Reduced glutathione content in human sperm is decreased after cryopreservation: Effect of the addition of reduced glutathione to the freezing and thawing extenders.

In this study, total glutathione content was determined in human spermatozoa before and after cryopreservation. Total GSH in fresh semen was 4.47±0.46nmol/10(8) cells. Following semen cryopreservation, GSH decreased to 1.62±0.13nmol/10(8) cells, a 64% reduction (p<0.01). This decrease in GSH content was associated with a decrease in sperm progressive motility (68% of reduction, p<0.01). Addition of 1mM GSH to the freezing extender increased the percentage of total motility and sperm viability. It also modified the motility pattern measured by CASA with changes in the straight-line and average path velocities and wobble of the curvilinear trajectory. Addition of GSH to the freezing media reduced spermatozoa ROS levels and increased the level of sulfhydryl groups on membrane proteins. Nevertheless, no effect of GSH addition on lipid membrane disorder or chromatin condensation was detected. Addition of 1 or 5mM GSH to the thawing media increased the percentage of motile and progressively motile spermatozoa, but no effect on viability was detected. In conclusion, the antioxidant defensive capacity of the GSH is severely altered by the freeze-thawing process. The addition of GSH to the freezing and thawing extender could be of partial and limited benefit in improving the function of frozen human spermatozoa.

Clinical and laboratorial description of the differential diagnoses of hemostatic disorders in the horse.

The process of fibrin clot formation is a series of complex and well-regulated reactions involving blood vessels, platelets, procoagulant plasma proteins, natural inhibitors, and fibrinolytic enzymes. Vasculitis can be caused by a variety of different agents as bacteria, viruses, protozoal, rickettsial organisms, toxic, drugs, medications, and neoplasms. The most common cause of vasculitis is the purpura hemorrhagica, which is associated with exposure to Streptococcus equi ssp. equi or less commonly, equine influenza. Deficiencies or defects of the hemostatic components may result in bleeding and/or thrombosis. Inherited alterations of primary hemostasis (von Willebrand disease: vWD and Glanzmann's thrombasthenia: GT) and of secondary hemostasis (hemophilia A and prekallikrein: PK deficiency) are scarcely reported in equine clinic. On the contrary, acquired alterations of primary and secondary hemostasis are commonly found. They include thrombocytopenia, platelet dysfunction due to the administration of some drugs and targeted antiplatelet agents, decreased factor synthesis (liver disease or deficiency of vitamin K), release of inactive factors, inhibition of factor activity, or excessive consumption and depletion of factors (platelets, coagulation factors, and anticoagulants factors as antithrombin (AT) and protein C). Disseminated intravascular coagulation (DIC) is the most common and complex hemostatic disorder in horses and appears to be associated with sepsis, inflammatory and ischemic gastrointestinal tract disorders and other systemic severe diseases. These alterations are commonly found in patients in intensive care units.

Anthropic impacts on Sub-Saharan urban water resources through their pharmaceutical contamination (Yaoundé, Center Region, Cameroon).

Sub-Saharan urban centers have to tackle high population growth, lack of sanitation infrastructures and the need for good quality water resources. To characterize the impacts of anthropization on the water resources of the capital of Cameroon (Yaoundé), a multi-disciplinary approach was used in ten sub-watersheds (peri-urban and urban) of the Méfou watershed. Pharmaceutical residues were used as tracers of surface and groundwater contamination caused by the release of domestic wastewater from pit latrines and landfills. A water use survey was conducted in the vicinity of the sampling sites to better assess water use, treatment and management. Available land use and hydro-geomorphological data completed characterization of the sub-watersheds. The combined data showed that natural features (elevation, slope, and hydrography) and human activities (land use) favor rainfall-runoff events and hence surface water contamination. Pharmaceutical monitoring revealed contamination of both surface and groundwater especially in the urban sub-watersheds. Analgesics/anti-inflammatory drugs and anti-epileptic carbamazepine were the most frequently found compounds (in up to 91% of water samples) with concentrations of acetaminophen reaching 5660 ng/L. In urban sub-watersheds, 50% of the groundwater sites used for drinking water were contaminated by diclofenac (476-518 ng/L), carbamazepine (263-335 ng/L), ibuprofen (141-276 ng/L), sulfamethoxazole (<2-1285 ng/L) and acetaminophen (110-111 ng/L), emphasizing the need for a deeper understanding of the interactions between surface and groundwater. The use of groundwater as drinking water by 68% of the total population surveyed raises concerns about population exposure and potential health risks. This case study highlights the need for strategies to limit contamination of the water resource given the predicted future expansion of Sub-Saharan urban centers.

