Analysis of In vitro Fertilizing Capacity to Evaluate the Freezing Procedures of Boar Semen and to Predict the Subsequent Fertility

A porcine in vitro fertilization (IVF) system and seminal quality parameters of frozen–thawed boar semen were used to assess the effectiveness of two different thawing rates of frozen boar semen, and to address the question of whether differences between fertility of ejaculates could be predicted in a limited field trial. In the first experiment, two thawing procedures were analysed (37°C, 30 s; 50°C, 12 s) and no differences in sperm quality were found. However, when the procedure was 50°C, 12 s the IVF results showed a higher number of sperm per penetrated oocyte and a near 10 points higher rate of pronuclear formation. In the second experiment, the fertility results obtained in the limited field trial show to be efficient enough for application in a commercial use, especially for three of the employed boars (fertility ≥80%). In this limited study, the conventional seminal parameters are not accurate enough to discriminate good and bad boars in relation to fertility. On the contrary, parameters of in vitro penetrability are more precise to predict subsequent fertilities. As conclusion, the IVF fertilization system seems to be a good tool to evaluate the quality of frozen–thawed boar semen previous to its commercial way, to verify the bank semen storage quality and a good way to assay new sperm freezing procedures, as it is the more precise evaluating method in estimating the potential fertilizing ability.     

A specific DNA probe which identifies Babesia bovis in whole blood.

A genomic library of Babesia bovis DNA from the Mexican strain M was constructed in plasmid pUN121 and cloned in Escherichia coli. Several recombinants which hybridized strongly to radioactively labeled B. bovis genomic DNA in an in situ screening were selected and further analyzed for those which specifically hybridized to B. bovis DNA. It was found that pMU-B1 had the highest sensitivity, detecting 25 pg of purified B. bovis DNA, and 300 parasites in 10 microliters of whole infected blood, or 0.00025% parasitemia. pMU-B1 contained a 6.0 kb B. bovis DNA insert which did not cross-hybridize to Babesia bigemina, Trypanosoma evansi, Plasmodium falciparum, Anaplasma marginale, Boophilus microplus and cow DNA. In the Southern blot analysis of genomic DNA, pMU-B1 could differentiate between two B. bovis geographic isolates, Mexican strain M and Thai isolate TS4. Thus, the pMU-B1 probe will be useful in the diagnosis of Babesia infection in cattle and ticks, and in the differentiation of B. bovis strains.     

Use of Echinococcus granulosus worm antigens for immunodiagnosis of E. granulosus infection in dogs.

Echinococcus granulosus worm excretory/secretory antigens (WES) were used in ELISA for diagnosis of E. granulosus infection in dogs and compared with protoscolex somatic antigens (PSM). Sera from 224 dogs were tested. There was no correlation between ELISA absorbance values and E. granulosus worm burdens using either antigen. There was a significant linear relationship between absorbance values of sera tested in the ELISA using WES (W-ELISA) and the ELISA using PSM (P-ELISA). However, there was a small but significant difference between the absorbance values of the sera tested against the two antigens. Western blot analysis of WES using sera from E. granulosus-infected and uninfected dogs revealed antigenic components of relative molecular mass (Mr) larger than 94,000, Mr 94,000-68,000 and Mr 43,000-39,000 in worms, and these were specific for E. granulosus and not identified in PSM; these antigenic differences may be responsible for differences in reactivity in ELISA. The sensitivities of W-ELISA and P-ELISA were 80.8% and 75.6%, respectively. The specificities of W-ELISA and P-ELISA were 93.7% and 97.9%, respectively. The reduced specificity in W-ELISA was mainly attributable to increased background reactivity of sera from Taenia hydatigena-infected dogs. Despite the reduction in specificity, both ELISAs are valuable epidemiological tools to determine the prevalence of antibody to E. granulosus in dog populations and to monitor the success of hydatid control campaigns.     

Surgery of hepatic cysts in a cat.

A 2-year-old female Japanese domestic cat weighing 3.6 kg was presented due to marked abdominal distention and was diagnosed as hepatic cyst. The bile acid concentration of the hepatic cyst was as low as the serum bile acid concentration. The membrane of the cysts was excised from the edge of the compressed hepatic tissue and no other surgical manipulation was performed. This surgical procedure is simple and quick compared to hepatic lobectomy, however, it is necessary to evaluate the fluid of the cyst before performing this procedure.     

