Pregnancy loss in dairy cattle in the Waikato region of New Zealand

AIM: To define the incidence rate of pregnancy loss and risk factors for those losses in pasture-fed dairy cattle in the Waikato region of New Zealand.

METHODS: Cows (n=2,004) from 10 pasture-fed, spring-calving dairy herds in the Waikato were enrolled following confirmation of pregnancy 29–45 days after insemination, for inseminations that occurred within the first 16 days of the seasonal breeding period. Transrectal ultrasonographic examinations for pregnancy were conducted at approximately 6, 8, 10, 14 and 22 weeks gestation, and subsequent calving data were recovered. Pregnancy loss was defined as having occurred when a confirmed pregnancy was not rediagnosed, when gross abortion was detected, or when a cow calved <265 days after the confirmed conception date. Data were analysed using reverse stepwise logistic regression and Cox's proportional hazards analysis.

RESULTS: A total of 128 (6.4%) pregnancy losses were detected. The incidence rate was higher in early compared to late gestation (10.9 vs 2.8 losses/10,000 cow-days between Weeks 6–10 vs Weeks 10–14, respectively; p<0.001). Higher rates of loss were associated with the occurrence of clinical mastitis (Hazards ratio (HR)=1.57; p=0.071), being treated for anoestrus (HR=1.69; p=0.007), and in cows that had calving-to-conception intervals ≤63 days compared with those that had calving-to-conception intervals >92 days (HR=2.49; p=0.06). In addition, the rate of pregnancy loss differed between herds (p=0.05).

CONCLUSIONS: The highest rate of pregnancy loss occurred in early gestation. Clinical mastitis, anoestrus and calving late in the calving season were risk factors for pregnancy loss.

CLINICAL RELEVANCE: Pregnancy diagnosis using ultrasonography can be undertaken from 28 days post-insemination. However, due to the high rate of pregnancy loss at this stage of gestation, herdowners need to be warned of possible losses, and cows should be re-examined to confirm pregnancy before certification of pregnancy status is given.     

Effect of Ascorbic Acid Concentrations,Methods of Cooling and Freezing on Boer Goat Semen Cryopreservation

To improve the Boer goat semen quality during cryopreservation process, three experiments were carried out to investigate the effect of (i) different concentration of ascorbic acid supplementation (ii) rate of cooling with chilled semen characteristics and (iii) method of freezing on post‐thaw Boer goat sperm using Tris‐based extender. Ascorbic acid at 8.5 mg/ml improved the sperm parameters (motility, integrity of membrane and acrosome, morphology and viability), compared to control in cooled samples (p < 0.05). With regard to other concentrations and post‐thawed parameters, ascorbic acid at 2.5–8.5 mg/ml led to higher percentages of sperm motility and integrities of membrane and acrosome when compared to control (p < 0.05). Slow cooling rises to higher percentages of sperm motility, acrosome integrity and viability, in comparison with fast cooling, in terms of cooled and frozen samples (p < 0.05). Programmable freezing method produced the higher percentages of sperm motility, integrities of membrane and acrosome and viability when compared to the freezing method of polystyrene box during goat sperm freezing (p < 0.05). In conclusion, chilled and post‐thawed sperm quality of Boer goat was improved when a Tris‐based extender supplemented with ascorbic acid was used at stages of different cooling rates and freezing methods.     

Radiographic evaluation and treatment of feline dental resorptive lesions.

Many feline resorptive lesions are easily diagnosed by clinical oral examination, whereas others require dental radiographs. Radiographs can reveal the presence of resorption, and often the nature of the resorptive process as well. Removal of affected teeth when they cause discomfort, or of the portion of the tooth causing the discomfort, remains the only treatment that provides long-term resolution. Until we understand the etiology of the inciting causes and of the factors contributing to the progression of resorptive lesions, reliable prevention cannot be offered.     

