Commercial Diluent Research Articles

Effects of in ovo injection of carbohydrates on somatic characteristics and liver nutrient profiles of broiler embryos and hatchlings ,

Effects of the in ovo injection of commercial diluent supplemented with dextrin or with dextrin in combination with various other carbohydrates on the somatic characteristics and liver nutrient profiles of Ross × Ross 708 broiler embryos and chicks were investigated. Results include information concerning the gluconeogenic energy status of the liver before and after hatch. Eggs containing live embryos were injected in the amnion on d 18 of incubation using an automated multiple-egg injector for the delivery of the following carbohydrates dissolved in 0.4 mL of commercial diluent: 1) 6.25% glucose and 18.75% dextrin; 2) 6.25% sucrose and 18.75% dextrin; 3) 6.25% maltose and 18.75% dextrin; and 4) 25% dextrin. Also, a noninjected control and a 0.4-mL diluent-injected control were included. Body weight relative to set egg weight on d 19 of incubation (E19) was increased by the injection of all carbohydrate solutions, and on the day of hatch was increased by the injection of diluent, sucrose and dextrin, and maltose and dextrin solutions. Hatchability of the fertilized eggs, residual yolk sac weight, and liver weight were not affected by any injection treatment; however, as compared with the 0.4 mL diluent-injected group, all of the supplementary carbohydrates, except for the glucose and dextrin combination group, increased liver glycogen and glucose concentrations on E19. Furthermore, all carbohydrates, except for the 25% dextrin treatment, decreased liver fat concentration on E19. From E19 to the day of hatch, liver glycogen concentrations dropped dramatically from an average of 3.2 to 0.6%. Despite treatment differences observed on E19 for liver glycogen, glucose, and fat concentrations, these differences were lost by the day of hatch. Nevertheless, liver glycogen and glucose concentrations were positively correlated on the day of hatch. In conclusion, the in ovo injection of various supplemental carbohydrates dissolved in 0.4 mL of commercial diluent altered the liver nutrient profile of Ross × Ross 708 broiler embryos before hatch. However, the subsequent pattern of energy utilization during the hatching process modified these effects.

Effects of commercial in ovo injection of carbohydrates on broiler embryogenesis ,

The effects of in ovo injection of different carbohydrate solutions on hatchability of fertilized eggs (HF), rate of hatch, BW, body moisture, yolk sac weight, and yolk sac moisture of Ross × 708 broiler chicks, hatched from eggs laid by a 34-wk-old breeder flock, were investigated. Eggs containing live embryos were injected, using an automated multiple-egg injector, in the amnion on d 18.5 of incubation with 0.1, 0.4, 0.7, or 1.0 mL of commercial diluent or a carbohydrate dissolved in diluent. The commercial diluent containing 0.25 g/mL of one of the following carbohydrates was injected into eggs: glucose, fructose, sucrose, maltose, or dextrin. The results showed that no carbohydrate type or solution volume affected rate of hatch. Absolute and proportional BW on day of hatch were positively related to injection volume (P < 0.001). However, HF was negatively related to injection volume (P < 0.001). To realize an HF of 90%, the injection volume could not exceed 0.4 mL for fructose or sucrose and could not exceed 0.7 mL for glucose, maltose, or dextrin. Yolk-free BW was negatively related to injection volume of fructose and sucrose (P < 0.004), but was not related to injection volume of diluent, glucose, maltose, and dextrin. Conversely, absolute and proportional yolk sac weights were positively related to injection volume of fructose, sucrose, and dextrin (P < 0.01), but were also not significantly related to injection volume of diluent, glucose, and maltose. Yolk sac moisture was positively related to injection volume for all injectables, including the diluent (P < 0.03). However, body moisture and yolk-free body moisture were not related to injection type or volume. In conclusion, the use of carbohydrates added to a commercial diluent for the in ovo injection of broiler hatching eggs requires the use of appropriate volumes to promote growth and nutrient utilization without adversely affecting HF.

