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Open Access Original Research Article

Assessment of the Conventional versus Two Rapid Microwave Processing Methods Using the Phosphotungstic Acid Haematoxylin Technique

Tobias Peter Pwajok Choji, Anthony Ajuluchukwu Ngokere, Samuel Ifedioranma Ogenyi, Peterside Rinle Kumbish, Lovelyn Peter Unubi, Mada Umoru Alesa, Ponfa Nden Zhakom, Emmanuel Ishaku Vomangai, Gwom Irmiya Davou, Charity Ezikiel Marcus

Annual Research & Review in Biology, Page 1-18
DOI: 10.9734/ARRB/2016/19219

Aims: To process tissues using the rapid microwave techniques and check their reactivity with the phosphotungstic acid haematoxylin. To establish a very fast and cheap method of tissue processing and evaluate its effects on tissue morphology and dye uptake. To eliminate xylene from the processing schedule. To check for additional information that can be obtained from the phosphotungstic acid haematoxylin technique so as to encourage its use in the laboratory.

Study Design: Harvesting and fixation of tissues. Grossing into triplicates, processing using three different techniques. Staining and grading of sections.

Place and Duration of Study: Human samples were obtained from the Anatomy Department, Nnamdi Azikiwe University, Nnewi Campus while animal samples were obtained from the Animal experimental station, National veterinary Research institute, Vom, Nigeria. The work was done between August and December, 2014.

Methodology: Heart, skin, brain, kidney, lungs, and liver tissues were harvested from an apparently healthy rabbit after sacrificing it scientifically. Ethical clearance as well as human cervix, fibroid, prostate and testis were obtained. All samples were fixed in 10% buffered formalin, cut into triplicates and processed using the conventional as well as two rapid microwave techniques respectively. Sections were made from each technique and stained simultaneously using the Phosphotungstic Acid Haematoxylin technique. The Masson Trichrome methos as well as the Haematoxylin and Eosin Techniques were also used to assess morphology They were each evaluated for quality of block, ribboning, nuclear, cytoplasmic and extracellular components, collagen and muscle fibre. Section thickness, uniformity and staining intensities were all checked.

Results: Processing cycle as well as quantity of reagents used was drastically reduced in the microwave techniques. Paraffin blocks obtained, ribbonning as well as flattening on 20% alcohol were comparable in the three techniques. Tissue morphology, integrity of tissue as well as tissue histochemistry to the phosphotungstic acid haematoxylin were comparable among the three techniques. The stroma of the cervix is better retained in the microwave with vacuum technique.

Conclusion: Samples received in histology laboratory can be processed and slides produced within the same day without compromising tissue morphology or histochemistry. Xylene can be eliminated from the paraffin wax processing cycle with quality paraffin block and ribbon production. The cost and duration of tissue processing can be drastically reduced with the production of quality sections. This work can be replicated using other histochemical techniques to give a wider application.

Open Access Original Research Article

Efficacy of Cashew Nutshell Extract against Termite Attack on Triplochiton scleroxylon Wood

