as control (without POP or antimicrobial growth promoter (AGP)), AGP (basal diet + 10ml AGP/kg diet), POP 2.5 (basal diet + 2.5g POP/kg diet), POP 5.0 (basal diet + 5.0g POP/kg diet) and POP 10 (basal diet + 10g POP/kg diet). Two hundred and twenty‐five fish were equally distributed into five groups in triplicate and fed twice daily. The results indicate that AGP or POP supplementation exerted no effects on the growth and blood profile among the various groups, but a significantly higher lymphocyte count was observed in POP 10g/kg. The highest whole‐body protein and lipid contents were noticed in fish fed the control diet (p < .05), whereas POP 5.0g/kg group recorded the highest hepatosomatic value. Furthermore, the cholesterol level was found to be lower in the AGP‐ and POP‐fed fish compared with the higher level recorded in the control. The glucose concentration and superoxide dismutase enzyme activity were found to be higher in POP 2.5‐fed fish, whereas POP 10‐fed fish showed higher catalase activities compared with other groups (p < .05). Based on the result obtained, this study showed that dietary POP had no significant impact on the growth performance but has direct effects on the whole‐body lipid content, lymphocyte count, cholesterol level, alanine aminotransferase, alkaline phosphatase and antioxidant response of Clarias gariepinus.     

Effect of aqueous concentration of humic acid on the sorption of polychlorinated biphenyls onto soil particle grain sizes

Journal of Soils and Sediments - The role of initial aqueous humic acid concentrations (dose), as well as secondary environmental conditions on the sorption of PCB congeners PCB 28, 52, 77, 101,...