Cumulus Oophorus Expansion of Bovine Oocytes Cultured in vitro: A SEM and TEM Study

Bovine cumulus-oocyte complexes (COCs) were aspirated from antral follicles (3–6 mm in diameter). COCs with a compact multilayered cumulus investment were cultured for 9, 13 and 24 hours, respectively, and processed for scanning or transmission electron microscopy after different periods of culture including a 0 hour control group. In all 0 hour control COCs initial formation of cumulus cell projections, blebs and microvilli were observed. The start of cumulus-oocyte disconnection in cattle was observed after 9 hours of in vitro culture. These connections were almost completely discrupted after 13 hours, when a continuous production of extracellular matrix was observed. The full expansion of corona radiata cells was not observed even after 24 hours. Some cumulus oophorus cells were bound together with junctions of the zonula adherens type after 24 hours when extracellular matrix was found only in deeper layers. The full expansion of corona radiata cells was not the prerequisite for disconnection of cumulus-oocyte complex. Inhalt: Kumulus oophorus Expansion von in vitro kultivierten Rinderoozyten: Eine SEM und TEM Untersuchung Kumulus-Oozyten-Komplexe (COC) von Rindern wurden aus antralen Follikeln aspiriert (Durchmesser 3–6 mm). COC's mit kompakten mehrschichtigen Kumulus wurden für 9, 13 und 24 Stunden kultiviert und für die Scanner- oder Transmissionselektronenmikroskopie vorbereitet. Verschiedene Kulturzeiten incl. einer 0-Stundengruppe als Kontrolle wurden untersucht. In allen Kontroll COC's wurden beginnende Formationen yon Kumuluszellprojektionen, Bläschen und Mikrovilli beobachtet. Nach 9-stündiger in vitro Kultur begann die Lösung zwischen Kumulus und Oozyte. Die Verbindungen waren nach 13 Stunden vollständig unterbrochen, wobei eine kontinuierliche Produktion extrazellulärer Matrix beobachtet wurde. Eine vollständige Expansion der Corona radiata Zellen wurde auch nicht nach 24 Stunden beobachtet. Nach 24 Stunden waren einige Cumulus Oophorus Zellen über Verbindungen des Zonula adherens Typs verknüpft, wobei extrazelluläre Matrix nur in den tieferen Zellschichten gefunden wurde. Eine vollständige Kumulusexpansion ist keine Voraussetzung für die Auflösung des Kumulus-Oozyten-Komplexes.     

Fertilizing capacity of the ejaculate of nutria (Myocastor coypus) after the removal of the seminal vesicles as evaluated by the penetration test and natural mating

The fertility of male coypu sperm following seminal vesicle extirpation was investigated using the penetration test into the egg of Syrian golden hamster (Mesocricetus auratus). Ejaculates were obtained from five males by means of electro-ejaculation under halothane narcosis. The results of the zona-free hamster eggs (ZFHE) penetration test showed that the ejaculates of all the surgically treated coypu males were fertile and that ZFHE value fluctuated from 54 to 76.6%. The results obtained in experiments with natural mating revealed that the extirpation of male coypu seminal vesicles did not affect their fertility. In total 47 foetuses were found post mortem in ten coypu females covered by surgically treated males, which on average represented 4.7 foetuses per female.     

Model field study of Sumithion 50 EC and Fusilade S on pheasants.

Toxicological studies on wild animals play an important role in the ecotoxicological examination of pesticides. The applied model tests enable the assessment of toxicological consequences with particular regard to the life and nutrition of wild animals in the ploughed field among plants treated with pesticides. The application of different pesticide formulations on plough-land may pose a simultaneous chemical burden to wild birds. In this model study, manifestations of the interaction between an insecticide and a herbicide were studied in pheasants. The birds were placed on lucerne in cages (48 m2) and sprayed once. The applied doses were: Sumithion 50 EC 1 litre/ha + Fusilade S 6 litres/ha (practical doses) and Sumithion 50 EC 5 litres/ha + Fusilade S 30 litres/ha. The analytically determined pesticide concentration of the lucerne was taken as a basis in the further treatment of fodder. The fodder of pheasants contained the following chemicals: 85 mg/kg Sumithion 50 EC + 510 mg/kg Fusilade S and 425 mg/kg Sumithion 50 EC + 2250 mg/kg Fusilade S. Sporadic deaths observed among the pheasants were of traumatic origin and not due to a toxic effect. The decrease of body weight was significant only at the higher dose levels. Acetylcholinesterase (AChE) activity of the blood decreased significantly in both dose groups. On the basis of the results obtained it can be established that at the dose level used in the practice the pesticides studied do not give rise to a toxic interaction in pheasants.     

