Oxygen Concentration and Cysteamine Supplementation During In vitro Production of Buffalo (Bubalus bubalis) Embryos Affect mRNA Expression of BCL‐2, BCL‐XL,MCL‐1, BAX and BID

This study examined the effects of O₂ concentration (5% vs 20%) during in vitro maturation (IVM), fertilization (IVF) and culture (IVC) or supplementation of IVM and IVC media with cysteamine (50 and 100 μm , respectively; IVM, IVF and IVC carried out in 20% O₂), on blastocyst rate and relative mRNA abundance of some apoptosis‐related genes measured by real‐time qPCR in immature and in vitro‐matured buffalo oocytes and in embryos at 2‐, 4‐, 8‐ to 16‐cell, morula and blastocyst stages. The blastocyst rate was significantly higher (p < 0.05) while the percentage of TUNEL‐positive cells was significantly lower (p < 0.05) under 5% O₂ than that under 20% O₂. The mRNA expression of anti‐apoptotic genes BCL‐2 and MCL‐1 was significantly higher (p < 0.05) and that of pro‐apoptotic genes BAX and BID was lower (p < 0.05) under 5% O₂ than that under 20% O₂ concentration at many embryonic stages. Following cysteamine supplementation, the blastocyst rate and the relative mRNA abundance of BCL‐XL and MCL‐1 was significantly higher (p < 0.05) and that of BAX but not BID was lower (p < 0.05) at many stages of embryonic development, although it did not affect the percentage of TUNEL positive cells in the blastocysts significantly. The mRNA expression pattern of these genes during embryonic development was different in 5% vs 20% O₂ groups and in cysteamine supplemented vs controls. At the 8‐ to 16‐cell stage, where developmental block occurs in buffalo, the relative mRNA abundance of BCL‐2 and MCL‐1 was highest under 5% O₂ concentration and that of BAX and BID was highest (p < 0.05) under 20% O₂ concentration. These results suggest that one of the mechanisms through which beneficial effects of low O₂ concentration and cysteamine supplementation are mediated during in vitro embryo production is through an increase in the expression of anti‐apoptotic and a decrease in the expression of pro‐apoptotic genes.     

A Bayesian meta-analysis of the effects of administering an intra-vaginal (CIDR) device in combination with other hormones on the reproductive performance of cycling,anoestrous and inseminated cows

AIMS: To evaluate the effectiveness of treatment programmes that included controlled internal drug-releasing (CIDR) devices containing progesterone (P4) in improving synchrony of oe- strus, and conception and pregnancy rates in cycling, anoestrous and inseminated dairy cows, using meta-analysis. To describe the difference in response between cycling and anoestrous cows to CIDR-based synchrony programmes.

METHODS: Scientific papers written in the English language between 1989 and 2002 that investigated the effects of treat- ment programmes including CIDR devices on reproductive per- formance in dairy heifers or lactating dairy cows were identified using a computerised literature search. The criteria for inclusion incorporated evidence that treatment allocation was completely randomised; the population studied was lactating dairy cows; and that data were available on submission, conception and pregnancy rates and their associated measures of variability. Re- productive outcomes from 25 synchrony trials (total n= 11,058 cows) were analysed. Summary measures of the effect of treat- ment on reproductive outcome were assessed using fixed- and random-effects Bayesian meta-analysis models.

RESULTS: Treatment programmes including a CIDR device increased the risk of submission in cycling cows (predicted Bayesian RR=2.86, 95% credible interval 1.46–5.67). Compared with controls, synchrony programmes including CIDR devices in cycling dairy cows had no effect on the risk of conception to first service post-treatment (predicted Bayesian RR=1.00, 95% credible interval=0.80-1.24). Compared with controls, synchrony programmes including CIDR devices had no effect on the risk of pregnancy throughout the mating period (pre- dicted Bayesian RR=1.02, 95% credible interval=0.89-1.17). In anoestrous cows, CIDR treatment had no effect on the risk of conception to first service post-treatment and no effect on the risk of pregnancy throughout the mating period, compared with anoestrous, untreated controls (predicted Bayesian RR=0.91 and 0.97, respectively; 95% credible interval=0.68–1.26 and 0.59-1.60, respectively).

CONCLUSION: The results of this meta-analysis showed that synchrony programmes using CIDR devices combined with other hormones reliably enhanced submission rates in lactat- ing dairy cows. The relatively small number of trials with data suitable for analysis and the heterogeneity of results at the indi- vidual trial level limited our ability to confirm either a beneficial or deleterious effect of treatment on conception or pregnancy rates. Further randomised, controlled trials to evaluate the ef- fectiveness of this form of reproductive therapy in commercial dairy farms are needed.     