Effects of the utilization of homeopathic elements in commercial diluent on swine sperm viability

It has been speculated that the homeopathic treatment of sperm cells in order to improve semen quality could be promising. However, few data is available and its use in spermatozoa requires investigation. It is well established that mitochondrial membrane potential is an important viability parameter of spermatozoa and it is intimately related to reproductive efficiency. In this manner, new technologies in order to improve the activity of sperm cells and, finally, the fecundity of swine herds are of extremely importance. Due to the lack of knowledge of homeopathic treatment effect on spermatozoa, the aim of the present study was to verify the effect of three different homeopathic treatments on viability of boar sperm cells. Three homeopathic treatments composed by Pulsatila CH6, Pulsatila and Avena CH6, Avena CH6 and one control treatment (sucrose) were added to diluted boar semen, which were cooled for 24 or 48 h. Interestingly, no positive effect of homeopathic treatments was observed over semen viability. However, it was demonstrated that the 24 h of cooling storage provided more viable sperm cells when compared to the 48-h period. This effect of storage period on sperm viability was assessed by intact plasmatic membrane, intact acrosome and mitochondrial membrane potential evaluation.

Elimination of Apoptotic Spermatozoa from Rabbit Insemination Dose Using Annexin V Associated with the MACS Technique. A Preliminary Study

The aim of this study was to verify whether the separation and elimination of the apoptotic fraction in rabbit semen using a MACS technique may improve sperm fertility potential and consequently rabbit kindling rate. Semen samples from 25 New Zealand White (NZW) rabbit males were collected using an artificial vagina and evaluated using the CASA system for concentration and motility. For artificial insemination the best 11 bucks were chosen based on motility parameters. Their ejaculates were mixed to make a heterospermic pool and routinely diluted in a commercial insemination diluent (MiniTüb, Tiefenbach, Germany) at a ratio of 1:6. Diluted heterospermic spermatozoa were filtered through a Sartorius filter to wash out seminal plasma, re-diluted in binding buffer (Annexin V Microbead Kit, Miltenyi Biotec, Germany) at a ratio of 1:3.66 and divided into two groups: an experimental group intended for MACS separation and control group without MACS separation. Then hormonally treated females of NZW rabbits were inseminated with fresh doses of filtered heterospermic semen (n = 27; 0.5 ml I.D. per female) and MACS separated semen (n=28; 0.5 ml I.D. per female). Separation and subsequent elimination of apoptotic spermatozoa (positive selection) from the insemination dose (after negative MACS selection) was verified under in vivo conditions on the basis of increased kindling rate in the experimental group in comparison with kindling rate in the control group (81.3% vs. 73.8%). In conclusion, elimination of apoptotic spermatozoa by the use of the MACS technique results in a slight improvement in kindling rate of rabbit does.

Evaluation of Antibody Diluents for FANCD2 Immunohistochemistry with Breast Cancer Tissue

Two important criteria used to select an antibody diluent for robust immunohistochemistry are: (1) optimal signal/noise ratio, and (2) long-term stability when diluted. For immunohistochemistry with Fanconi anemia, complementation group D2 (FANCD2) in formalin-fixed-paraffin-embedded breast cancer tissue, eight experimental and eight commercial diluents were evaluated. Staining intensity was quantitated with Aperio digital image analysis software. An H-Score was calculated for FANCD2 and isotype control staining for each diluent. Signal/noise ratio was defined as H-score of the antibody divided by H-score of the isotype control. The ratios ranged from 0.46 to 135.9 for different diluents. Casein in Tris-based buffer produced the greatest signal/noise ratio. The two best commercial and four best experimental diluents were selected for evaluation of the diluted FANCD2 antibody stability at 4°C, room temperature, and 37°C, respectively. Three diluents were eliminated from further investigation after 2 weeks because of suboptimal staining. Evaluation of room temperature and 37°C storage ended at 1 month, and evaluation of 4°C storage ended at 6 months. FANCD2 antibody was stable for at least 1 month at room temperature and 37°C, and for at least 6 months at 4°C in casein-based diluents, as well as the diluent containing bovine serum albumin and normal horse serum.

