Natural, but not lyophilized, low density lypoproteins were an acceptable alternative to egg yolk for cryopreservation of ram semen

The objective was to evaluate the suitability of using natural or lyophilized low density lipoproteins (LDL), in lieu of whole egg yolk, in extenders for cryopreserving ram semen. Once extragonadal sperm reserves were depleted in 10 fertile Santa Ines cross rams, two ejaculates per ram were collected for cryopreservation. Nine extenders were used: Tris-16% egg yolk extender with 5% glycerol as a control (T1), and substitution of whole egg yolk with 8, 12, 16 or 20% natural LDL (T2-T5, respectively), or with 8, 12, 16, or 20% lyophilized LDL (T6-T9). Semen was diluted to 100 X 10(6) sperm/mL, packaged into 0.25 mL straws, cooled, held at 5 C for 3 h, and then frozen in liquid nitrogen vapor. Immediately after thawing (37 degrees C for 30 s), sperm total and progressive motility, and kinetic parameters were analyzed with computer assisted semen analysis (CASA). Percentage of sperm with plasma membrane functional integrity was assessed by the hypoosmotic swelling test (HOST), sperm membrane physical integrity with propidium iodide (PI), and acrosome integrity with FITC-PSA using an epifluorescent microscope. For all sperm end points, there was no difference between the control and natural LDL treatments (P > 0.05): total motility (T1: 20.9 +/- 11.9 and average of T2-T5: 25.9 +/- 13.6%; mean SD), progressive motility (T1: 6.6 +/- 4.2 and average of T2-T5: 11.7 +/- 7.5%), HOST(+) (T1: 23.7 +/- 6.9 and average of T2-T5: 23.2 +/- 8.7%) and PI(-)/PSA(-) (T1: 13.8 +/- 7.8 and average of T2-T5: 18.1 +/- 7.8%). However, lyophilization was apparently unable to preserve the protective function of LDL; every sperm end point was significantly worse than in the control and natural LDL groups. We concluded that natural LDL was appropriate for cryopreserving ram semen, as it yielded results similar to those obtained with whole egg yolk. (C) 2011 Elsevier Inc. All rights reserved.

Oviductal Fluid Proteins Associated with the Bovine Zona Pellucida and the Effect on In Vitro Sperm-Egg Binding, Fertilization and Embryo Development

Studies have demonstrated that oviductal fluid (ODF) proteins associate with eggs of numerous species including the bovine. In this study, the association of three ODF proteins, the bovine oestrus-associated protein, osteopontin (OPN), lipocalin-type prostaglandin D synthase (L-PGDS), with the bovine zona pellucida (ZP) was demonstrated by immunohistochemistry and western blot. The biological function of ODF derived egg-associated OPN and L-PGDS in sperm binding, fertilization and embryonic development was also explored. In vitro matured bovine oocytes were pre-incubated with ODF collected by cannula from cows in oestrus, or ODF with antibodies to OPN, L-PGDS and bovine serum albumin (BSA). Following incubation, oocytes were inseminated with 1 x 10(5) frozen-thawed spermatozoa, and they were evaluated for sperm binding, fertilization and embryonic development in vitro. Pre-treatment of ODF with antibodies to all of proteins reduced sperm binding to the ZP and fertilization in vitro. Cleavage rates were not significantly different among incubations, but rates of embryo development were significantly decreased. We conclude that antibodies to OPN, L-PGDS and BSA react with oocytes incubated with ODF and inhibit sperm binding, fertilization and embryonic development in vitro, suggesting a potential role of these proteins in these events.

Host preference and host suitability in an egg-pupal fruit fly parasitoid, Fopius arisanus (Sonan) (Hym., Braconidae)

The relative oviposition rate of the parasitoid Fopius arisanus (Sonan) was investigated across three frugivorous tephritid species, Bactrocera tryoni Froggart, Bactrocera jarvisi (Tryon) and Bactrocera cucumis French. Choice and no-choice tests were both used. The suitability of these three species for sustaining larval development and survival to the adult stage was also assessed. Fopius arisanus parasitized all three tephritid species. regardless of the method of exposure, but showed stronger preference for B. tryoni and B. jarvisi over B. cucumis. Superparasitism was extremely rare. Successful development of F. arisanus varied across host species. Bactrocera tryoni yielded significantly more parasitoids than B. jarvisi, but no wasps emerged from B. cucumis puparia. Tests were set up in replicated trials. but results were not homogeneous across trials. We discuss the host relationships of F. arisanus with reference to this variation and in relation to host suitability for larval development.

Effects of dietary level of protein, lysine and methionine and strain of bird on production and egg yolk cholesterol

The effects of dietary level of protein (151, 181 g/kg), lysine (nil, 10g L-lysine hydrochloride/kg) and methionine (nil, 5g DL-methionine/kg) on the production performance and egg yolk cholesterol of two strains of birds were studied for 12 weeks. Birds fed on the high protein diet had higher body weight gain, feed conversion ratio (FCR), rate of lay, egg weight and mass and yolk weight and mass. A high lysine diet decreased feed intake and improved FCR. High dietary level of methionine increased egg yolk cholesterol. There were differences between strains of laying bird in feed intake, rate of lay, egg and yolk weights and egg cholesterol content. It is concluded that strain of bird and dietary level of protein and lysine influenced the production performance of birds. Whilst, egg yolk cholesterol was not reduced by any of the factors studied.

Are wing size, wing shape and asymmetry related to field fitness of Trichogramma egg parasitoids?

