Egg yolk can be partially replaced with carboxylated poly-l-lysine (cpll) in extender for nili- ravi buffalo bull sperm.

BACKGROUND: Egg yolk is inevitably associated with risks of microbial contamination and anti-cryoprotectant agents that necessitate the investigation of some synthetic alternatives. OBJECTIVE: To investigate the potential of carboxylated poly-L-lysine (CPLL) as a replacement for egg yolk during the cryosurvivability of Nili-Ravi buffalo sperm. MATERIALS AND METHODS: Semen collected from four Nili-Ravi buffalo bulls (two ejaculates / bull / day; total 40 ejaculates for five replicates) was cryopreserved in different experimental extenders viz: Control (CPLL 0%, egg yolk 20%); E1 (CPLL 5%, egg yolk 15%); E2 (CPLL 10%, egg yolk 10%); E3 (CPLL 15%, egg yolk 5%) and E4 (CPLL 20%, egg yolk 0%). Post-thaw quality was assessed in terms of sperm motility, plasma membrane integrity (PMI), viability, live:dead ratio, lipid peroxidation of sperm and total antioxidant capacity of seminal plasma. RESULTS: Sperm motility improved (P<0.05) in extenders replacing 5%, 10% and 15% egg yolk with CPLL. Sperm PMI, viability and live:dead ratio also improved (P<0.05) in extenders replacing 10%, 15% and whole (20%) egg yolk with CPLL. In contrast, sperm DNA integrity was not different (P>0.05) when CPLL replaced egg yolk at any level. The lipid peroxidation level decreased with a concomitant increase in total antioxidant activity of seminal plasma when CPLL replaced egg yolk at 5%, 10%, 15% and 20%. CONCLUSION: Replacement of 15% egg yolk in the extender with CPLL improves all sperm quality parameters: motility, PMI, viability, live:dead ratio, lipid peroxidation of sperm and total antioxidant activity of seminal plasma.

Antioxidant activity of Nigella sativa Seeds Aqueous Extract and its use for cryopreservation of buffalo spermatozoa.

The free radical scavenging activity (RSA) of Nigella sativa extract and its efficiency for cryopreservation of buffalo spermatozoa was investigated. In experiment 1, Nigella sativa extract was prepared and evaluated for RSA using 2,2-diphenyl-1-picrylhydrazyl assay. The results showed increased pattern of RSA at 1%-5% of Nigella sativa extract. In experiment 2, buffalo semen from three bulls (24 ejaculates) was incubated at 0%, 0.1%, 0.3%, 0.5%, 1%, 1.5%, 2%, 3%, 4%, 5% and 6% extract to assess in vitro tolerability to Nigella sativa in terms of progressive motility (PM). Buffalo spermatozoa showed tolerance to all levels; rather, sperm PM was increased at 1%-4% extract. In experiment 3, semen from three bulls (24 ejaculates) was cryopreserved with 0%, 1%, 2%, 3%, 4% and 5% of Nigella sativa extract. Sperm PM and plasma membrane integrity (PMI) were evaluated after dilution and cooling, while PM, PMI, viability and DNA integrity were evaluated after thawing. Nigella sativa extract at 4% in extender improved (p < .05) post-dilution, post-cooling and post-thaw sperm quality. In conclusion, Nigella sativa extract at all concentrations (1%-6%) showed antioxidant activity and its supplementation at 4% in extender improved buffalo sperm quality at all stages of cryopreservation.

Sperm preparation through Sephadex™ filtration improves in vitro fertilization rate of buffalo oocytes.

