Vitrification of Bovine Blastocysts Produced In Vitro Inflicts Selective Damage to the Inner Cell Mass

In contrast to the embryos derived from live animals, the embryos produced in vitro undergo increased damage and reduced survival after cryopreservation, particularly when produced with serum. In medium containing serum, retinoic acid increases cell numbers in the inner cell mass and the trophectoderm without altering their relative proportions in the bovine blastocyst. In this work, in medium without serum, we analyzed the contribution of retinoic acid to the development of blastocyst and survival to vitrification, and found a strong cell reduction in the inner mass when compared to the trophectoderm. Day-6 in vitro -produced morulae were treated for 24 h with retinoic acid (0.7 and 1.4 μ m ) and subsequently cultured without additives for a further 24 h period. Day-8 blastocyst production and cell counts in hatched blastocysts were unaffected by retinoic acid. However, Day-7 expanded, vitrified embryos produced with retinoic acid 1.4 μ m survived at lower rates than controls when cultured after warming. Vitrification greatly reduced cell numbers in the inner mass (p < 0.0001), while cells in the trophectoderm remained unaltered. Differential cell counts analysis in blastocysts should be taken up to replace unspecific determination of total cells to appreciate substantial modifications in their exact terms. The strong reduction we found in the inner cell mass could explain why in vitro survival to cryopreservation is sometimes scarcely informative on the viability of the embryo after transfer to recipients.     

Macrozamia toxicosis in a dog

A case of Macrozamia riedlei seed poisoning is described in a young Dachshund. Vomiting and depression commenced within 6 h of ingestion; other signs that developed included severe hepatopathy, jaundice, abdominal pain that was unresponsive to analgesics, severe gastro-intestinal haemorrhage and thrombocytopenia as well as crystalluria and marrow dyserythropoiesis. The dog was euthanased 6 days after ingestion of the seeds.     

Density-dependent grazing on the extraradical hyphal network of the arbuscular mycorrhizal fungus, Glomus intraradices, by the collembolan, Folsomia candida

The effect of microarthropod density on the relative palatibility of selected microfungi and on the integrity of extraradical arbuscular-mycorrhizal (AM) hyphal networks was examined. A series of microcosm experiments were conducted using two conidial fungi (Alternaria alternata and Trichoderma harzianum), one AM fungus (Glomus intraradices) and the microarthropod Folsomia candida (Collembola). We did not detect a significant effect of athropod density on food preference. Food preferences for conidial over mycorrhizal hyphae were consistent across a wide range of collembolan densities. Collembolan density did, however, have a significant effect on the numbers of extraradical AM hyphae which were severed from the plant root in vitro. Even in the presence of a preferred food source, numbers of severed AM hyphal networks increased with increasing collembolan density. Also, the fecundity of Folsomia candida increased in the presence of preferred food sources. We conclude that microarthropod population densities are influenced by the fungal species available and that the potential of microarthropods to impair the efficacy of AM symbioses is density-dependent. Received: 16 May 1997     

Efficacy of an ivermectin and pyrantel pamoate combination against adult hookworm, Ancylostoma braziliense, in dogs

A chewable tablet incorporating ivermectin and pyrantel was tested in 12 Beagle dogs for efficacy against the adult hookworm, Ancylostoma braziliense. The dogs were administered infective larvae of A braziliense orally. Twenty-one days after infection the dogs were weighed and allocated randomly to receive either an oral treatment with ivermectin and pyrantel in a beef-based chewable tablet or no treatment. The chewable tablet was a commercially available product, which was made to deliver ivermectin at 6 μg/kg and pyrantel at 5.0 mg/kg to each dog. Seven days after treatment the dogs were euthanased, necropsied, and examined for adult hookworms. At necropsy, no adult A braziliense was observed in any of the 6 treated dogs and all 6 dogs that had been left untreated were infected with adult A braziliense (range, 48 to 161). It was concluded that this combination product is 100% efficacious against adult A braziliense.     