Effects of in ovo injection of L-carnitine on hatchability and subsequent broiler performance and slaughter yield1, 2

Effects of in ovo injection of L-carnitine on the hatchability, grow-out performance, and slaughter yield of Ross x Ross 308 broilers from a young breeder flock were determined through 48 d of age. Fertilized eggs were injected in the amnion with L-carnitine (0.5, 2.0, or 8.0 mg dissolved in 100 microL of a commercial diluent) on d 18 of incubation using an automated egg injector. Three control groups (noninjected and injected with or without diluent) were also included. Hatchability and hatch rate of fertilized eggs were assessed. Furthermore, subsequent mortality, BW gain, feed intake per bird, and feed conversion were determined through 46 d posthatch. On d 47, live body, carcass, and abdominal fat pad weights, along with the weights of all major commercial cuts including the thigh, drumstick, wings, and breast muscles, were determined. Individual doses of supplemental L-carnitine had no significant effect on the hatchability or rate of hatch of fertilized eggs; however, significant trends were noted for increased hatchability and length of egg incubation in conjunction with increases in L-carnitine dose. Nevertheless, there were no significant treatment effects on any of the grow-out performance or slaughter yield parameters investigated. In conclusion, although increasing the levels of L-carnitine added to commercial vaccine diluent between 0.5 and 8.0 mg/100 microL for commercial in ovo injection did not significantly influence subsequent broiler grow-out performance or slaughter yield, L-carnitine dosages above those used in this study have the potential for significantly increasing incubation length and hatchability of broiler hatching eggs.

EFFECT OF DILUENT AND STORAGE TIME OF RABBIT SEMEN ON THE FERTILITY OF DOES REARED UNDER TWO DIFFERENT LIGHTING SCHEDULES

Using a 3-factorial design, the effect of two commercial diluents (IMV vs Minitüb), two storage times (fresh vs 6h storage at ambient temperature) and a lighting schedule was studIed on artificial inseminated does. During a 10 month experimental period 140 (nulliparous and multiparous) does were randomly divided in two production groups and submitted to a 16h r :8hD continuously lighting schedule or to a 10hL:14hD schedule. In order to synchronize the oestrus, the lighting period was suddenly increased till 16hL from 5 days before the insemination (alternative lighting). Both production groups were inseminated every a weeks with an interval of 3 weeks between them. In total 988 inseminations were performed with an average conception rate (CR) of 56.5 %. The CR with both diluents was comparable, 55.6 % and 57.5 % for IMV and Minitüb, respectively. There was no decrease observed n CR when semen was stored during 6 hours into the insemination straws. CR was respectively 56.6 % (fresh semen) and 56.4 % (stored semen). The difference in CR between lighting schedules was small (2 %) and also not significant. Differences in litter size were not significant and could not be relatad to the treatments used. The overall low CR (56.5 %) is partly explained by the physiological status (vulva colour) of the does, indicating that 67 % of the does were not receptiva (white or rose vulva) at the 1 st insemination. However, on does showing a red or purple vulva a CR of 70.3 % was obtained. â–¡tter size of does inseminated with white vulva was significantly (P&lt;0.001) lower, and amounted only 3.9 (nulliparous does) and 5.8 (multiparous does) live youngs instead of 6.9 (nulliparous, rose vulva) to 8.9 (multiparous, purple vulva) when inseminated with coloured vulva.

Viabilidade in vitro de células espermáticas ovinas submetidas a diferentes diluentes e a refrigeração

Avaliou-se a viabilidade in vitro de células espermáticas após a adição de três diluentes comerciais, que foram comparados com o diluente tradicional Tris-gema, utilizados no processo de refrigeração do sêmen ovino, em até 48h de armazenamento. Foram utilizados nove ejaculados diários, obtidos de três reprodutores Dorper, com vagina artificial, em três repetições com intervalo de três dias. O sêmen foi mantido a 30°C, e foram avaliadas suas características macro e microscópicas. Após formação do pool, repetiram-se as avaliações acrescidas da concentração espermática e da integridade do DNA e do acrossoma. Dividiu-se o pool em cinco tratamento, cada um constituído de uma parte de sêmen para três partes dos respectivos diluentes: Equimix (DI), Laiciphos Green Ovine (DII), FR 4 (DIII), Equimix-Gema- Equimix com gema de ovo (DIV) e Tris-Gema (DV). O material obtido em cada tratamento foi subdividido em quadruplicata, refrigerado e mantido a 4°C até as avaliações da motilidade individual progressiva (MIP), do vigor e da integridade do DNA e do acrossoma, correspondendo a zero, 12, 24, 36 e 48h. Nas avaliações do sêmen, com o DI ocorreu a maior queda de MIP às 12h em relação aos demais grupos (P&lt;0,05). Às 24h, os tratamentos DII, DIV e DV apresentaram a melhor MIP (P&lt;0,05), que não divergiram (P&gt;0,05) entre si; às 48h, o DII e o DV foram melhores (P&lt;0,05) que os demais. Com relação ao vigor, os tratamentos DII e DV apresentaram valores mais elevados (P&lt;0,05) em relação ao DI e DIII, a partir das 12h, e ao DIV, a partir das 24h (P&lt;0,05). Concluiu-se que o diluente Laiciphos Green Ovine, da mesma forma que o Tris-gema, pode ser utilizado na conservação do sêmen a 4°C por 48h, enquanto o Equimix, acrescido de 20% de gema de ovo, pode ser seja utilizado no armazenamento do sêmen (4°C) por até 24h.

