Comparison of Five Radioimmunoassay Systems for PAG Measurement: Ability to Detect Early Pregnancy in Cows

This study was conducted to describe the minimum detection limit, reproducibility, accuracy, specificity and parallelism of different pregnancy-associated glycoprotein radioimmunoassay (PAG-RIA) systems: RIA-497, RIA-706, RIA-780, RIA-809 and RIA-Pool. Their ability to distinguish between non-pregnant and pregnant females at day 30 after artificial insemination (AI) was investigated. The antisera were raised in rabbits against different PAG preparations. All RIA systems proved to be sensitive, repeatable and accurate for measuring PAG concentrations. The dilutions of plasma samples taken at an early stage of pregnancy were found to be parallel to the standard curves. No cross-reaction was observed with different carbohydrates, either with Pregnant Mare Serum Gonadotropin (PMSG) or human Chorionic Gonadotropin (hCG). The concentrations of PAG in pregnant females at day 30 after AI were shown to be higher with the use of antisera R#706, R#780, R#809 and Pool when compared with antiserum R#497. All the RIA systems gave 100% sensitivity and negative predictive values. On the other hand, the use of antisera R#780 and R#809 resulted in lower specificity and positive predictive values. The present study clearly shows that the ability of PAG-RIA systems to diagnose pregnancy specifically at day 30 after AI can be improved by using a combination of antisera raised against different forms of PAG.     

Assessing whether dairy cow welfare is “better” in pasture-based than in confinement-based management systems

ABSTRACT

Consumers perceive pasture-based systems of milk production as natural and therefore better for cow welfare than confinement systems. However both systems are heterogeneous and continually evolving, varying from total confinement to total pasture with many hybrid intermediaries. To compare the welfare of dairy cows in these various systems, we use the three spheres framework, comprising biological functioning, natural behaviour and affective states. Considering biological functioning, pasture-based cows are less at risk of subclinical and clinical mastitis, claw lesions, lameness, metritis, early embryonic mortality, culling and mortality, but at more risk of internal parasitism, malnutrition and delayed onset of oestrous activity postpartum than confined cows. Regarding natural behaviours, pasture-based cows exhibit less agonistic behaviour, better lying behaviour, more normal oestrous behaviours and better synchronicity of behaviours than confined cows. They also have the opportunity to graze, which is one of the main features of the behavioural repertoire of dairy cows, but, they may also experience long periods away from pasture in larger herds, and severe climatic stresses which will become increasingly important as the climate changes. Our current ability to assess the affective state of dairy cows is poor. For example, hunger is an important subjective state that cannot be measured directly. The growing focus on ensuring that animals have lives worth living, means that dairy cows should garner some positive emotions from their lives, and it seems clear that pasture access is essential for this. Clearly measurement of affective state is an important challenge for future dairy cow welfare research. At the extremes of management systems, there can be major differences in animal welfare but in hybrid systems, dairy cows experience elements of both confinement and pasture which may ameliorate the negative effects of each on cow welfare. Ultimately, the optimal system gives cows an element of choice between both environments. Moreover management of the system, whether it is confinement or pastured-based, may be as important as the system of management in ensuring good dairy cow welfare and addressing societal concerns.

Abbreviations: BCS: Body condition score; TMR: Total mixed ration     

Assessing the usefulness of B‐mode and colour Doppler sonography,and measurements of circulating progesterone concentrations for determining ovarian responses in superovulated ewes

