Sublethal Effects of Arsenic on Oxygen Consumption, Hematological and Gill Histopathological Indices in Chanos chanos

Background: The current study was performed aiming to evaluate possible changes in the effect on oxygen consumption, hematology and gill histopathological parameters in fish (Chanos chanos) upon exposure to sublethal concentration of the metalloid arsenic. Methods: Bioassay tests were conducted for determining the LC50 values of arsenic for 96 h. Oxygen consumption in control and arsenic-exposed fish was estimated using Winkler’s method. Red blood corpuscular (RBC) count was examined with a Neubauer counting chamber under a phase contrast microscope. Hemoglobin (Hb) was estimated following the acid hematin method. Histopathological studies were carried by processing and staining the gill tissues with hematoxylin and eosin in accordance with standard histological techniques. They were then subjected to examination under a scanning electron microscope. Results: Chanos chanos exposed to 1/10th of LC50 (24.61%) for a period of 30 days exhibited a maximum decline in the rate of respiration, followed by a decline in RBC and Hb above 45.59% and 51.60%, respectively. Significant toxic lesions encompassing fused gill lamellae, detached gill epithelium, hyperplasia and hypertrophy of respiratory epithelium became heavy handed on the 30th day. Conclusion: Information synthesized from our study serves to be useful in monitoring and managing (As) contamination in the aquatic environment.

The Potency of Anadara nodifera Shell as Natural Testosterone Booster for Male Canary (Seriunus canaria)

Anadara clam shell contains high Zinc. This microelement is recognised as acting natural aromatase blocker (NAB) that capable of boosting testosterone level. The aim of this study was to evaluates the testosterone level of Canaries following Anadara nodifera administration. Adult male Canaries (n = 27) were group into 3 with 9 repetition of each i.e T0 [control, was given by Carboxymethyl Cellulose Sodium (Na-CMC)], T1 (Zinc Sulphate 0.013 mg/30 g BW), and T2 (NAB 0.3 mg/30 g BW). The T2 treatment was NAB or shell powder of Anadara nodifera which was diluted 1:1 in Na-CMC. The birds were treated orally for 21 days. The complete blood cell count was performed in a haemo analyser except the erythrocyte was calculated manually in the Improved Neubauer counting chamber. Testosterone level was measured in ELISA. Subsequently gen CYP19 aromatase in syrinx, brain and testis were identified employing IHC and descriptively analysed. The result indicated significant elevated level of testosterone (p<0.05). The expression of CYP19 aromatase receptor was depressed in syrinx, brain and testis. However there was no significant changes on the blood profile (p>0.05). To conclude, Anadara nodifera at 0.3 mg/30 g BW was capable of acting as NAB to promote testosterone level. This was confirmed by the reduction of CYP19 aromatase gene receptors in syrinx, brain and testis. However NAB treatment was not significantly affecting routine blood profile.

A New Approach of Sperm Motility Subpopulation Structure in Donkey and Horse

This study aimed to characterize the sperm kinematic values with high frames per second, to define the subpopulation structure of a horse and a donkey and compare them. A total of 57 fresh semen ejaculates (26 Spanish and 16 Arabian horse breeds and 10 donkeys) were collected and subsequently analyzed for kinematic parameters using the Computer-aided sperm motility analysis ISAS®v1.2 system and using a Spermtrack® 10-μm depth counting chamber. Sequences were recorded at 250 frames per second, and eight kinematic parameters were automatically evaluated. All kinematic parameters showed significant differences between a donkey and a horse and between horse breeds. All ejaculates evaluated showed excellent semen motility characteristics, with significantly higher values for all kinematic parameters for donkeys compared with horses except for beat-cross frequency. Donkey sperm was faster and linear than the horse. Regarding horse breeds differences, the Spanish horse had higher average path velocity, curvilinear velocity, and beat-cross frequency compared with the Arabian horse. Spanish horse sperm was rapid, but Arab horse was more linear. The principal component analysis showed three sperm subpopulations in the ejaculate of donkeys and horses with a significantly different motility characteristic between them. The dominant subpopulation for both donkey and horse was for rapid, straight, and linear with a high beat sperm (38.2 and 41.7%, respectively), whereas the lowest subpopulation was for the slowest and non-linear sperms. This, plus slight differences in the distribution of these subpopulations between Arabian and Spanish horses, were found. In conclusion, higher frames permitted to have a new interpretation of motile subpopulations with species and breed differences. More so, future works on donkey and horse breed spermatozoa should take into account differences between breeds that may interfere and alter the real analysis performed.

78 Evaluation of extended hair sheep ram semen stored as liquid at 5°C

This study was designed to evaluate the quality of extended hair sheep ram semen stored as a liquid at 5°C. St Croix White (STX; n = 6) and Dorper x STX (DRPX, n = 5) rams were collected weekly for 3 wk using estrus ewes fitted with an intravaginal collection vial. Semen was kept at 32°C during transport to the lab and during processing. Semen was evaluated for percent motility (MOT), viability (LIVE) using eosin-nigrosin stain and concentration using a hemocytometer counting chamber. Semen was extended to a final concentration of 250 x 106/mL in a one-step dilution with a skim UHT milk extender with 10% egg yolk by volume and packaged into 0.5 mL straws. Straws were stored in a styrofoam box in a refrigerator at 5°C for 96 h, or in an Equitainer® set up using the manufacturer’s instructions, for 24 h at which time they were transferred to the styrofoam box in the refrigerator for 72 h. Semen was evaluated for MOT and LIVE at -1, 0, 24, 48 72 and 96 h relative to cooling. Semen traits were analyzed using GLM of SAS for repeated measures with ram, time, breed and cooling method in the model. There was no difference (P &gt; 0.10) in MOT or LIVE over time between breeds or cooling method. One STX ram did not produce any samples that survived extension; one STX and four DRPX rams produced at least one sample that did not survive extension and all were removed from analysis. The MOT decreased (P &lt; 0.0001) from 81.7 ± 2.9 % at -1 h to 52.2 ± 2.9% at 96 h. The LIVE decreased (P &lt; 0.0001) from 83.1 ± 3.6% at -1 h to 50.4 ± 3.6% at 96 h. These results show that ram semen stored as a liquid at 5°C can maintain motility and viability for 96 h. Further studies will be conducted to evaluate cooling of extended ram semen and fertility after artificial insemination.

