Serum antibody profile and reproductive performance during two consecutive pregnancies of cows naturally infected with Neospora caninum.

The objective of this study was to record how the antibody levels change over time during pregnancy in dairy cows naturally infected with the protozoan parasite Neospora caninum, and relate this to the reproductive performance. Eighteen cows with antibodies to N. caninum were serum sampled monthly during their first pregnancy and 13 of them were also followed for a second pregnancy. In all, five pregnancies ended in abortion and two in stillbirth. Antibodies to N. caninum in serum were demonstrated by immune stimulating complex enzyme-linked immunosorbent assay (iscom ELISA). The N. caninum antibody titres remained well above the 1:100 cut-off limit for the test used during 2 years in all cows. In the non-aborting cows, mean values of antibody titres to N. caninum rose 1.5-2.5 dilution steps to reach a plateau 4-5 months before parturition, and thereafter decreased from 2 months before parturition. These changes were statistically significant (p < 0.001). The same pattern was seen in the aborting cows. The consistent pattern of rise in antibody titres observed during both pregnancies in all cows indicated a reactivation rather than a reinfection of the parasite at mid-gestation.     

Cattle grazing drives nitrogen and carbon cycling in a temperate salt marsh

We examined the impact of long-term cattle grazing on soil processes and microbial activity in a temperate salt marsh. Soil conditions, microbial biomass and respiration, mineralization and denitrification rates were measured in upper salt marsh that had been ungrazed or cattle grazed for several decades. Increased microbial biomass and soil respiration were observed in grazed marsh, most likely stimulated by enhanced rates of root turnover and root exudation. We found a significant positive effect of grazing on potential N mineralization rates measured in the laboratory, but this difference did not translate to in situ net mineralization measured monthly from May to September. Rates of denitrification were lowest in the grazed marsh and appeared to be limited by nitrate availability, possibly due to more anoxic conditions and lower rates of nitrification. The major effect of grazing on N cycling therefore appeared to be in limiting losses of N through denitrification, which may lead to enhanced nutrient availability to saltmarsh plants, but a reduced ability of the marsh to act as a buffer for land-derived nutrients to adjacent coastal areas. Additionally, we investigated if grazing influences the rates of turnover of labile and refractory C in saltmarsh soils by adding ¹⁴C-labelled leaf litter or root exudates to soil samples and monitoring the evolution of ¹⁴CO₂. Grazing had little effect on the rates of mineralization of ¹⁴C used as a respiratory substrate, but a larger proportion of ¹⁴C was partitioned into microbial biomass and immobilized in long- and medium-term storage pools in the grazed treatment. Grazing slowed down the turnover of the microbial biomass, which resulted in longer turnover times for both leaf litter and root exudates. Grazing may therefore affect the longevity of C in the soil and alter C storage and utilization pathways in the microbial community.     

Comparison of the hydrolysis of bovine κ-casein by camel and bovine chymosin: a kinetic and specificity study

Bovine chymosin constitutes a traditional ingredient for enzymatic milk coagulation in cheese making, providing a strong clotting capacity and low general proteolytic activity. Recently, these properties were surpassed by camel chymosin, but the mechanistic difference behind their action is not yet clear. We used capillary electrophoresis and reversed-phase liquid chromatography-mass spectrometry to compare the first site of hydrolysis of camel and bovine chymosin on bovine κ-casein (CN) and to determine the kinetic parameters of this reaction (pH 6.5; 32 °C). The enzymes showed identical specificities, cleaving the Phe105-Met106 bond of κ-CN to produce para-κ-CN and caseinomacropeptide. Initial formation rates of both products validated Michaelis-Menten modeling of the kinetic properties of both enzymes. Camel chymosin bound κ-CN with ～30% lower affinity (K(M)) and exhibited a 60% higher turnover rate (k(cat)), resulting in ～15% higher catalytic efficiency (k(cat)/K(M)) as compared to bovine chymosin. A local, less dense negatively charged cluster on the surface of camel chymosin may weaken electrostatic binding to the His-Pro cluster of κ-CN to simultaneously impart reduced substrate affinity and accelerated enzyme-substrate dissociation as compared to bovine chymosin.     

Acid‐extractable potassium in agricultural soils: Source minerals assessed by differential and quantitative X‐ray diffraction

Different extraction methods are used world‐wide in routine soil analysis to estimate long‐term potassium (K) reserves for plants. In Sweden, K extracted with 2 M HCl at 100°C (K_HCl) is frequently used, although with limited understanding of the phases extracted. In the present study, we quantified the effects of this extraction on soil minerals in particle size fractions ranging from clay to sand, and estimated their relative contribution to K_HCl. The study included three Swedish long‐term agricultural field experiments with texture ranging from loamy sand to silty clay, as well as mineral specimens of K feldspar. Total weight loss of particle size fractions was determined, and quantitative and differential X‐ray powder diffraction (QXRD, DXRD), applied on solids before and after extraction, was used to quantify the dissolution of individual mineral phases. QXRD and DXRD included spray‐drying of samples, addition of an internal standard and full pattern fitting, where a combination of mineral‐standard XRD traces was matched with the experimental one. Our results show that K_HCl was primarily associated with clay minerals concentrated in the two finest fractions (2–20 and < 2 μm). Highly expandable and mixed‐layer phyllosilicates were quantitatively the most important minerals dissolved. The K was released from micaceous layers in mixed‐layer phyllosilicates with a vermiculitic character. Whether di‐ or trioctahedral, a shared property of the dissolved phases was that they were rich in Fe. In the loamy sand, the coarser fractions (20–2000 μm), where feldspars were prominent, accounted for 35% of K_HCl. According to DXRD, there was no significant decrease in K feldspars in any of the samples, and K_HCl data for the feldspar specimens suggest that clay minerals contributed at least 70% of K_HCl also in the loamy sand. Our study provides insights about the soil minerals that contribute to the long‐term K delivery capacity of soils and an explanation for the prior observation that K_HCl is a dynamic fraction that can be affected by management.     