Developing of Tris Soy Milk Diluent for Frisian Holstein Bull Frozen Semen

® (Minitub, Germany). Since the commercial diluents were imported, they were often not readily available. This research aimed to develop Tris-soy modified diluent, and investigated its effects on the quality and fertility of Frisian Holstein (FH) bulls frozen semen. This research consists of two experiments. At first, we compared the FH Bull frozen semen quality in modified tris soy milk (TSM), Andromed ® and Tris-eggyolk (TEY) diluents, the second was the fertility trials of the frozen semen. Results of the experiment demostrated that post thawed sperm motility in the semen preserved with TEY (49.10%) or Andromed ® (50.21%) was significantly higher (P = 0.037) than that preserved with TSM (41.53%). In contrast, the conception rates in cows inseminated with semen preserved with TSM, TEY, and Andromed ® were 53.84% (7/13), 38.88% (5/13), and 38.46% (7/19), respectively. We are optimistic that TSM diluents will have similar qualities as TEY and Andromed ® on preserving frozen semen by doing future intensive studies.

Combustion synthesis of single-phase β-sialons ( z = 2–4)

Single-phase β-sialon powders ( z = 2–4) have been prepared with homogeneous compositions by the combustion synthesis. The raw materials (Si, Al and SiO 2) were combusted under N 2 pressure of 1 MPa. Without using a diluent, the reaction temperatures were very high (>2000 °C) and the combustion products contained Si and Al residues. With addition of commercial β-sialon ( z = 1) as a diluent (up to 50 wt%), both the reaction temperatures and amount of residual Si and Al significantly decreased. The combustion reactions completed at 50 wt% dilution, and pure β-sialon phases were synthesized. When the combustion product itself is used instead of the commercial diluent, the phase content of desired z value increased with the repetition times until a single-phase powder is produced. The sinterability of the synthesized powders was then tested using 5 wt% Y 2O 3 as a sintering aid by the spark plasma sintering (SPS).

Effects of in ovo injection ofl-carnitine on subsequent broiler chick tissue nutrient profiles

Effects of in ovo injection of L-carnitine on BW and the moisture and nutrient biochemical concentrations of various organs and muscles of Ross x Ross 308 broiler chicks, hatched from eggs laid by a 28-wk-old breeder flock, were determined through 48 d posthatch. Eggs containing live embryos were injected in the amnion with L-carnitine (0.5, 2.0, or 8.0 mg dissolved in 100 microL of a commercial diluent) on d 18 of incubation using an automated egg injector. Three control groups (noninjected and injected with or without diluent) were also included. On d 0, 3, 10, 28, and 48 posthatch, bird BW and the proportional weights and moisture concentrations of various organs and muscles were determined. Glycogen, glucose, protein, and fat concentrations were also determined in certain tissue samples. Bird BW; proportional liver weight; breast, thigh, and gastrocnemius muscle moisture; liver glycogen, glucose, and protein concentrations; and breast and thigh muscle fat and protein concentrations changed with posthatch bird age. Liver glucose on d 0 and pipping muscle moisture on d 3 posthatch were significantly affected by treatment. In comparison to eggs injected with commercial diluent with no added L-carnitine, liver glucose was reduced by the injection of diluent containing either 0.5 or 8.0 mg of L-carnitine, and pipping muscle moisture was increased by the injection of commercial diluent containing either 0.5 or 2.0 mg of L-carnitine. The modified concentrations of the 2 parameters in response to these treatments were not different from those in noninjected control eggs. In conclusion, L-carnitine added to commercial vaccine diluent at levels between 0.5 and 8.0 mg/100 microL for the commercial injection of broiler hatching eggs may decrease liver glucose and increase pipping muscle moisture concentrations of chicks on d 0 and 3 posthatch, respectively, so that their levels are commensurate with noninjected controls.