The main goal of this study was to assess the usefulness of two imaging modalities, namely the B‐mode and colour Doppler sonography, and serum progesterone (P₄) concentrations for determining the ovarian response in superovulated ewes. Twenty‐four sexually mature Santa Inês ewes underwent the superovulatory treatment consisting of eight injections of porcine FSH (total dose of 200 or 133 or 100 mg; n  =  8 ewes/total dose) given at 12‐hr intervals and initiated 48 hr before CIDR ^® (Pfizer Inc., Auckland, New Zealand) removal. Six days after natural mating, the ovaries of all donor ewes were visualized and examined with transrectal ultrasonography and then with videolaparoscopy to identify and enumerate corpora lutea (CL ) and luteinized unovulated follicles (LUF s). Jugular blood samples were collected just prior to ovarian examinations. The total number of CL (r  =  .78 and 0.83, p  <  .0001) and LUF s (r  =  .74 and 0.90, p  <  .0001) enumerated using the B‐mode and colour Doppler ultrasonographic technique, respectively, were correlated with that ascertained by videolaparoscopy. Circulating concentrations of P₄ were related directly to the number of healthy CL (r  =  .73, p  =  .0002) and inversely to the number of prematurely regressing CL (r  = ?.46, p  =  .03), but the accuracy of predicting the number of short‐lived CL with serum P₄ concentrations was very poor. The present results indicate that ultrasonographic imaging and serum P₄ measurements on the day of embryo recovery are useful indicators of total/normal CL numbers and both ultrasonographic techniques can be used to quantify LUF s in superovulated ewes.     

Comparison of the Ability of Three Radioimmunoassay to Detect Pregnancy-associated Glycoproteins in Bovine Plasma

Pregnancy‐associated glycoproteins (PAGs) constitute a large family of glycoproteins that are synthesized in the superficial layer of the ruminant placenta according to a spatial and temporal expression pattern. When PAGs are released in the maternal blood they can be used for pregnancy diagnosis, pregnancy follow‐up and for the monitoring of the trophoblastic function. Three different radioimmunoassay systems (RIA 1, RIA 2 and RIA 3) using antisera produced against PAG I₆₇ (RIA 1), PAG₅₅₊₆₂ (RIA 2) and PAG₅₅₊₅₉ (RIA 3) were used in this investigation in order to measure the PAG concentration in plasma samples withdrawn from pregnant cows and heifers during different periods following artificial insemination (AI). These systems were able to detect PAG molecules in the maternal blood as early as 21 days after AI in different concentrations (RIA 1: 0.43 ± 0.24 ng/ml, mean ± SD; RIA 2: 0.48 ± 0.24 ng/ml; RIA 3: 0.64 ± 0.37 ng/ml). On days 32 and 42 RIA 2 (4.30 ± 1.32 ng/ml and 5.56 ± 1.95 ng/ml) and RIA 3 (4.17 ± 1.15 ng/ml and 5.60 ± 1.89 ng/ml) presented significantly (p < 0.0001) higher PAG concentrations than those of RIA 1 (2.43 ± 0.81 ng/ml and 4.01 ± 1.48 ng/ml), respectively. After day 21, significant correlations (p < 0.0001; r ≥ 0.929) were determined between the three systems. Additionally the three individual PAG profiles presented in this study showed that PAG molecules secreted in the maternal blood between 21 and 50 days after AI were better recognized by the RIA 2 and RIA 3 systems. This study clearly indicated that the ability of a RIA test to recognize PAG molecules in the maternal blood can be improved by carefully selecting the antiserum.     

Validation of a Field Based Chromatin Dispersion Assay to Assess Sperm DNA Fragmentation in the Bottlenose Dolphin (Tursiops truncatus)

Over the last two decades, there have been significant advances in the use of assisted reproductive technology for genetic and reproductive management of captive dolphin populations, including evaluation of sperm DNA quality. This study validated a customized sperm chromatin dispersion test (SCDt) for the bottlenose dolphin (Tursiops truncatus) as a means of assessing sperm DNA damage both in the field and in the laboratory. After performing the SCDt, two different sperm morphotypes were identified: (i) sperm with fragmented DNA showed large haloes of dispersed DNA fragments emerging from a compact sperm nucleoid core and (ii) sperm containing non‐fragmented DNA displayed small compact haloes surrounded by a dense core of non‐dispersed DNA and protein complex. Estimates of sperm DNA fragmentation by means of SCDt were directly comparable to results obtained following a two‐tailed comet assay and showed a significant degree of correlation (r = 0.961; p < 0.001). This investigation also revealed that the SCDt, with minor modifications to the standard protocol, can be successfully conducted in the field using a LED florescence microscopy obtaining a high correlation (r = 0.993; p = 0.01) between the data obtained in the laboratory and in the field.     