Effect of open pulled straw (OPS) vitrification on the fertilisation rate and developmental competence of porcine oocytes.

Freezing technologies are very important to preserve gametes and embryos of animals with a good pedigree or those having high genetic value. The aim of this work was to compare immature and in vitro matured porcine oocytes regarding their morphology and ability to be fertilised after vitrification by the open pulled straw (OPS) method. In four experiments 830 oocytes were examined. To investigate the effect of cumulus cells on oocyte survival after OPS vitrification, both denuded and cumulus-enclosed oocytes were vitrified at the germinal vesicle (GV) stage, then after vitrification they were matured in vitro. Besides, in vitro matured oocytes surrounded with a cumulus and those without a cumulus were also vitrified. The survival of oocytes was evaluated by their morphology. After in vitro fertilisation the rates of oocytes penetrated by spermatozoa were compared. Our results suggest that the vitrification/warming procedure is the most effective in cumulus-enclosed oocytes (22.35 +/- 1.75%). There was no difference between the order of maturation and vitrification in cumulus-enclosed oocytes, which suggests the importance of cumulus cells in protecting the viability of oocytes during cryopreservation.

The effect of nerve growth factor on nuclear progression of porcine oocytes during in vitro maturation and embryo development.

The present study examined the effect of nerve growth factor (NGF) on in vitro maturation (IVM), in vitro fertilisation (IVF) and subsequent embryonic development of porcine oocytes. Cumulus-oocyte complexes were cultured with or without 1.0 ng/ml NGF for 40 h. After IVF, they were cultured in vitro for 6 days. After 10 and 20 h of IVM, there was no difference in nuclear status between the NGF-treated and control oocytes. Significant differences were detected in nuclear progression of oocytes matured in the presence or absence of NGF at 30 h of culture. A higher proportion of NGF-treated oocytes were at M-II stage compared to the control. Nevertheless, at the end of the 40-h IVM period, there was no difference in the proportion of M-II stage oocytes between the NGF-treated and control groups. NGF in IVM medium did not influence the developmental competence of putative embryos. Most embryos remained at the 2- to 4-cell stage; however, a significant amount of embryos reached the morula stage both in the NGF and the control groups. These results suggest that NGF during IVM accelerates nuclear progression of porcine oocytes by enhancing the post-diakinetic events of meiosis.

Three probable cases of Loa loa encephalopathy following ivermectin treatment for onchocerciasis.

Over the past nine years, more than 12 million people exposed to Onchocerca volvulus infection have received at least one dose of ivermectin, almost all without serious adverse reactions. Since 1991, however, several cases with neurologic manifestations, including coma, have been reported after ivermectin treatment of persons infected with O. volvulus who also had concomitant Loa loa infection with very high microfilaremia (> 50,000 microfilariae/ml of blood). In 1995, four criteria were established to define probable cases of Loa encephalopathy temporally related to treatment with ivermectin (PLERI). The present paper describes three PLERI cases recorded in Cameroon and compares them with two others reported previously. Disorders of consciousness began 3-4 days after treatment. The objective neurologic signs were variable. The conditions improved favorably in three patients who benefited from early hospitalization and good nursing; their disorders of consciousness lasted only 2-3 days; the results of clinical examination became normal after one month and electroencephalographic abnormalities disappeared after 5-7 months. Conversely, late diagnosis and delay in proper management in two others probably led to worsening of the condition and to fatal outcome related to the usual complications of coma. In addition to these cases, patients w with high Loa microfilaremia also developed milder neurologic manifestations causing functional impairment lasting for at least one week after treatment. Before launching mass ivermectin distribution programs to control onchocerciasis in central Africa, communities in which the intensity of concomitant L. loa microfilaremia is high need to be identified, and specific educational measures and monitoring strategies should be developed and applied before they are treated.

Tonate virus infection in French Guiana: clinical aspects and seroepidemiologic study.