Routinely, swim-up method is used to separate high-quality sperm; however, long processing time and close cell-to-cell contact during the centrifugation step are inevitable elements of oxidative stress to sperm. The objective was to evaluate Sephadex™ and glass wool filtration to separate motile, intact and viable sperm for in vitro fertilization in buffalo. The cumulus-oocyte complexes (COCs) were collected from ovaries of slaughtered buffaloes by aspiration and matured for 24 hr in CO2 incubator at 38.5°C and 5% CO2 . Matured COCs were rinsed twice in fertilization TALP and placed in the pre-warmed fertilization medium without sperm. Cryopreserved buffalo semen was thawed at 37°C for 30 s and processed through Sephadex™ , glass wool filtration and swim-up (control). Total and motile sperm recovery rates were assessed, resuspended in fertilization TALP and incubated for 15-20 min in CO2 incubator. Samples prepared by each method were divided into two aliquots: one aliquot was studied for sperm quality (progressive motility, membrane integrity, viability, liveability), while the other was subjected to co-incubation with sets of 10-15 in vitro matured oocytes. Data on sperm quality were analysed by ANOVA, while in vitro fertilizing rates were compared by chi-squared test using SPSS-20. Least significant difference (LSD) test was used to compare treatment means. Glass wool filtration yielded higher total and motile sperm recovery rate, while Sephadex™ filtration improved (p < .05) sperm quality (progressive motility, membrane integrity, viability, liveability). Sperm preparation through Sephadex filtration yielded higher in vitro fertilization rate in terms of cleavage rate compared to glass wool filtration and swim-up (control). In conclusion, cryopreserved Nili-Ravi buffalo sperm selected through Sephadex filtration showed improved quality and yielded better fertilization rates (cleavage rate) of in vitro matured/fertilized oocytes. Sephadex filtration could be a promising technique for use in in vitro fertilization in buffalo.

Oxygen use for chest pain in coronary care units across the UK.

AIM: To quantify the adherence to national guidance for the use of oxygen in patients presenting with chest pain to coronary care units (CCUs) across the UK. DESIGN: Prospective survey. METHODS: A total of 307 hospitals were contacted by telephone between August 2010 and October 2010. Of these, 48 had no CCUs, 10 units refused to take part and 18 hospitals were contacted on 2 occasions but were unable to provide the information due to paucity of time owing to heavy clinical workload. Overall 231 hospitals participated in the audit questionnaire. RESULTS: A total of 30% of the units used oxygen titrated to saturations in accordance with national guidelines. There was no difference between units that had on-site availability of percutaneous coronary intervention and those that did not. Those hospitals where there was a policy for routine oxygen prescription were as unlikely to comply with the guidelines on oxygen use as hospitals where oxygen was not routinely prescribed. CONCLUSION: Only one-third of CCUs in the UK reported adherence to guidelines with regards to oxygen delivery in patients presenting with chest pain. Despite this figure seeming rather low, this is consistent with practice through a range of specialties and guidelines. The evidence base for the oxygen guidance remains insecure. Additional research is required but in the meantime we recommend oxygen is prescribed according to current guidelines.

Surgical outcome and risk stratification for primary retinal detachment repair: results from the Scottish Retinal Detachment study.

OBJECTIVES: To report the early surgical outcome, risk of failure and predictive value of rhegmatogenous retinal detachment (RRD) classification based on all participants in the Scottish Retinal Detachment study. METHODS: Over 2 years, all incident cases of RRD in Scotland were approached for recruitment. Early postoperative success was defined as an attached retina following one procedure with a minimum follow-up of 6-8 weeks. Using a regression model, the influence of clinical factors on the failure risk was estimated and the sensitivity and specificity of the Royal College of Ophthalmologists (RCOphth) grading for RRD and the vitrectomy in retinal detachment stratification risk formula (VR-SRF) in predicting operative failure were assessed. RESULTS: Primary outcome data were available for 86.2% (975/1130) of patients. The overall primary success rate was 80.8% (95% CI 78.1 to 83.3%). The presence of preoperative proliferative vitreoretinopathy of any degree and each additional clock hour of detachment increased the risk of failure by an OR of 2.4 and 1.13 respectively (p<0.05). A specificity of >95% in predicting early surgical failure was noted for highly complex RRDs according to the VR-SRF formula and the RCOphth classification. CONCLUSIONS: Consistent with previous series, the overall early success rate of RRD repair was 80% after one operation. The type of surgical repair did not influence overall success rates. Significant predictors of failure are the presence of preoperative proliferative vitreoretinopathy of any grade and the extent of detachment. The analytical value of current classification systems in predicting failure is most useful in complex RRDs.