Chicken dendritic cells and type II pneumocytes express a common intracellular epitope

1. CVI-ChNL 74·3, a dendritic cell-specific monoclonal antibody (mAb) also identifies chicken lung granular pneumocytes (type II pneumocytes), which produce surfactant.

2. The 74·3 mAb does not cross-react with any other avian or mammalian granular pneumocyte, and provides a convenient tool for monitoring the status of type II pneumocytes in the chicken lung.     

Causes of anaphylactoid reactions in cattle after administration of lipoid preparations]

In 1986-1988, adverse anaphylactoid reactions (AR) were observed in animals in Czechoslovakia after the administration of oil adjuvant-containing vaccines or other lipoid drugs. Treated animals showed signs resembling the classic anaphylactic reaction, i.e. restlessness, salivation, pruritus, oedema and cyanosis of udder and vulva, and eyelid oedema, developing within a few minutes. The reactions were not elicited by the antigen alone, but by the oil adjuvant. The aim of our experiments was to identify substances eliciting the reaction in susceptible animals and to investigate possible induction mechanisms. The emulsifier Tween 80 has been demonstrated to be an AR inducing component of vaccines and drugs (Tab. I and III). Weak or moderate reactions were observed in 33% of animals treated with 5% Tween and 66% of those treated with 10% Tween showed strong reactions. On the other hand, no reactions were elicited by treatment with several paraffin oils of different quality (Tab. I) nor with an oil-in-water emulsion containing Montanid as an emulsifier (Tab. II). The role of the vegetative nervous system in the rise of AR has been confirmed. AR were suppressed in animals pretreated with parasympatholytic atropine and enhanced in a part of those pretreated with parasympathomimetic pilocarpine (Tab. III). The percentage of animals affected and the intensity of AR were also lower in animals pretreated with complement inhibitor epsilon-aminocapronic acid (Tab. IV). A major role of complement activation is suggested in the discussion of possible mechanisms of AR induction. It is possible to draw a conclusion on the basis of the results presented here and of the analysis of individual cases that a certain degree of animal susceptibility, depending on the phase of reproductive cycle, metabolism level and neurovegetative balance is necessary besides the administration of an AR inducer (Tween 80 in our case). Hence it seems that the adverse anaphylactoid reactions results from interactions of the two factors, i.e. administration of an AR inducer to susceptible animals.     

Murine typhus in the homeless

Homeless populations are particularly exposed to many vector-borne diseases because of their poor living conditions. We tested sera from 299 homeless people recruited in 2010 and 2011 in Marseilles, France for antibodies to Rickettsia typhi by microimmunofluorescence using a titer of 1:25 as a cut-off titer, and we confirmed the results by Western blot and cross-adsorption studies. Sixty-three persons (22%) had antibodies against R. typhi. The murine typhus seroprevalence rates have significantly increased in homeless populations between the 2000-2003 and 2010-2011 periods. These findings indicate that the homeless are increasingly exposed to flea-borne murine typhus in Marseilles. One might suggest that multiple strikes of sanitation workers resulting in the increase of waste and construction sites combined with the poor living conditions of the homeless expose this population to rodents and their fleas. Further annual studies are necessary to follow rodent-associated diseases among homeless people in Marseille.     

Radiodiagnostic examination of the swimbladder of some fish species

Radiodiagnostic methods have not been used previously for studying the anatomy and diseases of the swimbladder of freshwater fish species. In this study, the radiographic anatomy of the swimbladder and species-related differences in swimbladder structure were studied on plain radiographs taken of 12 Hungarian fish species of major economic importance. Changes observed by radiography were also studied by conventional parasitological methods. The radiodiagnostic method reported here appears to be a useful complement to diagnostic examinations that have been based merely on dissection so far. It enables evaluation of the pathological lesions in live condition, without causing damage to the fish.     

Activities of biotransformation enzymes in pheasant (Phasianus colchicus) and their modulation by in vivo administration of mebendazole and flubendazole

Basal activities of certain pheasant hepatic and intestinal biotransformation enzymes and modulation of their activities by anthelmintics flubendazole (FLBZ) and mebendazole (MBZ) were investigated in subcellular fractions that were prepared from liver and small intestine of control and FLBZ or MBZ treated birds. Several oxidation, reduction and conjugation enzyme activities were assessed. In the liver, treatment of pheasants by FLBZ or MBZ caused very slight or no changes in monooxygenase activities and conjugation enzymes. More significative changes were detected in small intestine. Metyrapone and daunorubicin reductase activities were increased by both substances in the liver. This is the first evidence that certain benzimidazoles modulate reductases of carbonyl group. With respect to the relatively slight extent of the changes caused by FLBZ or MBZ we can assume that repeated administration of therapeutic doses of both FLBZ and MBZ has probably no serious influence on pheasant biotransformation enzyme system.