Optimization of CASA-Mot Analysis of Donkey Sperm: Optimum Frame Rate and Values of Kinematic Variables for Different Counting Chamber and Fields

In order to optimize the donkey sperm motility analysis by the CASA (Computer Assisted Sperm Analysis)-Mot system, twelve ejaculates were collected from six jackasses. Capillary loaded chamber (CLC), ISAS®D4C depths 10 and 20 µm, ISAS®D4C Leja 20 and drop displacement chamber (DDC), Spermtrack® (Spk) depths 10 and 20 µm were used. Sperm kinematic variables were evaluated using each chamber and a high-resolution camera capable of capturing a maximum of 500 frames/second (fps). The optimum frame rate (OFR) (defined according to curvilinear velocity—VCL) was dependent on chamber type. The highest OFR obtained was 278.46 fps by Spk20. Values for VCL, straight-line velocity (VSL), straightness (STR), amplitude of lateral head displacement (ALH) and beat cross frequency (BCF) were high in DDC and 10 µm depth. In both DDC 10 and 20 µm, the sperm velocities (VCL, VSL, VAP) and ALH values decreased significantly from the centre to the edges, while Wobble and BCF increased. No defined behavior was observed along the CLC. However, all the kinematic variables had a higher value in a highly concentrated sample, in both chamber types. In conclusion, analyzing a minimum of nine fields at 250 fps from the centre to the edges in Spk10 chamber using a dilution of 30 × 106 sperm/mL offers the best choice for donkey computerised sperm motility analysis.

Coagulation Parameters among Women with Obstetric Complications in Specialist Hospital Sokoto, Nigeria

Background and Aim: Obstetric complications are part of the leading causes of maternal mortality worldwide. This study was carried out to investigate the effect of complications on the Prothrombin time (PT), activated partial thromboplastin time (APTT), and platelet count (PLC). Women with obstetric complications were recruited as subjects). Pregnant women without complications were included as controls. Study Design: This is a case-control study. Place and Duration of Study: The study took place in Obstetrics and Gynaecology Department of Specialist Hospital Sokoto and the duration was six months. Methodology: Pregnant women aged 18–41 years (mean age 29.5 years) were recruited for the study. The determination of PT and APTT was done by manual methods using commercially prepared Agappe reagent kits, whereas PLC was done by manual methods using a haemocytometer (Improved Neubauer counting chamber). Data were analyzed using SPSS version 23. Results: The results of PT and APTT were significantly higher among women with obstetric complications (14.26±0.23 seconds and 31.32±0.70 second) compared to normal pregnant women (13.00 ± 0.13 seconds and 29.66±0.33) (P<0.05). The PLC was significantly lower among women with obstetric complications compared to women without obstetric complications (P< 0.05).Obstetric complications in subject show a significant association when compared with PT and APTT (P< 0.05). The age group of subjects shows a significant association when compared with APTT (P< 0.05). Conclusion: Findings from this study have shown that obstetric complications causes decrease in platelet count and prolongation in PT and APTT. There is need to assess the PT, APTT and Platelet count routinely for pregnant women to improve the ante-natal care in Specialist Hospital Sokoto.

The Method of Total Erythrocyte and Leukocyte Counting in Raptors at Wildlife Rescue Centre (WRC) Jogja

Most of raptor species rehabilitated for conservation purpose at WRC Jogja are eagles. Annually, we perform general raptor examination involving conventional routine hematologic examination. However, low sample quality and over dense blood cell population during total erythrocyte and leukocyte counting were deemed as obstacles. This study aimed to give wider insight in how total erythrocyte and leukocyte counting was performed at WRC Jogja to overcome those obstacles. Firstly, handling and restraint were done physically adapting the Aba method. One-to-two half millilitres of blood was collected from the ulnar vein using 3-ml-syringe with 24-gauge-needle which was then preserved in an EDTA-containing tube and proceeded to subsequent method no more than 3 hours after collection. No hypovolemic shock was observed. During total erythrocyte and leukocyte counting using Neubauer-ruled counting chamber, we modified the dilution factor to be 250-fold and 50-fold respectively. This method yielded a countable cell density and can be applied in other species in similar condition.

Rumus Hitung Eritrosit, Trombosit, dan Leukosit-Improved Neubauer

One (conventional) tool that is still used today to count erythrocytes, platelets and leukocytes is the Improved Neubauer chamber. This method is classified as a classic, but also very cheap, and easy to do in the examination of the count of blood cells. The counting chamber is specifically designed to count the number of erythrocytes, platelets and leukocytes with specific count zones. The area and volume of the zones used to count each cell tend to be different, requiring a separate formula