Pilot randomised controlled trial of face-down posturing following phacovitrectomy for macular hole.

BACKGROUND: To gather information on the effect of postoperative face-down posturing following combined phacoemulsification and vitrectomy for macular hole surgery in order to assist in the design of a larger definitive study. METHODS: Thirty phakic patients with stage II-IV full-thickness macular hole had combined phacoemulsification and pars plana vitrectomy with internal limiting membrane peel and 14% perfluoropropane (C(3)F(8)) gas. At the conclusion of surgery, patients were randomised either to face-down posture or to no posture, for 10 days. The primary outcome was macular hole closure. RESULTS: The macular hole was successfully closed in 93.8% of the face-down posture group and in all of the no-posture group. Mean visual improvement was 0.63 (SD=0.21) logMAR units in the face-down group and 0.53 (SD=0.22) in the no posture patients. CONCLUSION: Following combined phacoemulsification and vitrectomy, postoperative face-down posturing appears to make little difference to the final anatomical or visual outcome. If we assume a success rate of 95% in the posturing arm, and that there is no difference between posturing and non-posturing, then 798 patients would be required to be 90% sure that the 95% confidence interval will exclude a difference of more than 5%.

The outcome of deep anterior lamellar keratoplasty in herpes simplex virus-related corneal scarring, complications and graft survival.

AIMS: To determine the visual outcome, graft survival and complications after deep anterior lamellar keratoplasty (DALK) in patients with herpes simplex virus (HSV)-related corneal scarring. METHODS: A retrospective analysis of the patients who had DALK for HSV-related corneal scarring between January 2004 and February 2007 was performed. Mean follow-up was 30 months (range 16-48 months). The statistical significance of host corneal vascularisation was determined using Fisher's exact test. RESULTS: There were 18 eyes from 18 patients and the mean age was 57 years. Preoperative visual acuity ranged from hand movements (HM) to 6/12. Fifty per cent of the eyes achieved visual acuity of 6/12 or better postoperatively. Six eyes (33%) had recurrence of HSV-related inflammation, eight eyes (including four eyes with recurrence of HSV-related inflammation) developed graft rejection and four eyes (including two eyes with recurrence of HSV-related inflammation) had bacterial keratitis. The graft survival rate was 83%. Three eyes developed glaucoma and one eye required trabeculectomy. Immunohistochemistry revealed that HSV was focally positive or equivocal in four recipient corneal buttons, and transmission electron microscopy showed intracellular HSV virions in two of them. CONCLUSIONS: This is the largest series of DALK for herpetic corneal scarring that shows a comparable visual outcome and better graft survival rate than penetrating keratoplasty. There is significant risk of recurrence of HSV-related inflammation and graft rejection that requires timely recognition and adequate management.

Penetration of topical and subconjunctival corticosteroids into human aqueous humour and its therapeutic significance.

Topical and subconjunctival corticosteroids are some of the most effective and compelling treatment options in ocular inflammatory diseases. A systematic review of literature indexed by Ovid MEDLINE & EMBASE was performed up to December 2008. There are few studies on their aqueous penetration in human subjects. This review article discusses the penetration of different ocular corticosteroids into human aqueous humour along with the therapeutic implications on management of ocular surface diseases, immune-related corneal diseases, anterior uveitis and postoperative anti-inflammatory use. In the context of the paucity of well-constructed, prospective clinical trials comparing the efficacy of different corticosteroids, it provides guiding principles for the use of topical corticosteroids. Dexamethasone alcohol 0.1% and prednisolone acetate 1% are potent corticosteroids, but the latter achieves the highest aqueous concentration within 2 h and maintains higher levels for 24 h. Subconjunctival corticosteroids provide very high concentrations in the aqueous which maintain higher concentrations for longer periods.

Review of photodynamic therapy.