Tipos e volumes de diluentes para inseminação artificial intrauterina em suínos

A pesquisa foi conduzida com o objetivo de avaliar diferentes tipos e volumes de diluentes por meio do desempenho reprodutivo de 72 fêmeas suínas inseminadas artificialmente. Utilizou-se o delineamento em blocos ao acaso, em esquema fatorial 3 × 2, com três diluentes e dois volumes inseminantes. O sêmen utilizado foi obtido de três machos de fertilidade comprovada. Os ejaculados foram diluídos em partes iguais com os diluentes comerciais A, B e C, constituindo doses com 1,5 bilhão de espermatozoides em volumes de 15 ou 30 mL. As variáveis analisadas foram taxa de parto, índice de retorno ao estro, número total de leitões nascidos, número de leitões nascidos vivos, mumificados e natimortos e peso da leitegada ao nascimento. Não foram observadas diferenças entre as taxas de parto e as taxas de retorno ao estro. As maiores médias de número total de nascidos, nascidos vivos e peso de leitegada foram de fêmeas inseminadas com doses contendo os diluentes B e C. Não houve diferenças entre os volumes inseminantes dentro dos diluentes B e C para nenhuma das variáveis. Para o diluente A, o volume que proporcionou maior tamanho de leitegada e número de leitões nascidos vivos foi o de 15 mL. O número de leitões natimortos e mumificados não diferiu entre os tipos e volumes de diluentes. A inseminação intrauterina de fêmeas suínas de ordem de parto 2 a 5 com doses inseminantes de 15 mL diluídas em diluentes de curta duração (diluentes B ou C) permite índices de desempenho reprodutivo satisfatórios.

Design of Steam-Hexane Injection Wells For Gravity Drainage Systems

Abstract The steam-assisted gravity drainage (SAGD) process has been demonstrated to be commercially viable for the recovery of bitumen from the Athabasca Oil Sands region. One of the potential long-term concerns with the process is the relatively high use of energy. The majority of the projects underway have steam-oil ratios (SOR's) in the range of 2.5 to 3.5. This translates into combustion of approximately 20 – 28% of the energy contained in the oil produced. To reduce the current level of energy consumption, many studies have investigated the addition of solvents to the steam. A small number of field trials have been conducted, with apparently encouraging results. The use of hexane, or a similar commercial grade naphtha mixture, as a solvent appears promising, as it would closely match the vapour pressure curve of steam and would tend to flow and condense with the steam. This study presents the results of numerical investigations regarding the implications of the addition of hexane to steam with respect to the design of SAGD solvent injection wells. This study found that the SOR at full depletion of a steam-hexane SAGD, in the absence of a wind-down strategy, would be improved over that for conventional SAGD only by the degree that heat losses to the over- and underburden are reduced due to more rapid bitumen production rates. At short and intermediate times, a more significant SOR improvement will result primarily from the more efficient drainage of bitumen from the chamber walls and floor, reducing the thickness of the heated formation that is undrained. It was also concluded that injection wells for steam-hexane SAGD will require the same diameter wellbores as for conventional SAGD injection wells. Existing SAGD injection wells can be directly converted to steam-hexane use as overall project economics dictate. Introduction The SAGD process has now been demonstrated to be a robust technique to produce bitumen from high quality reservoirs that are too deep to be mined economically from the surface. There are a significant number of commercial projects at the time of writing, with many more in the planning stages. However, it is well-known that the SAGD process is very energy intensive. A typical high quality SAGD project exhibiting a wellhead SOR of 2.5 will require the consumption of ~20% of the bitumen produced to generate steam (based on 80% conversion efficiency in the boiler). There have been many authors(1–10) that have suggested that the addition of solvents to steam might be a method to improve the energy efficiency of the process and also to potentially increase the production rate. If successful, such steam-solvent processes would be economically beneficial. A laboratory investigation by Nasr et al.(1) showed that drainage rates with the injection of a steam-hexane mixture with 1% mass fraction hexane produced average drainage rates that were approximately double those obtained for a steam only system. This same study showed that steam-hexane produced the highest drainage rates of the solvents tested (C1 through C8, plus a commercial diluent).