A Quantitative Study of Ganglion Cells in the German Shepherd Dog Retina

As in the number of mammals, the most prominent feature of the ganglion-cell layer in the retina of the German shepherd dog is the sharp increase in the density of ganglion cells in the central area. There is an area of maximum density and also a ‘cat-like’ visual streak, located dorsal to the optic disc. The isodensity lines of ganglion-cell distribution is roughly concentric. Their values vary from 5300-13 000 cells/mm² in the central area, with the cells densely packed, to 1000 cells/mm² or less in the periphery, where the cells are sparsely distributed. There were some individual differences amongst the animals studied, although all of them were pure-bred dogs. This suggests that the configuration of the retina in the canine species is not only dependent on the breed itself but also on some other parameters such as phylogenetic heritage, environment, aptitude, lifestyle, or even training.     

Cross protection studies on Bordetella bronchiseptica in mice using an intracerebral challenge model.

Protective activities of heat-inactivated (60 degrees C for 30 min) merthiolate preserved Bordetella bronchiseptica and B. pertussis bacterins were compared in intraperitoneally immunized mice challenged intracerebrally (i.p./i.c.) or intraperitoneally (i.p./i.p.). In the i.p./i.c. assay (Kendrick test), a B. pertussis bacterin protected mice against challenge with B. pertussis 18-323, as well as against phase I cytotoxic and non-cytotoxic strains of B. bronchiseptica. A B. bronchiseptica bacterin, prepared from a phase I cytotoxic strain, gave protection against two phase I B. bronchiseptica strains, irrespective of their cytotoxin-production. A non-cytotoxic phase I strain of B. bronchiseptica elicited protection against the homologous strain only. Neither cytotoxic nor non-cytotoxic B. bronchiseptica strains protected mice challenged with B. pertussis 18-323. Vaccines prepared from phase III strains of B. bronchiseptica were not protective at all against any of the challenge strains. No such differences in the protective activities of the bacterins could be detected by the i.p./i.p. method. They seem to cross-protect equally well. The results indicate that the Kendrick test may be useful in testing potency of different B. bronchiseptica bacterins.     

The diagnostic significance of different blood parameters and liver biopsy in chronic copper poisoning of sheep

In a flock of sheep to which food with excess copper (= Cu) had been fed for some months 39 animals died of chronic copper toxicosis. In 5 sheep of this flock the plasma concentrations of the enzymes glutamate dehydrogenase (GLDH), sorbitol dehydrogenase (SDH), aspartate-amino-transferase (ASAT) and copper were measured several times over a period of 100 days. At day 23, 58 and 97 needle biopsies of the liver were taken. The biopsies were used for histological examination and Cu-analysis. After 100 days the sheep were slaughtered and chronic copper poisoning was diagnosed by raised Cu concentration and typical histological alterations in the liver. The activities of GLDH and SDH in the plasma of the 5 sheep were consistently above normal, ASAT activity and the plasma Cu level were raised only occasionally. The results of Cu determination in biopsy samples were of little predictive value compared with post mortem Cu analysis of the liver.     

Residue management for reducing evaporation in relation to soil type and evaporativity1

Abstract. Usual residue-management options are to remove the residue, use it as mulch with or without undercutting or to incorporate it into the soil. While the role of surface mulch in evaporation has been widely studied, the information on the effect on evaporation of mulch with undercutting or residue incorporated into soil, particularly in relation to soil type and evaporativity (E_o) is lacking. We studied the effect of wheat straw used in various ways on the course of evaporation loss from soil columns with three soils at Ludhiana, India and one soil at Bushland, Texas, USA, under two E_o's Energy-limited evaporation rates under mulch (E_om) followed the soil-specific relation E_om/E_o= a e^(bRes+cEo), where Res is residue rate t/ha and a, b and c are constants; E_o, is expressed in mm/d. In an effort to model the total evaporation (CE) during the energy-limited stage ‘U’ was obtained from appropriate CE versus time curves and (CE-U) was regressed over (t - ti)^0.5 to obtain the slope ‘α’ (Ritchie 1972) for the soil-limited evaporation stage. The observed ‘U’ was independent of mulch rate and E_o but was strongly affected by soil type, Values of ‘α’ decreased with increase in mulch rate and decrease in E_o and coarseness of soil. The otherwise short lived benefit of evaporation reduction with mulch per se, which peaked after a few days was maintained when residue was mixed with soil at the stage when evaporation reduction reached a maximum; this benefit continued for several weeks. Cumulative evaporation values computed from ‘U’ and ‘α’ agreed closely with the observed values under straw mulch for loamy sand and clay loam soils and for ‘undercut’ and ‘residue mixed’ treatments on all soils regardless of E_o, and for all situations under small E_o. However, for sandy loam and silt loam soils under E_o of 10 mm/d, the modified square root of the time function of Jalota et al. (1988) gave a better fit.