INTRODUCTION: Photodynamic Therapy (PDT) is an emerging treatment for a variety of conditions including ocular and extra ocular diseases. The porphyrins have been used extensively, as dyes, which are laser-activated to achieve desired clinical effects. Commonly used agents are verteporfin and porfimer sodium. METHODS: We performed a literature search of the PubMed database using the medical search headings: photodynamic therapy, photosensitizer verteporfin, visudyne, porfimer sodium and photofrin. We also performed a manual search using references from these articles, review articles and manufacturers' product monographs. RESULTS: Verteporfin and porfimer sodium are commonly used photosensitizing agents with their wide applications in different fields of medicine. Both have well established safety profiles. They are most commonly used in wet age-related macular degeneration, gastrointestinal diseases and bronchial cancers. CONCLUSION: PDT is a well established treatment entity in ophthalmology and other medical fields. In ophthalmology, it has rekindled interest and hopes in the common yet sight-threatening problem of age-related macular degeneration (AMD). This problem is still considered to be a serious issue as far as management is concerned. However in selective cases of AMD, it has shown success in restoring sight, especially in the 'classic' form of the disease. PDT is also being used to treat a range of solid cancers and non malignant conditions

Heterogeneity of staphylococci and other bacteria isolated from six-week-old broiler chickens.

In broiler operations, various health problems develop during the final 2 wk of the growing period, resulting in increased mortality and condemnation losses. At this stage, sickly birds were found to be systemically infected by various bacteria regardless of varied clinical signs, and the purpose of this study was to carry out thorough microbiological investigations on this problem. Thirty-one 6-wk-old broilers showing signs of illness were obtained from three farms, and bacterial isolations were carried out from the blood, liver, and hock joint. Bacteria were isolated from 87, 90, and 71% of the blood, liver, and hock joint samples, respectively. Mean bacterial counts in log10 of the blood (per milliliter) and liver (per gram) were 2.15 and 2.93, respectively. Among 132 bacterial isolates, major species were Staphylococcus (60%), Corynebacterium (18%), Escherichia coli (5%), and Stomatococcus (4%). Among 79 Staphylococcus isolates, 77 were coagulase-negative. Major species of staphylococci were S. lentus (19%), S. simulans (18%), S. cohnii (13%), S. gallinarum (10%), and S. captis (7%). In addition, six species of gram-positive and five species of gram-negative organisms were isolated. Thus, the apparent systemic infections were not caused by predominant pathogenic bacterial species, and adequately described as mixed infections. There were some significant relationships between isolated bacterial species and sampling sites, suggesting that certain organisms were abundant in the environment of a particular poultry house. These results indicate that systemic infections in market age broilers are caused by mixed bacterial species and suggest that they are caused by suppressed host antibacterial systems rather than pathogenic factors of microorganisms.

The epidemiology of Newcastle disease in rural poultry: a review.

Rural poultry are the dominant form of poultry kept in the developing world. They are a natural resource whose potential is not fully exploited for the welfare of rural populations. The most devastating disease of rural poultry is Newcastle disease (ND). ND is an epidemic disease in intensive poultry and is responsible for high economic losses. Its epidemiology in intensive poultry is well understood, but little has been published on its behaviour in village poultry populations. Some research has been carried out during the past few years and it now appears that rural poultry are receiving increasing support for research and development from many government and international funding agencies throughout the world. All strains of Newcastle disease virus (NDV) occur in rural poultry, but velogenic strains are reported to be more common. Serological surveys in conjunction with isolation studies have shown that velogenic NDV strains are endemic in rural poultry populations even in isolated villages and possibly in isolated flocks. Although NDV is endemic in village poultry, the clinical disease usually follows an epidemic pattern. ND outbreaks often occur once or twice a year at regular intervals affirming the endemicity of the virus, however, 'mini' outbreaks in individual flocks and sporadic cases in individual birds may occur. Epidemics usually occur at times of climatic stress, leading to seasonal occurrence. The spread of NDV within and between village poultry populations is relatively slow due to a low contact rate. The major mode of transmission appears to be by the faecal-oral route. The respiratory route may also play a role in flocks where close bird-to-bird associations exist. Other poultry species, wild and feral birds, wild animals, communal water reservoirs and domestic animals may play a role in transmission; however, their role has not been properly investigated. In non-immune, intensively reared poultry, introduction of a sufficient quantity of virulent NDV is sufficient to cause severe outbreaks, but in rural poultry epidemic disease depends upon a number of factors. These include NDV pathotype, age structure and immunity of the host population, breed susceptibility, concurrent disease and seasonal influences. Although ND is endemic in rural poultry, many aspects of the epidemiology and economics of ND have not been fully understood. Well-designed, longitudinal studies made within a socio-economic framework are required to improve our understanding of the main problems of rural poultry systems.