A. O. Oladejo, A. Adeleke, F. Musa, W. C. John

Annual Research & Review in Biology, Page 1-7
DOI: 10.9734/ARRB/2016/26066

This study investigated the efficacy of cashew nutshell extract against termite attack on Triplochiton scleroxylon wood in other to add to the quest for readily available and cost-effective pest control materials, specifically on termite attack on wood. The study was carried out at Entomology Laboratory, Federal College of Forestry Jos in Jos North Local Government Area of Plateau state, Nigeria, between April and May 2015. Some prepared wood billets were used as control/untreated (A), while others were treated with cashew nutshell extract in concentrations of 10% (B), 20% (C), 30% (D) and 40% (E) giving all together five (5) treatments which were replicated five (5) times to give a total of twenty five (25) billets. Both treated and untreated wood billets were taken to a selected area that is prone to termite attack by digging holes and placing billets in a Completely Randomized Design (CRD) for eight (8) weeks. After the duration of exposure, wood billets were removed and data was taken for number of termite present at point of removal on each billet, degree of attack, number of furrows and length of furrows and all the data were subjected to the Analysis of Variance (ANOVA) using the SPSS 16.0 package. Latin Square Design (LSD) was further conducted to examine the condition that was responsible for the difference. The result showed that the effects of number of termite present on billets are not significantly different between means. The effect of means percentage on degree of attack showed no significant difference among the treatments 20%, 30% and 40% but showed significant difference with the control and 10%, so also in number of furrows. Observations on the length of furrows showed that control (untreated) was significantly difference from other treatment means. This result led to the conclusion that cashew nutshell extract has pesticidal effects against termite attack on Triplochiton scleroxylon wood with treatment E (40%) having the highest effects.

Open Access Original Research Article

Development, Degradation and Possible Function of Trabecular Trophoblast in the Course of Placentation of Silver fox Vulpes fulvus Desm.

T. G. Zybina, K. M. Pozharisski, G. I. Stein, I. I. Kiknadze, A. I. Zhelezova, E. V. Zybina

Annual Research & Review in Biology, Page 1-9
DOI: 10.9734/ARRB/2016/26057

Aims: Development of trabecular trophoblast cell population in the spongy zone of the endotheliochorial silver fox placenta was investigated with special attention to cytokeratin intracellular arrangement.

Methodology: Cytokeratin immunohistochemistry.

Results: It was shown that keratin filaments contribute to formation of carcass of trabeculae, especially in the zone attached to endometrium In the course of placenta development, in the deep part of spongy zone, there occured progressive detachment of the trophoblast cells out of the trabeculae and acquirement of «migratory» phenotype characterized by specific arrangement of the cytokeratin filaments. Another part of differentiated trabecular trophoblast cells undergo progressive destruction of the cytokeratin filaments that results in complete desintegration of cytoplasm. As a result, a zone filled by products of destruction of the trabecular trophoblast cell is formed between trabeculae and labyrinth.

Conclusion: In the silver fox placentation there are several ways of migration of trophoblast cells that play a role in feto-maternal contacts and in histiotrophic nutrition. First, invasion of trophoblast in the depth of uterine glands with partial replacement of glandular epithelium. Second, detachment of the trophoblast cells into the lumen of trabeculae that is accompanied by acquirement of «migratory phenotype» with specific arrangement of cytokeratin filaments. The nutrients that result from degradation of the trabecular trophoblast cell may be uptaken by the labyrinth, i.e. the main site of supply of nutrient and oxigen to embryo.

Open Access Original Research Article

Integrated Nutrient Management; Option for Improvement on Soil Chemical Properties, Growth and Yield of Cocoyam (Colocasia esculenta)

J. C. Nwite, C. I. Keke, P. O. Okereke

Annual Research & Review in Biology, Page 1-12
DOI: 10.9734/ARRB/2016/25739

The response of cocoyam (Colocasia esuclenta (L.) Schott) and soil chemical properties to different manure sources were studied under field conditions in 2014 and 2015 cropping season at Federal College of Agriculture, Ishiagu, Ebonyi State, a derived savanna zone of Southeastern Nigeria. Randomized complete block design (RCBD) was used to study these soil amendments. Ten(10) treatments; 10t/ha poultry dropping (PD), 200 kg/ha NPK 15:15:15 fertilizer (NPK), 5 t/ha rice husk ash (RHA), 10 t/ha rice husk dust (RHD), 5t/ha PD + 2.5 t/ha RHA, 5 t/ha PD + 5 t/ha RHD, 100 kg/ha NPK + 2.5 t/ha RHA, 100 kg/ha NPK + 5t/ha RHD, 66.67 kg/ha NPK + 3.33 t/ha RHD + 1.67 t/ha RHA and control, were used in the study which were replicated three times. Soil chemical properties evaluated were pH, organic carbon, exchangeable bases (K+, Ca2+, Mg 2+ and Na+) and exchangeable acidity. Others include CEC, total nitrogen, available phosphorus and base saturation while cocoyam yield was measured at harvest. The results showed that soil pH, soil organic carbon (SOC), cation exchange capacity (CEC), exchangeable acidity (EA), BS and available phosphorous were significantly affected by the application of the amendments, while total nitrogen shows non-significant effect. Cocoyam cormels and corms yield were significantly (p < 0.05) increased in all the soil amended plots than the control. Generally, the results implied that the integration of organic and inorganic amendments stand better chance of improving soil fertility indices and the crop yield than their sole forms in the study.