The Use of Trypanocides and Antibiotics by Maasai Pastoralists

Information was collected on the use of veterinary drugs by Maasai pastoralists in an area of Kenya where tsetse flies and trypanosomosis occur. Three herds of cattle were followed for between 4 and 5 years and records were kept of every veterinary drug treatment given by the livestock owners. Almost all treatments were either with the trypanocides homidium or diminazene, or with oxytetracycline by intramuscular injection. The rate of trypanocide use varied between 0.66 and 1.56 treatments per animal per year, while oxytetracycline use was between 0.20 and 1.00 treatments per animal per year. Farmers were injecting these drugs in the absence of veterinary supervision, obtaining their supplies mainly from local village shops or informal traders. Underdosing with trypanocides appeared to be uncommon and the indications were that farmers generally gave the drugs at dosage rates above the recommended standard dose. Accurate information on the dose rates of oxytetracycline could not be obtained, but it was noted that in most cases farmers gave a single injection rather than a course of treatment. In a proportion of cases, trypanocides and antibiotics were mixed together before injection. The farmers administered the drugs when disease was recognized and were rarely using trypanocides as prophylactics. Although necessity forces the livestock owners to obtain and use these drugs without veterinary supervision, there are concerns with regard to the possibility of drug misuse and the development of drug resistance.     

Infectious Bursal Disease in New Brunswick

A flock of four week old chickens experienced a disease of sudden onset in which the only symptoms were those of depression shortly before death, and in which the predominant histological lesion was necrosis of lymphocytes in the bursa of Fabricius.

A virus, designated strain Sk-1, was isolated from pooled bursal tissue of affected birds and was serologically identified as a strain of the infectious bursal agent. This virus was chloroform resistant, did not hemagglutinate guinea pig or chicken erythrocytes and did not produce a cytopathic effect in chick embryo tissue cultures. Equivocal results were obtained in filtration studies but the agent was less than 100nm in diameter.

Four week old chicks inoculated with strain Sk-1 developed microscopic lesions in the bursa of Fabricius which were similar to those seen in the original field specimens. Inoculated chick embryos exhibited characteristic macroscopic lesions and necrosis of vascular tissue was a common histological change.

A limited serological survey of local poultry flocks indicated that infection by this agent had occurred in four of the ten flocks examined.

     

Estrous and litter traits in gilts altered by altrenogest, flushing and pubertal status

Influences of estrous synchronization with altrenogest and flushing on reproductive traits in gilts were evaluated in three experiments on two farms. Crossbred gilts were fed altrenogest or altrenogest and an additional 1.55 kg ground sorghum grain for at least 10 d before breeding (flushing), or served as controls. Additional grain for the flushing treatment was provided to gilts from the eighth day of altrenogest treatment until they were detected in estrus. The combination of altrenogest and flushing (on Farm A) increased (P less than .05) litter size when compared with gilts treated only with altrenogest and controls that received neither altrenogest nor flushing. This response was entirely among gilts inseminated at their pubertal estrus. For pubertal gilts fed altrenogest and the flushing treatment, litter traits were similar to other treated or control gilts inseminated at a postpubertal estrus. No treatment effects on litter size were detected for gilts inseminated at a postpubertal estrus. Gilts on Farm B responded differently, with larger litter sizes (P = .08) for those treated with altrenogest and flushing plus altrenogest than for control gilts. Reasons for farm differences might be unidentified genetic or management factors or different seasons of the year when gilts were treated on Farm B (summer) vs Farm A (fall, winter and spring). Our results indicate a marked potential for increasing litter size in gilts mated at their pubertal estrus because their unstimulated ovulation rate (no altrenogest or flushing) did not challenge adequately the biological capacity of their uteri.     

Influence of a controlled internal drug release after fixed-time artificial insemination on pregnancy rates and returns to estrus of nonpregnant cows