The effects of wing shape, wing size, and fluctuating asymmetry in these measures oil the field fitness of T. nr. brassicae and T. pretiosum were investigated. Trichogramma wasps mass-reared on eggs of the factitious host Sitotroga cerealella were released in tomato paddocks and those females ovipositing on Helicoverpo spp. eggs were recaptured. Comparisons of the recaptured group with a sample from the release population were used to assess fitness. Wing data were obtained by positioning landmarks on mounted forewings. Size was then measured as the centroid size computed from landmark distances, while Procrustes analysis followed by principal component analysis was used to assess wing shape. Similar findings were obtained for both Trichogramma species: fitness of wasps was strongly related to wing size and some shape dimensions, but not to the asymmetries of these measures. Wasps which performed well in the field had larger wings and a different wing shape compared to wasps from the mass reared population. Both size and the shape dimensions were linearly associated with fitness although there was also some evidence for non-linear selection on shape. The results suggest that wing shape and wing size are reliable predictors of field fitness for these Trichogramma wasps.

Composition and energy density of eggs from two species of freshwater turtle with twofold ranges in egg size

Lipid, protein, ash, carbohydrate and water content and energy density of eggs were measured from different clutches over a range of egg size in two species of freshwater turtle. Dry egg contents consisted of protein (54-60%), lipid (25-31%) and ash (5-6%) while carbohydrate was found to be negligible (

Effect of type of grain and oil supplement on the performance, blood lipoproteins, egg cholesterol and fatty ccids of laying hens

The effects of type of grain (wheat vs. sorghum) and oil supplement/kg diet [0, 20g olive oil (OL), 20g safflower oil (SO), 10 g OL plus 10 g SO (OLSO)], over a 12-week period on the performance, plasma and lipoproteins lipids [cholesterol (C), triglycerides (TG), phospholipid (P)], and yolk C and fatty acids concentrations of laying hens were studied. Hens fed on the sorghum diet had significantly (P

Lethal and sublethal effects of essential oils from Pittosporum undulatum and Hedychium gardnerianum on the egg parasitoid Trichogramma cordubensis (Hymenoptera: Trichogrammatidae

Dissertação de Mestrado, Biotecnologia em Controlo Biológico, 6 de Junho de 2013, Universidade dos Açores.

Splenomegaly in relation to Schistosoma mansoni egg counts: a population based study

A cross: sectional survey on schistosomiasis was done in Comercinho (Minas Gerais State, Brazil), a town with 1474 inhabitants. Stool (Kato-Katz method) and physical examinations were done on 90% of the population and on 84% of the individuals over 2 years of age, respectively. The ecological and individual (case-control) analysis were used to investigate the relation between splenomegaly and S. mansoni egg counts in different age groups. In the ecological analysis there was a clearly correspondence between higher geometric mean of eggs and higher percentage of splenomegaly in the age groups 5-9 and 10-12 years. In the individual analysis it was found that only in the youngest individuals (5-8 or 5-9 years old) the splenomegaly was related with higher mean egg counts in the feces, having been a tendency to the decrease of excretion of eggs in patients with splenomegaly as the age increased. These results strongly suggest that the ecological data are' better indicator of the severity of schistosomiasis in endemic areas, as the decrease of the egg excretion in patients with splenomegaly may be a confounding variable for the individual analysis.

Dot-ELISA for the detection of IgM and IgG antibodies to Schistosoma mansoni worm and egg antigens, associated with egg excretion by patients

Human schistosomiasis, caused by Schistosoma mansoni, is highly prevalent in Brazil and usually diagnosed by time consuming stool analysis. Serological tests are of limited use in this disease, mainly for epidemiological studies, showing no discrimination between previous contact with the parasite and active infections. In the present study, we standardized and compared a Dot-ELISA for IgM and IgG antibodies against S. mansoni antigens from eggs and worms with a routine IgG and IgM immunofluorescence assay using similar antigens, in the study of sera from 27 patients who had quantified egg stool excretion. The positivity obtained for IgG Dot-ELISA was 96.3% and 88.9% for IgM Dot-ELISA with worm antigen and 92.6% and 90.9% with egg antigen. The IFI presented similar positivities using worm antigen, 92.6% (IgG) and 96.3% (IgM),and lower results with egg antigen, 77.8% (IgG and IgM). The patients studied were divided into two groups according to their egg excretion, with greater positivity of serological tests in higher egg excreters. When comparing the quantitative egg excretion and the serological titers of the patients, we detected a correlation only with IgM Dot-ELISA, with r=0.552 (p=0.0127). These data show that Dot-ELISA can be used for the detection of specific antibodies against S. mansoni in sera from suspected patients or in epidemiological studies and, with further purification of egg antigen and larger samples, IgM Dot-ELISA could be a possible tool for rough estimates of parasite burden in epidemiological studies.

Egg excretion in the initial phase of experimental murine schistosomiasis mansoni: stability and association with worm burden

Stability of faecal egg excretion and correlation with results related to worm burden at the initial phase of schistosomiasis mansoni were observed in two groups of mice infected with different Schistosoma mansoni cercarial burdens, by means of analysis of quantitative parasitological studies and schistosome counts after perfusion. Thus, it may be stated that few quantitative parasitological stool examinations could be sufficient to express the infection intensity at the initial phase, on the same grounds that it was already demonstrated at the chronic phase. Furthermore, it is confirmed that the use of the number of eggs passed in the faeces as a tool to estimate the worm burden at the initial phase of schistosome infection is adequate.

Description of the egg of Anopheles (Anopheles) intermedius (Peryassu, 1908) (Diptera: Culicidae) by scanning electron microscopy

The egg of Anopheles (Anopheles) intermedius (Peryassu, 1908) is described and illustrated with scanning electron micrographs. Literature data on An. (Ano.) maculipes (Theobald, 1903) is provided