Open Access Original Research Article

Evaluation of Dietary Supplementation of Ammonium Sulphate on in vitro Gas Production and Rumen Fermentation Characteristics of WAD Ewes

T. O. Ososanya, S. A. Shehu, U. A. Inyang

Annual Research & Review in Biology, Page 1-7
DOI: 10.9734/ARRB/2016/25775

Ammonium sulphate, a chemical compound was utilized to evaluate its potential through in vitro gas production technique and rumen fermentation characteristics as a rumen stimulator and mitigates methane gas. Four experimental diets were formulated with ammonium sulphate (AS) included at varying levels: T1 (control/0% AS), T2 (control diet + 0.25% AS), T3 (control diet + 0.50% AS) and T4 (control diet + 0.75% AS). Rumen fluid was collected from the ewes, sixteen in number and weighing 24 kg averagely, using suction tube method to evaluate in vitro gas production, determine microbial population and rumen fermentation characteristics. Gas production was determined over a 96 hour period. Other data collected included organic matter digestibility (OMD%), short Chain Fatty Acids (SCFA µml), Metabolisable Energy (ME MJ/KJ DM), Degradability (D%), Volatile Fatty Acids (VFA), rumen pH, temperature and ammonia nitrogen (AN). Results showed no significant differences (p>0.05) for in vitro gas production profiles (i.e. total gas, OMD, ME, SCFA). However, as inclusion increased it stimulated the parameters measured to be numerically higher than control. Methane gas was highest for control diet (4 ml/200 mg DM) while T2 recorded the least (2 ml/200 mg DM). Degradability increased as the inclusion level of AS increased. Control diet was the lowest (32%) while T4 had the highest (45%). The rumen fermentation characteristics showed significant (p<0.05) differences for all the parameters measured except rumen temperature. Animals on T4 diet recorded the highest (p<0.05) pH value (6.85) while those on T2 diet had the least (6.53). Animals on Control diet recorded the highest values (p<0.05) for AN and acetic acid (100.40 mg/l and 8.98 mol/100L) while the least values were observed in animals on T2 diet (AN) and those on T4 (acetic acid). Increase inclusion of AS caused a decrease in acetic acid. Rumen bacteria was highest (p<0.05) in animals on T4 (1.40 x 106cfu/mm3) and lowest in those on T3 (0.65 x 106cfu/mm3). For fungi and protozoa, animals on T4 and T2 diets were higher (p<0.05) respectively than those on Control diet (1.15 and 1.09 x 106cfu/mm3  compared to 1.10 and 0.85 x 106cfu/mm3 respectively). Supplementation of AS has been validated by this study, with a 0.75% inclusion level resulting in lowest methane gas, and higher ME, SCFA, pH, bacteria and fungi and may be recommended for ruminant feeding.