We determined whether an ovulatory estrus could be resynchronized in previously synchronized, AI nonpregnant cows without compromising pregnancy from the previous synchronized ovulation or to those inseminated at the resynchronized estrus. Ovulation was synchronized in 937 suckled beef cows at 6 locations using a CO-Synch + progesterone insert (controlled internal drug release; CIDR) protocol [a 100-microg injection of GnRH at the time of progesterone insert, followed in 7 d by a 25-mg injection of PGF(2alpha) at insert removal; at 60 h after PGF(2alpha), cows received a fixed-time AI (TAI) plus a second injection of GnRH]. After initial TAI, the cows were assigned randomly to 1 of 4 treatments: 1) untreated (control; n = 237); 2) progesterone insert at 5 d after TAI and removed 14 d after TAI (CIDR5-14; n = 234); 3) progesterone insert placed at 14 d after TAI and removed 21 d after TAI (CIDR14-21; n = 232); or 4) progesterone insert at 5 d after TAI and removed 14 d after TAI and then a new CIDR inserted at 14 d and removed 21 d after TAI (CIDR5-21; n = 234). After TAI, cows were observed twice daily until 25 d after TAI for estrus and inseminated according to the AM-PM rule. Pregnancy was determined at 30 and 60 d after TAI to determine conception to the first and second AI. Pregnancy rates to TAI were similar for control (55%), CIDR5-14 (53%), CIDR14-21 (48%), and CIDR5-21 (53%). A greater (P < 0.05) proportion of nonpregnant cows was detected in estrus in the CIDR5-21 (76/110, 69%) and CIDR14-21 (77/120, 64%) treatments than in controls (44/106, 42%) and CIDR5-14 (39/109, 36%) cows. Although overall pregnancy rates after second AI service were similar, combined conception rates of treatments without a CIDR from d 14 to 21 [68.7% (57/83); control and CIDR5-14 treatments] were greater (P = 0.03) than those with a CIDR during that same interval [53.5% (82/153); CIDR5-21 and CIDR14-21 treatments]. We conclude that placement of a progesterone insert 5 d after a TAI did not compromise or enhance pregnancy rates to TAI; however, conception rates of nonpregnant cows inseminated after a detected estrus were compromised when resynchronized with a CIDR from d 5 or 14 until 21 d after TAI.     

Bayesian zero-inflated predictive modelling of herd-level Salmonella prevalence for risk-based surveillance

The national control programme for Salmonella in Danish swine herds introduced in 1993 has led to a large decrease in pork-associated human cases of salmonellosis. The pork industry is increasingly focused on the cost-effectiveness of surveillance while maintaining consumer confidence in the pork food supply. Using national control programme data from 2003 and 2004, we developed a zero-inflated binomial model to predict which farms were most at risk of Salmonella. We preferentially sampled these high-risk farms using two sampling schemes based on model predictions resulting from a farm's covariate pattern and its random effect. Zero-inflated binomial modelling allows assessment of similarities and differences between factors that affect herd infection status (introduction), and those that affect the seroprevalence in infected herds (persistence and spread). Both large (producing greater than 5000 pigs per annum), and small herds (producing less than 2000 pigs per annum) were at significantly higher risk for infection and subsequent seroprevalence, when compared with medium sized herds (producing between 2000 and 5000 pigs per annum). When compared with herds being located elsewhere, being located in the south of Jutland significantly decreased the risk of herd infection, but increased the risk of a pig from an infected herd being seropositive. The model suggested that many of the herds where Salmonella was not detected were infected, but at a low prevalence. Using cost and sensitivity, we compared the results of our model based sampling schemes with those under the standard sampling scheme, based on herd size, and the recently introduced risk-based approach. Model-based results were less sensitive but show significant cost savings. Further model refinements, sampling schemes and the methods to evaluate their performance are important areas for future work, and these should continue to occur in direct consultation with Danish authorities.     

Effects of increasing days of exposure to prepartum diets on the concentration of certain blood metabolites in dairy cows

Objective To evaluate the effect of the number of days exposure to a prepartum transition diet on mineral, energy and protein metabolism in dairy cows. Design A prospective cohort study. Procedures The prepartum transition diet consisted of ryegrass pasture, cereal hay, grain, grain by-product, protein meals, BioChlor®, rumen modifiers, minerals and vitamins and contained 13.9% crude protein, 10.0 MJ metabolisable energy/kg and a dietary cation anion difference of −35 meq/kg dry matter. Forty cows were bled bi-weekly from their introduction to the prepartum transition diet until day 35 of lactation. Blood samples were submitted for estimation of a range of metabolites. Cubic smoothed splines were fitted to scatterplots of metabolite concentration as a function of day relative to calving and the area under the curve (AUC) was calculated. Linear regression modelling determined the effect of days exposed, age, body condition score (BSC) and calving day on AUC. Results The prepartum AUC of blood phosphorus and beta-hydroxybutyrate increased and the AUC of blood calcium and cholesterol decreased with increasing days exposed to the diet. The postpartum AUC of beta-hydroxybutyrate and non-esterified fatty acids decreased with increasing days exposed but increased with days exposed × age and days exposed × BCS, respectively. The AUC of other metabolites did not vary significantly with number of days exposed. Conclusions Increasing exposure to the prepartum diet significantly altered the AUC of blood metabolites associated with mineral, energy and protein metabolism in ways consistent with improvements in production and reproduction previously reported and the result suggests links between energy, protein and skeletal metabolism.