Cryptococcosis in seven horses

The clinical, radiographic and post-mortem findings in 6 horses with cryptococcal pneumonia and one horse with an abdominal cryptococcal granuloma are described. In pulmonary cryptococcosis, the lesions were either diffuse and multiple, with bilateral lung involvement, or localised mainly to the dorsocaudal region of one lung. The cases of diffuse multiple cryptococcosis were thought to be associated with haematogenous spread of the fungus after gastrointestinal infection and dissemination from regional lymph nodes. The localised form of the disease was thought to have been associated with inhalation of cryptococci. In all cases of pulmonary cryptococcosis, encapsulated yeast-like organisms were demonstrated in Wright's-stained sediment of tracheal washes. In the horse with the abdominal granuloma, cryptococci were present in a fine needle aspirate sample. Isolates of Cryptococcus neoformans var gattii were recovered from 2 of the 5 horses in which cultures were attempted. In addition to a history of previous illness that may have predisposed to infection, most horses in this report had been in areas in which Eucalyptus camaldulensis, or the closely related E rudis, were growing. In humans, an epidemiological relationship between E camaldulensis and infection with C neoformans var gattii has been suggested. Cases of equine cryptococcosis carry a poor prognosis and treatment was not attempted in any of these cases.     

Real-time polymerase chain reaction monitoring of recombinant DNA entry into soil from decomposing roundup ready leaf biomass

Glyphosate-tolerant, Roundup Ready (RR) soybeans account for about 57% of all genetically modified (GM) crops grown worldwide. The entry of recombinant DNA into soil from GM crops has been identified as an environmental concern due to the possibility of their horizontal transfer to soil microorganisms. RR soybeans contain recombinant gene sequences that can be differentiated from wild-type plant and microbial genes in soil by using a sequence-specific molecular beacon and real-time polymerase chain reaction (PCR). A molecular beacon-based real-time PCR system to quantify a wild-type soybean lectin ( le1) gene was designed to compare amounts of endogenous soybean genes to recombinant DNA in soil. Microcosm studies were carried out to develop methodologies for the detection of recombinant DNA from RR soybeans in soil. RR soybean leaf litterbags were imbedded in the soil under controlled environmental conditions (60% water holding capacity, 10/15 degrees C, and 8/16 h day/night) for 30 days. The soybean biomass decomposition was described using a single-phase exponential equation, and the DNA concentration in planta and in soil was quantified using real-time PCR using sequence-specific molecular beacons for the recombinant cp4 epsps and endogenous soybean lectin ( le1) genes. The biomass of RR soybean leaves was 8.6% less than nontransgenic (NT) soybean leaves after 30 days. The pooled half-disappearance time for cp4 epsps and le1 in RR and of le1 in NT soybean leaves was 1.4 days. All genes from leaves were detected in soil after 30 days. This study provides a methodology for monitoring the entry of RR and NT soybean DNA into soil from decomposing plant residues.     

Assessment of Levels and Potential Health Risk of Heavy Metals in Water and Selected Fish Species from the Benue-Niger River Confluence,Lokoja, Nigeria

The concentrations of cadmium (Cd), lead (Pb), chromium (Cr), copper (Cu), zinc (Zn), and iron (Fe) in water and selected fish species (Oreochromis nilzinc (oticus and Clarias gariepinus) from Benue-Niger River Confluence, Nigeria were investigated. The water and fish sample digests were quantified using an atomic absorption spectrophotometer. The study aimed to assess any possible human health risk associated with water and fish consumption from the selected geographical area. The concentration of the metals in the surface water ranged from 0.007 to 0.039, 0.106 to 0.240, 0.092 to 0.0164, 0.682 to 0.878, 0.058 to 0.309, and 0.136 to 1.645 mg/L for Cd, Cr, Cu, Fe, Pb, and Zn, respectively, while the concentrations of the metals, Cd, Pb, Cr, Cu, Zn, and Fe in these fish species ranged from 0.034 to 0.679, 0.500 to 1.75, 0.152 to 0.351, 0.36 to 1.01, 5.32 to 7.41, and 4.25 to 9.17 mg/kg, respectively. Cd, Cr, and Pb levels in fish were higher than the provisional tolerable weekly intake of 7, 1.5, and 25 μg/kg bw, respectively, according to the World Health Organization. Dry season values of the metals in water samples were higher, and sharptooth catfish had higher amounts of the metals. The hazard quotient (HQ) for Cd indicated a possible potential health concern with fish consumption from this area.