Cryopreservation of dog semen: The effects of Equex STM paste on plasma membrane fluidity and the control of intracellular free calcium

Understanding cryoinjury of dog spermatozoa is crucial to preserving fertilizing ability. This study examined flow cytometric indicators of sperm function to explore the reported benefits of Equex STM paste. The motility of cryopreserved spermatozoa immediately and 1 h after thawing was higher in the extender containing 0.5% Equex; no significant differences between the two extenders were observed regarding viability, acrosomal integrity and intracellular Ca 2+ concentration. The proportion of spermatozoa having high membrane fluidity increased significantly post-thawing. The interaction between time after thawing and treatment was significant for plasma membrane fluidity. Dilution in a commercial diluent for transport before processing caused a significant increase in intracellular Ca 2+, which may affect functional survival. No significant difference with or without Equex was detected in plasma membrane fluidity. However, a significant interaction between Equex and dogs was detected. A significant decrease in intracellular Ca 2+ was detected in the live cell population both after dilution in Andersen's buffer and again after cooling and equilibration. One hour post-thaw, the proportion of live spermatozoa with high calcium concentration increased to a similar proportion as that seen in diluted semen; the interaction between diluent and dog was significant. The results suggest that Equex in the diluent benefited motility after cryopreservation. Live spermatozoa with high intracellular Ca 2+ after cryopreservation seem to have a favoured survival in the first hour after thawing. Nevertheless, survival after cryopreservation was severely compromised, explaining the relatively poor fertility of cryopreserved dog semen.

Sperm cryoconservation in Anarhichas sp., endangered cold-water aquaculture species with internal fertilization

This study contributes to the identification of an adequate cryopreservation medium for wolffishes sperm (Anarhichas minor and A. lupus). Our results also confirm the presence of antifreeze proteins in the seminal fluid of the Atlantic wolffish (Anarhichas lupus) that could facilitate the process of cryoconservation in association with increased post-thaw motility and fertilization rates. Cryo-Fish, a commercial diluent was the most efficient in our trials, with 80% motility post-thaw.

Application of Modified Natural Oils as Reactive Diluents for Epoxy Resins

AbstractBisphenol A based low‐molecular‐weight epoxy resin was modified with epoxidized soybean oil, which exhibit viscosity reducing ability comparable to commercial grade active diluents. The studied compositions showed a non‐Newtonian rheological behavior, typical for Bingham liquids. The values of the flow index (n) and the consistency index (k) for the compositions tested in the temperature range 25–65 °C were calculated from the Ostwald‐de Waele rheological model and were used to calculate the flow‐activation energy (Ea) using the Arhenius equation. Studies of co‐crosslinking of mixed oil‐resin compositions using isophorone diamine showed essential decrease of the reaction heat and peak maximum temperature. Mechanical properties, thermal stability, water absorption and chemical resistance of the epoxy resin modified with natural oil, were also investigated. Compositions of epoxy resin Ruetapox 0162, modified with the oil diluent, preserved very good mechanical properties of the epoxy resins and demonstrated relatively low water absorption as well as high chemical resistance. The compositions displayed even higher impact strength than pure epoxy resin due to plasticizing effect of the built‐in oil. Compositions with the high contents (up to 60 weight %) of the oil were flexible materials with fast elastic recovery.

Direct electrochemical detection of sodium azide in physiological saline buffers using highly boron-doped diamond electrodes

Direct detection of sodium azide was examined by using highly boron-doped diamond (BDD) electrodes in some physiological saline buffer solutions. Three important saline buffers, ADA buffer ( N-(2-acetamido) iminodiacetic acid), sodium EDTA and imidazole, were investigated. Hydrogen-terminated BDD was fixed for the investigation since it showed much better sensitivity for azide oxidation in comparison with the oxygen-terminated one. Wide range of buffer concentration was examined. A well-separated peak of sodium azide was observed in its mixture with the buffers, resulting in selective sodium azide detection. The current responses were exactly total addition of sodium azide and buffer current responses. The present method was evidenced to be available for direct detection of sodium azide in two samples of commercial diluents for cell counting. A standard addition method was applied. Sodium azide concentrations of 502 and 1355 μg mL −1 were detected in both samples with the R.S.D.s of 0.07 and 2.37%, respectively. Validation was achieved by the comparison with the sample concentrations given by the manufacturer. Simple, selective, good sensitivity and excellent reproducible detection make the diamond electrodes very attractive for numerous electroanalytical applications.