Open Access Original Research Article

Gene Sequencing and Haemolysis of Bacteria in Clarias gariepinus From Ajilete Location on Yewa River

O. O. Ololade, O. O. Oyelakin, A. R. Oloyede, A. A. Idowu, A. A. Akinyemi, Y. A. Babarinde

Annual Research & Review in Biology, Page 1-8
DOI: 10.9734/ARRB/2016/26923

Gene sequencing and haemolysis of bacteria in Clarias gariepinus post juveniles sampled from Ajilete location on Yewa River, Ogun state, Nigeria was carried out. Bacteria were isolated from the gut, gills and skin of the fish. Thirty bacteria isolates were selected and the DNAs were extracted using hexadecyltrimethylammonium bromide (CTAB) method and Polymerase Chain Reaction (PCR) amplification of the isolates was carried out using universal primer for bacteria. The purification of PCR product was done using ethanol precipitation and thereafter the DNAs were sequenced using automated DNA sequencer. The sequence data were analyzed on the GenBank database (NCBI) using Basic Local Alignment Search Tool (BLAST) to find the nearest related sequence, and to construct the phylogenetic relationship. The haemolysis of the selected isolates were also carried out. Data collected were subjected to descriptive statistics. Nine genera of bacteria isolated were Escherichia, Staphylococcus, Enterobacter, Proteus, Pseudomonas, Klebsiella, Citrobacter, Streptococcus and Bacillus. The 16S rRNA gene sequencing method identified the isolates as strains of Brevibacillus agri, Bacillus cereus, Ignatzschineria indica, Comamonas kerstersii, Proteus vulgaris, Lysinibacillus sphaericus, Proteus hauseri, Clostridium bifermentans, Proteus mirabilis, Pseudomonas putida, Escherichia coli, Comamonas jiangduensis, Enterobacter cloacae, Serratia marcescens, Alcaligenes faecalis, Lysinibacillus fusiformis, Myroides odoratimimus. Haemolysis showed that M. odoratimimus, B. cereus, C. kerstersii, P. vulgaris, C. bifermentans, E. coli and E. cloacae all had β-haemolysis. A. faecalis displayed α-haemolysis while, B. agri, I. indica, L. sphaericus, P. hauseri, L. fusiformis, S. marcescens were non-haemolytic. This study confirms the reliability of the 16S rRNA gene sequencing method. This study also concludes that there are different pathogenic bacteria species that are associated with C. gariepinus.

Open Access Original Research Article

Cellular and Humoral Immune Response in Vertically Transmitted Salmonella isolates in Broiler Chickens: A Case Study of Ogun, Oyo and Lagos States, Nigeria

R. A. Olorunsola, P. A. Akinduti, F. O. Oke, A. O. Akapo, A. O. Oso, D. Eruvbetine

Annual Research & Review in Biology, Page 1-8
DOI: 10.9734/ARRB/2016/26965

This study investigated the humoral and cellular immune response in vertically transmitted salmonellaisolates in broiler chickens in some selected states of South-Western Nigeria and its control through the use of feed additives. Anak 2000 day-old broiler chickens totaling 360 (120 birds from each state) were collected from hatcheries that were positive to Salmonella organisms and used for performance testing which lasted for 8 weeks. The birds were laid out in a 3x5 factorial arrangement comprising of 5 dietary treatments including a control and 4 different feed additives (mannose oligosaccharide (MOS), arabinoxylose oligosaccharide (AXOS) and Pediococus acidilactici (SIML) and antibiotic; Oxytetracycline). Blood samples were collected from the birds and their total and differential white blood cells were determined while humoral antibody titre to Salmonella was performed.

Total white blood cells (TWBC) was significantly (P<0.05) increased in the control diet across the locations compared to lower values in other dietary treatments applied. Neutrophil was significantly (P<0.05) increased across the location and decrease with various dietary treatments while Lymphocyte and Basophil values varied significantly (P<0.05) across the locations and the treatments. Monocyte and Eosinophil were not affected by both location of the hatchery and additives. Salmonella antibody titre ≤ 1:20 for ‘O’ and ‘H’ antigen was observed in all birds from various location after treatment with antibiotic (oxytetracyline) and significant reduction of salmonella antibody among birds fed with MOS, AXOS and SIML.

Inclusion of probiotic and prebiotic additives are effective and safe methods for prevention of Salmonellainfection in broiler chickens and enhance poultry productivity.