Two recent cases of human infection with Tonate virus, one of which was a fatal case of encephalitis, have renewed interest in these viruses in French Guiana. The clinical aspects of confirmed and probable cases of infection with this virus indicate that it has pathogenic properties in humans similar to those of other viruses of the Venezuelan equine encephalitis complex. To determine the prevalence of antibodies to Tonate virus in the various ethnic groups and areas of French Guiana, 3,516 human sera were tested with a hemagglutination inhibition test. Of these, 11.9% were positive for the virus, but significant differences in seroprevalence were found by age, with an increase with age. After adjustment for age, significant differences were found between places of residence. The prevalence of antibody to Tonate virus was higher in savannah areas, especially in the Bas Maroni (odds ratio [OR] = 22.2, 95% confidence interval [CI] = 15.2-32.4) and Bas Oyapock areas (OR = 13.4; 95% CI = 9.8-18.4). The ethnic differences observed in this study were due mainly to differences in place of residence, except that whites were significantly less frequently infected than other ethnic groups. This study indicates that Tonate virus infection is highly prevalent in French Guiana, especially in savannah areas.

Ivermectin-based control of onchocerciasis in northern Cameroon: individual factors influencing participation in community treatment.

A study aimed at determining individual factors associated with participation in community treatment with ivermectin was conducted in a village hyperendemic for onchocerciasis in northern Cameroon. The respective influences of sex, age, place of residence, distance between the compound and the dosing point, compound size, and participation in treatment by authoritative individuals in the compound was evaluated using univariate and multivariate analysis. Participation in treatment was closely associated with the attitude of the compound heads. Participation of compound heads in treatment increased as the household size increased, and as the distance to the distribution point diminished. This may be explained by the fact that getting information on health programmes is easier in large households whose members are involved in various social activities, and in compounds located near the village centre. Staff involved in health education should take this issue into account, and try to ensure circulation of information particularly to those living in small or remote compounds.

Efficacy of repeated doses of ivermectin against Mansonella perstans.

Treatment of Mansonella perstans infection, although seldom necessary, is difficult. In a 3 year's trial of normal and high-dose annual and 3-monthly ivermectin treatment against Onchocerca volvulus, the effects on M. perstans were recorded and related to the cumulative dose received. The World Health Organization's African Programme for Onchocerciasis Control may thus reduce the endemicity of M. perstans.

Ocular findings after ivermectin treatment of patients with high Loa loa microfilaremia.

Hemorrhages in the palpebral conjunctiva (HPCs) have been recorded in patients living in an area endemic for loiasis who developed serious reactions after ivermectin treatment. A study was designed to evaluate the frequency of these lesions, and to identify risk factors associated with their appearance. The conjunctivae of 1,682 patients who complained of reactions were systematically examined. HPCs were found in 41 patients. The initial mean Loa loa microfilaremia in the individuals with HPCs was 14,900 microfilariae (mf) per mL, as compared with 14.5 mf/mL in the other patients. Mansonella perstans microfilaremia and male gender were also associated with HPCs. Post-treatment fundus examinations were performed on 37 patients, and a close relationship was found between the occurrence of HPCs and the presence of retinal lesions. The vascular pathological processes leading to the ocular lesions may be similar to those which occur at the cerebral level in patients harboring high L. loa microfilaremia who develop neurologic troubles after ivermectin treatment. Retinal lesions may represent a special feature of the Loa-related encephalopathies useful for differential diagnosis, and the HPCs may be useful as an alarm sign to identify those individuals who might develop serious reactions after ivermectin treatment.

Successful intercontinental transport of porcine embryos from Europe to South America.

A total of 393 porcine embryos was transported in two trials each lasting more than 30h from Hannover, Federal Republic of Germany to Buenos Aires, Argentina. The embryos were shipped in phosphate buffered saline (PBS) medium supplemented with 10% lamb serum and packaged in 0.25ml straws placed in a modified thermos bottle at 36.5 degrees C. Upon arrival, 359 embryos (90.8%) were evaluated as morphologically intact and were transferred to 19 recipients. Twelve recipients remained pregnant. Three recipients aborted and nine recipients (47.4%) farrowed a mean number of 5.6 +/- 2.6 (x +/- SD) piglets after 115.1 +/- 1.8 d of gestation. The average birthweight was 1.1 +/- 0.2 kg. The percentage of embryos resulting in live piglets was 28.6% in farrowing recipients. These births represented the first piglets from embryos that had been stored for more than 30h in vitro and shipped from Europe to South America.

Sexing in vitro produced bovine embryos, at different stages of development, using rat H-Y antiserum.