Distribution of thiobacillus ferrooxidans and leptospirillum ferrooxidans: implications for generation of acid mine drainage

Although Thiobacillus ferrooxidans and Leptospirillum ferrooxidans are widely considered to be the microorganisms that control the rate of generation of acid mine drainage, little is known about their natural distribution and abundance. Fluorescence in situ hybridization studies showed that at Iron Mountain, California, T. ferrooxidans occurs in peripheral slime-based communities (at pH over 1.3 and temperature under 30 degreesC) but not in important subsurface acid-forming environments (pH 0.3 to 0.7, temperature 30 degrees to 50 degreesC). Leptospirillum ferrooxidans is abundant in slimes and as a planktonic organism in environments with lower pH. Thiobacillus ferrooxidans affects the precipitation of ferric iron solids but plays a limited role in acid generation, and neither species controls direct catalysis at low pH at this site.     

Deep-Sea coral evidence for rapid change in ventilation of the deep north atlantic 15,400 years Ago

Coupled radiocarbon and thorium-230 dates from benthic coral species reveal that the ventilation rate of the North Atlantic upper deep water varied greatly during the last deglaciation. Radiocarbon ages in several corals of the same age, 15.41 +/- 0.17 thousand years, and nearly the same depth, 1800 meters, in the western North Atlantic Ocean increased by as much as 670 years during the 30- to 160-year life spans of the samples. Cadmium/calcium ratios in one coral imply that the nutrient content of these deep waters also increased. Our data show that the deep ocean changed on decadal-centennial time scales during rapid changes in the surface ocean and the atmosphere.     

Pregnancy-associated Glycoprotein Profile during the First Trimester of Pregnancy in Egyptian Buffalo Cows

Pregnancy-associated glycoprotein (PAG) concentrations were measured in buffalo cows starting from day 28 after breeding. Oestrus was synchronized in 10 buffaloes using two injections of 25 mg prostraglandin (PG)F_{2 α} (Lutalyse^®) at a 11-day interval. Blood sampling was conducted nearly twice weekly. Results indicated that plasma PAG concentrations in non-pregnant buffaloes were low (<0.20 ng/ml) during the whole experimental period (day 28 to 103), while in pregnant animals plasma PAG levels increased from day 28 (4.48 ± 0.92 ng/ml) until day 41 (27.27 ± 6.74 ng/ml), remaining high (20.71 ± 9.20 ng/ml) until day 103. Progesterone levels were significantly (p < 0.0001) higher in pregnant (3.51–4.80 ng/ml) than in non-pregnant buffaloes (0.28–1.52 ng/ml). A significant difference (p < 0.0001) in plasma PAG concentrations between pregnant and non-pregnant animals starting at day 28 after breeding suggests that PAG-radioimmunoassay could be suitable for pregnancy diagnosis in buffaloes during this period. In conclusion, PAG test offers the advantages that it requires a single plasma sample for early pregnancy diagnosis as well as the accuracy of the test for the detection of pregnancy as early as day 28.     

A 0.5-million-year record of millennial-scale climate variability in the north atlantic

Long, continuous, marine sediment records from the subpolar North Atlantic document the glacial modulation of regional climate instability throughout the past 0.5 million years. Whenever ice sheet size surpasses a critical threshold indicated by the benthic oxygen isotope (delta18O) value of 3.5 per mil during each of the past five glaciation cycles, indicators of iceberg discharge and sea-surface temperature display dramatically larger amplitudes of millennial-scale variability than when ice sheets are small. Sea-surface temperature oscillations of 1 degrees to 2 degreesC increase in size to approximately 4 degrees to 6 degreesC, and catastrophic iceberg discharges begin alternating repeatedly with brief quiescent intervals. The glacial growth associated with this amplification threshold represents a relatively small departure from the modern ice sheet configuration and sea level. Instability characterizes nearly all observed climate states, with the exception of a limited range of baseline conditions that includes the current Holocene interglacial.