Detection of red cell antibodies: comparison of two low ionic strength diluents

Various low ionic strength diluents are used routinely for red cell alloantibody detection in the antiglobulin test to increase the rate of antibody association to antigen, thereby allowing a reduction in the incubation time while achieving optimal agglutination. Two commercial low ionic strength diluents (DiaMed ID-CellStab and Inverclyde LISS) were assessed using the DiaMed-ID LISS Coombs’ microtube column system, to assess whether or not the choice of diluent influences red cell antibody detection. Effects of two low ionic strength diluents after 15-min incubation were assessed in 150 samples containing a wide range of typical red cell alloantibodies. Inverclyde LISS gave significantly higher reaction strengths in 25% of samples when compared with the same red cells suspended in ID-CellStab. Variation in reaction strengths ranged from 1+ to 2+, using Inverclyde LISS versus CellStab. Of 131 red cell alloantibodies directed against Rh, Kell, Kidd and Duffy antigens, Inverclyde LISS detected 90% after 15-min incubation, whereas 83% were detected with CellStab. This study suggests that Inverclyde LISS provides better red cell alloantibody detection than does ID-CellStab, and this may be due to the higher ionic strength of ID-CellStab.

Conservation of fresh ram spermatozoa at 5°C in the presence of seminal plasma

Seminal plasma aids sperm transport and contains factors beneficial for sperm function. In artificial insemination, however, diluting the semen reduces the concentration of seminal plasma. To test the hypothesis that supplemental seminal plasma in extended ram semen improves conservation at 5°C, we added various concentrations of seminal plasma to semen during storage, and investigated subsequent sperm function in vitro. Semen was divided into three aliquots, extended in a commercial diluent (Triladyl) supplemented with 0, 10 or 25% (vol:vol) ovine seminal plasma and cooled to 5°C. After 8 and 24 h at 5°C, sperm were suspended in a modified synthetic oviduct fluid (SOF-m) at 39°C to mimic the female genital tract at insemination. Sperm aliquots were assessed for motility and chlortetracycline fluorescence after 0, 4 and 8 h in the SOFm. No significant differences were observed due to seminal plasma supplementation during conservation at 5°C or incubation in SOF-m at 39°C. However, decreased sperm motility and fewer non-capacitated sperm were observed concomitant with an augmentation of capacitated and acrosome-reacted cells during incubation in SOF-m. Therefore, the hypothesis that diluent supplementation with homologous seminal plasma improves ram sperm conservation or subsequent sperm function was not supported. Key words: Ovine, ram, sperm, motility, viability, chlortetracycline fluorescence, artificial insemination, SOF

Effect of sperm coating on the survival and penetrating ability of in vitro stored bovine spermatozoa

The aim of this study was to examine the effect of sperm coating on the survival and penetrating ability of in vitro stored diluted spermatozoa. Bovine semen was collected by means of an artificial vagina connected with a tube containing 5 ml of the commercial Triladyl diluent supplemented with 20% egg yolk and 6.7% glycerol (EYTG). Both EYTG and seminal plasma were removed by centrifugation and the spermatozoa were stored under different in vitro storage conditions. In the first and second experiment, “control” and “coated” spermatozoa were stored in Hepes-TALP (pH 6 and 7) at room temperature. After 4 days of storage, the progressive motility, membrane integrity, mitochondrial membrane potential or DNA integrity of the spermatozoa were evaluated before and after Percoll centrifugation. The in vitro penetration rate of the spermatozoa was examined only after Percoll centrifugation. A significantly ( P<0.05) positive influence of sperm coating was observed on the tested sperm characteristics and penetration rate of spermatozoa when they were stored in Hepes-TALP at pH 7, but not at pH 6. In the last experiment, the influence of the storage medium Hepes-TALP (pH 7) or EYTG was investigated on motility, membrane integrity, mitochondrial membrane potential and in vitro penetration potential of “coated” spermatozoa stored at room temperature or at 4 °C during 4, 5 and 6 days. After 6 days of storage, a significantly ( P<0.05) higher percentage of motile and membrane intact spermatozoa with high mitochondrial membrane potential was obtained in EYTG at both temperatures leading to a significantly higher in vitro penetration rate. These results indicate that sperm coating could preserve sperm characteristics and penetrating capacity of fresh bovine spermatozoa stored in egg yolk containing diluent for up to 6 days.