The male-specific H-Y antigen is present on mammalian cell membranes and has been identified by various methods, including antiserum cytotoxicity. The objective of the present study was to determine the sex of in vitro produced (IVP) bovine embryos, at varying stages of development, by culturing in the presence of rat monoclonal H-Y antibodies. Embryos derived from IVM/IVF were classified according to the interval after IVF (48, 96 or 120 h) as Category 1, 2 or 3 if they had 4 to 8, <32, and >32 cells, respectively. Embryos of each category were cultured for 24h in TCM-199 supplemented with bovine oviductal epithelial cells, fetal calf serum (FCS), and antibiotics (Control group), to which the following had been added: guinea pig serum (GPS; C' group); H-Y antiserum (HY group); or GPS and H-Y antiserum (C' + HY group). After culture, embryos were designated as "affected" when development was arrested or one or more blastomeres was degenerate; embryos lacking these changes were designated "unaffected." The sex of each embryo was subsequently determined by chromosome analysis. After 48h of IVF (Category 1), within each of the four treatments, the proportion of unaffected embryos was higher than the proportion of unaffected embryos (81% versus 19%, P < 0.05). Similarly, the Control, C' and HY groups of Categories 2 and 3 embryos had different proportions of unaffected versus affected embryos (75% versus 25%, P < 0.05). In all these groups, the male:female ratio did not significantly differ from 1:1. In contrast, in the C' + HY group of Categories 2 and 3 embryos, the ratio of unaffected versus affected embryos was 41% versus 59% (P < 0.05) and the male:female ratio differed (P < 0.05) from the expected 1:1 ratio (approximately 0.3:1 and 4.5:1 for unaffected versus affected, respectively). In conclusion, when bovine embryos were cultured in the presence of rat monoclonal H-Y antibodies and compliment, alterations occurred in embryos that were beyond the 8-cell stage; we inferred that the antibodies cross-reacted with H-Y antigens.

[Comparison between various methods of pregnancy screening during a large-scale ivermectin treatment in Cameroon]. / Comparaison entre différentes méthodes de dépistage des grossesses au cours de traitements par ivermectine à large échelle au Cameroun.

During a mass treatment with ivermectin which is contraindicated to pregnant women, authors made a screening for pregnancy by questioning 2,580 women from 15 to 45 years old. 1,409 of these women were also interrogated by a female physician assisted by a native matron to detect pregnant women. In 1,798 women, at least one immunological pregnancy test has been used in the field. A nine months follow-up was made to check answers. Women's knowledge led to a good appreciation of pregnancy as they correctly appreciated their pregnancy from the second month: specificity was good (98%) but sensitivity was moderate (71%). Conclusions made by gynecological team after detailed questionnaire were more sensitive (80%), but specificity decrease dramatically to 59%. Combinations of several items in the questionnaire showed no benefit. Sensitivity of the various immunological pregnancy tests tried was included in 70 to 90% and specificity was varying from 87 to 97%. Simple questioning of women was the most efficient method (efficiency value was 94% against 63% for interview and 92% for pregnancy tests). However, the use of this method induced the risk that about 29% of women have been treated although they were pregnant.

[Epidemiological features of retroviral infection by HTLV-1 in central Africa]. / Particularités épidémiologiques de l'infection rétrovirale a HTLV-1 en Afrique centrale.

Subsaharian Africa is the most important reservoir of HTLV1 virus but its epidemiology is not well-known. The authors have studied from 1987 to 1990 the situation in 6 countries in Central Africa. The already described routes of transmission are present: through blood transfusions (but it's not such a common practice), heterosexual transmission (but it doesn't seem to be as important as in other continents), and mother to child transmission through breast feeding. Nevertheless, environmental factors--which have to investigated--could play a role for transmission, among them one could think to vectors and parasites.

[Secondary effects of the treatment of hypermicrofilaremic loiasis using ivermectin]. / Effets secondaires du traitement de la loase hypermicrofilarémique par l'ivermectine.