Oestrus ovis myiasis in Libyan sheep and goats.

The heads of 1,489 sheep and 320 goats were examined for larvae of Oestrus ovis at 17 abattoirs in northern Libya in July to November 1988. The prevalence of O. ovis in sheep was 22.6% and in goats it was 18.4 per cent. Up to 14 and 11 larvae were collected from individual sheep and goats respectively. All larvae were recovered from the nasal passages and frontal sinuses, but only second and third instars were seen.

Lice of farm animals in Libya.

This first report on the lice of farm animals in Libya lists a total of fifteen species, as follows: Haematopinus quadripertusus and Linognathus vituli from cattle, Bovicola ovis, Linognathus pedalis, L. africanus and L. ovillus from sheep, L. stenopsis, L. africanus and Bovicola caprae from goats. Poultry harboured Lipeurus caponis, L. heterographus, Menacanthus stramineus, Menopon gallinae, Goniocotes gallinae, Goniodes gigas and G. dissimilis.

A survey of mites on farm animals in Libya.

In 1985-1988, 2287 farm animals (cattle, camels, sheep, goats, horses, donkeys, dogs and rabbits) suspected of carrying parasitic mites were examined at 58 farms throughout Libya. Mites were identified on 1303 of these animals. The commonest parasites on cattle were Psoroptes and Chorioptes, on camels and sheep were Sarcoptes and Psoroptes, and on goats were Sarcoptes and Demodex. Infested horses carrier Psoroptes or Chorioptes, and one donkey carried Sarcoptes. Otodectes was common on dogs, but Sarcoptes was rare and no Demodex were seen. Rabbits often had psoroptic ear mange or sarcoptic body mange. Dermanyssus gallinae and Ornithonyssus bursa were seen on chickens, but no mites were found on pigeons, ducks or turkeys.

A survey of ticks on farm animals in Libya.

Thirteen species of ixodid ticks and two species of argasid ticks were collected during a three-year survey of 58 farms in Libya. These included Boophilus annulatus, B. microplus, B. decoloratus, seven species of Hyalomma, Rhipicephalus sanguineus, Rh. evertsi, Rh. bursa, Argas persicus and Ornithodoros foleyi. This is the first recording of B. microplus, B. decoloratus and Rh. bursa in Libya. Of 20,391 animals examined by random sampling, 2020 (9.6%) had ticks; particularly common were Hy. dromedarii on camels, Hy. impeltatum on sheep and Hy. excavatum on cattle. The tick found most frequently overall was Hy. dromedarii.

Experimental infection of Libyan sheep with Echinococcus granulosus.

The growth rate of unilocular hydatid cysts over a two-year period was studied in Libyan sheep dosed orally with gravid segments of Echinococcus granulosus taken from stray town dogs. Seven of the eight sheep dosed with 20-100 gravid segments (say 4000-20,000 eggs) had developed only infertile hydatid cysts after 107-611 days. Sheep dosed with 1000 gravid segments (say 200,000 eggs) had developed only sterile cysts when examined 358 days after dosing. Fertile cysts were recovered from the lungs after 428 days, and after 584 days from the liver. Only sterile cysts were seen in the heart, spleen and kidneys.