Breed influences on in vitro development of abattoir-derived bovine oocytes

Background

There is a discrepancy in the reproductive performance between different cattle breeds. Using abattoir-derived ovaries and data base information we studied the effects of breed on in vitro fertilization and early embryo development.

Methods

The in vitro developmental competence of oocytes from cattle (n = 202) of Swedish Red (SR), Swedish Holstein (SH) and mixed beef breeds was compared, retrospectively tracing donors of abattoir-derived ovaries using a combination of the national animal databases and abattoir information. Age was significantly lower and carcass conformation score was higher in the beef breeds than in the dairy breeds.Cumulus oocyte complexes (n = 1351) were aspirated from abattoir-derived ovaries from animals of known breed (visual inspection confirmed through databases), age (databases), and abattoir information. Oocytes were matured, fertilized (frozen semen from two dairy bulls) and cultured according to conventional protocols. On day 8, blastocysts were graded and the number of nuclei determined.

Results

Cleavage rate was not different between the breeds but was significantly different between bulls. The percentage of blastocysts on day 8 was significantly higher when the oocyte donor’s breed was beef or SR than SH. There was no significant difference in blastocyst grades or stages between the breeds, but the number of nuclei in day 8 blastocysts was significantly lower in SH compared to the beef.

Conclusions

The use of abattoir-derived ovaries from animals whose background is traceable can be a valuable tool for research. Using this approach in the present study, oocyte donor breed was seen to affect early embryo development during in vitro embryo production, which may be a contributing factor to the declining fertility in some dairy breeds seen today.     

Application of an indirect carbon immunoassay (CIA) for the rapid diagnosis of antibody to Toxoplasma gondii in sheep

Carbon immunoassay (CIA), a novel indirect rapid test for Toxoplasma antibody in sheep, was compared with indirect fluorescent antibody assay (IFA). CIA relies on the adherence of carbon particles of India-ink to rabbit immunoglobulin G.l Carbon labelled anti-sheep rabbit IgG was used for the detection of sheep antibody when attached to tachyzoites of Toxoplasma gondii. The result was read in an ordinary light microscope and there was a clearcut difference between negative and positive reactions. Out of a total of 97 sheep sera tested, 15 sera were negative in both tests and 12 were negative in CIA but showed low positive titres in IFA. The remaining 70 sera were positive in both tests but the titres were usually about 3 dilution steps lower when investigated with CIA as compared to IFA.     

Assessing carbon balance trade-offs between bioenergy and carbon sequestration of stumps at varying time scales and harvest intensities

Mitigating climate change and securing a supply of energy may be achieved, in part, by substituting fossil fuels with bioenergy from stumps and roots. However, the use of stumps would result in a reduction of the carbon pool and thus, from a greenhouse gas emission perspective, there is a trade-off between using stumps for bioenergy and retaining them in the carbon pool associated with dead organic matter. The objective of this study was to show whether, from a carbon balance perspective and over the short and long-terms, stumps are more important as a source of energy that replaces fossil fuels, or as a carbon sink.     

Schistosoma mansoni antibodies in tissue specimens demonstrated by a diffusion-in-gel ELISA technique.

A modification of the diffusion-in-gel enzyme-linked immunosorbent assay (DIG-ELISA) for analysis of antibodies in tissue fragments has been elaborated. In this technique, tissue specimens are placed on top of a thin gel layer covering a plastic surface coated with an antigen preparation. The gel thus serves as a medium through which diffusion of antibodies contained in tissue samples may occur. Specimens from skin, lung, liver, kidney, heart and skeletal muscle as well as blood serum and blood sampled onto filter paper were successfully used for analysis of the antibody response to two different antigens (soluble adult worm antigen and soluble egg antigen) during experimental infection of mice with the helminth parasite Schistosoma mansoni. It was concluded that skin and lung tissue reflected the serum antibody response more closely than the other tissues tested, although zone sizes were slightly smaller than those registered for corresponding serum samples. The DIG-ELISA reaction zone size was, as expected, shown to be dependent on the weight (size) of the tissue specimen. The variability of the modified technique was comparable to that found for analysis of serum by the conventional DIG-ELISA technique. It is concluded that the described technique may be used for serological analysis when blood samples cannot be easily obtained, e.g. at examination of animal carcasses or single organs.     

Prevalence and transmission of Neospora caninum within infected Swedish dairy herds

In this long-term study, the changes in within-herd Neospora caninum seroprevalences and the relative importance of horizontal and vertical parasite transmission in Swedish dairy herds were investigated. The within-herd prevalences varied between 5.8% and 65.0% when the herds were first sampled. Comparing the prevalences of year 2000 with those found 2-3 years later, nine out of 14 investigated herds had a similar or lower within-herd prevalence while five herds had an increased portion of seropositive individuals. Three herds were free or almost free from infection at their last sampling. Changes in prevalence were not related to the level of prevalence that each herd had when first sampled. Antibody avidity measurements revealed that the majority of seropositive individuals in all herds were chronically infected. Dam-calf pair investigations and inspection of genealogic trees supplemented with antibody test results showed that vertical transmission was by far the dominant transmission route in all herds. Taken together, these results indicate that the prevalence of N. caninum infection in Swedish dairy cattle can be successfully reduced if actions to eliminate infection in a herd are combined with preventive measures to avoid re-infection.