In the last ten years ivermectin appeared an efficient and safe alternative to diethylcarbamazine which is known to induce severe adverse reactions in loiasis, including encephalitis. After these results, large scale ivermectin treatments against onchocerciasis were carried out in Central Africa where loiasis is also endemic; and seven cases of severe reaction were reported in Cameroon since 1991, during these mass ivermectin treatments. In order to study adverse reactions in patients harbouring high load of Loa loa microfilariae (mf), we realized careful hospital based treatment in 112 patients with more than 3,000 mf/ml (ml) blood. Patients received once 200 micrograms ivermectin per kilogram at day 0 (D0). Clinical examination was made daily during the four following days (D1 to D4). Blood and urine samples were analysed before treatment and at D1 and D3. Lumbar puncture was made at D1 for 39 patients with more than 10,000 mf/ml; at D3 for the 49 following patients without consideration for the level of parasitaemia, and at D0 and D3 for ten voluntary patients. For analysis the patients were distributed in 3 groups according to initial parasitaemia: the first group included 50% out of the patients, those whose parasitaemia was fewer than 15,000 mf/ml blood; the second group included 25% patients whose parasitaemia was between 15,000 and 30,000 mf/ml; the third group included the last 25% patients whose parasitaemia was higher than 30,000 mf per ml blood. Adverse reactions were observed in 71% out of the patients. Symptoms described were fever, pruritus, headache, arthralgia. Most symptoms appeared 24 to 36 hours after treatment. Temperature increased significantly in group 3. Microfilaraemia decreased by 85% in the 3 groups during the 4 days following treatment. C-reactive-protein increased dramatically after treatment in all patients (p < 10(-4)). Some patients presented blood in urine in three groups but haematuria reached 35% of patients in group 3. Proteinuria is noted among 33% of all patients but 20% in group 1 and 2 versus 70% in group 3. Loa loa mf were observed in urine of half the patients, but in low amounts (< 10 mf per 50 ml urine). In cerebro-spinal fluid (CSF), some mf appeared at D1 or D3 in people heavily infected with Loa loa, reaching 80% of the patients of group 3. LP made at D0 in ten patients with parasitaemia higher than 30,000 mf/ml blood confirmed that CSF was naturally microfilaria free before treatment. One patient presented severe troubles with fever, asthenia and conscience troubles beginning at D3, reactive coma at D4, renal impairment with transitory anuria; progressive improvement in 2 weeks and complete recovery at D22; he presented 102 mf/ml CSF at D6. The study confirmed that ivermectin treatment is generally well tolerated. Among people with high Loa loa parasitaemia the symptoms after treatment are frequent but mild. However severe cases with conscience troubles are possible, and may occur in about 1% of subjects with more than 3,000 mf/ml blood. Severity of adverse reactions was linked to level of parasitaemia before treatment. The critical parasitaemia level which could lead to expect serious adverse effects seems to be 30,000 ml/ml blood. These informations should induce carefulness to carry out large scale treatments against filariosis in endemic areas of Loa loa.

[Analysis of the prevention of post-ivermectin Loa loa encephalopathy by administration of initial low dose]. / Essai de prévention des encéphalopathies à Loa loa post-ivermectine par l'administration d'une faible dose initiale.

A number of cases of Loa-encephalopathy have been reported following ivermectin (Mectizan) treatment for onchocerciasis in patients with high Loa microfilaraemia. A possible explanation for these severe reactions is the formation of micro-emboli in small brain vessels as a result of massive paralysis of Loa microfilariae in the blood. This suggests that encephalopathy might be prevented by giving an initial low dose of ivermectin to induce a more gradual action on the Loa microfilariae. To test this hypothesis, a trial was conducted on 23 adult patients in Cameroon. Patients were divided into two groups. One group received the recommended dose of 150 micrograms/Kg. The other group received a lower dose of 50 micrograms/Kg (one 3 mg tablet of Mectizan). Blood smears were made daily from day 1 to 7 after treatment and then on days 15 and 30. Results showed no difference in the effect of the dosage level on Loa microfilaraemia. This finding suggests that an initial low dose of 3 mg Mectizan will not prevent encephalopathy following treatment for onchocerciasis.

[AIDS and health personnel in Ivory Coast]. / SIDA et personnel soignant en Côte d'Ivoire.

During the second half of 1993, a survey to assess awareness, attitudes, habits, and behavior with regard to HIV and AIDS was performed among health care workers in the Cote d'Ivoire. A total of 90 health care workers, 42 in Abidjan and 48 in Daloa, were surveyed. The majority (87%) were women. Understanding of the terms "AIDS" and "seropositivity" was good as was knowledge of AIDS-related symptoms. Awareness was better among newly recruited workers and nurses than among orderlies. Appreciation of risk was strongly linked with the notion of high-risk population. The occupational nature of the risk was clearly understood and anxiety in this regard was increased by uncertainty as to modes of secondary transmission and lack of preventive measures. On the individual and personal level protection consisted mainly in reducing the number of partners and outside activities. There was a great demand for training and information. As a result of the uncertainties revealed in this survey there appears to be a reluctance or refusal to perform certain procedures or treat certain patients and even a tendency to abandon the health care profession.