Comparison of two methods of seminal plasma removal on buffalo (Bubalus bubalis) sperm cryopreservation

Cryopreservation causes damage to spermatozoa, and methods minimizing this damage are therefore needed. Although much discussed, seminal plasma removal has become an alternative to improve sperm quality and viability after freezing and has been applied to different species in attempt to obtain good results. The objective of this study was to evaluate semen quality in buffaloes submitted to two methods for seminal plasma removal (filtration and centrifugation). Semen samples were collected from seven Murrah buffalo bulls (Bubalus bubalis) once a week for 8 weeks. Each ejaculate was divided into three groups: control (presence of seminal plasma), centrifugation and filtration. Sperm kinetics was evaluated with the computer‐assisted sperm analysis (CASA) system. Plasmalemma and acrosomal membrane integrity, mitochondrial membrane potential and reactive oxygen species (ROS) were measured by flow cytometry, and lipid peroxidation was evaluated by the thiobarbituric acid reactive substances (TBARS) assay. Seminal plasma removal did not improve sperm kinetics compared to the control group. Centrifugation increased the number of cells with damaged acrosomal membranes (0.77 ± 0.05) and filtration caused greater plasmalemma and acrosomal membrane damage (22.18 ± 1.07). No difference in the mitochondrial membrane potential was observed between groups. In contrast, ROS production was higher in the centrifugation group compared to the control and filtration groups, although no differences in TBARS formation were detected. In conclusion, seminal plasma removal did not improve the quality of thawed buffalo semen compared to control in terms of sperm kinetics, membrane integrity, mitochondrial membrane potential or lipid peroxidation.     

Characterization of the 1B‐Type ω‐Gliadins from Triticum aestivum Cultivar Butte

ω‐Gliadins were purified from wheat (Triticum aestivum L. ‘Butte’) flour and characterized. Although reversed‐phase HPLC (RP‐HPLC) separated the 1B‐encoded ω‐gliadins into two fractions, 1B1 and 1B2, these fractions had nearly identical amino acid compositions, three similar protein bands in SDS‐PAGE, 10 similar spots in two‐dimensional PAGE, and two similar N‐terminal amino acid sequences. The main components had a range in mass of 48,900–51,500 when estimated by mass spectrometry, significantly less than the mass estimated by SDS‐PAGE. The 1B fractions were digested with thermolysin, the peptides were separated by RP‐HPLC, the peptide mass was determined, and nine peptides from each fraction were sequenced with nearly identical results for the 1B1 and 1B2 digests. A possible consensus sequence of the 1B‐encoded ω‐gliadin internal repeat was QQQXP, where X was F, I, or L in descending order of occurrence. The 1D‐encoded ω‐gliadins were purified by RP‐HPLC as a single fraction that had one band in SDS‐PAGE, two spots in two‐dimensional PAGE, two components with mass of 41,923 and 42,770 detected by mass spectrometry, and two N‐terminal sequences. Circular dichroism (CD) spectra for the 1B and 1D ω‐gliadins were similar and were suggestive of mainly flexible random structure with a significant amount of the left‐handed polyproline II helical conformation in the 1D components.     

The human health implication of the use of antimicrobial agents in animal feeds

Summary

Antimicrobials given in subtherapeutic levels in feed are credited with having contributed to lower cost of meat, milk and eggs. The practice often is associated with the acquisition of resistant enteric flora by the involved animals, and this may in turn contribute to the human reservoir of antimicrobial resistant coliforms and salmonellae. Associated farm workers may transiently acquire resistant flora and on rare occasions develop salmonellosis. Although irrefutable evidence of growth promotant properties of antibiotics in animal feed was provided 30 to 40 years ago, additional studies on mechanisms of the effect are presently needed. It may be possible to identify factors effective in promoting growth without deleterious effects on flora. A national surveillance programme of antimicrobial utilisation (both subtherapeutic and therapeutic) among food producing animals should be established. Molecular epidemiologic research efforts will need to be undertaken to establish whether genetic information of animal origin importantly contributes to the human environmental pool of antimicrobial resistance. In the meantime, it does not appear that the use of drugs as feed additives, while allowing their unrestricted use for therapy in both animals and persons, wouldfavorably influence the problems of antimicrobial resistance of salmonellosis in human populations.     

Sequential extraction and quantitative recovery of gliadins, glutenins, and other proteins from small samples of wheat flour

Methods to sequentially extract and fractionate wheat flour proteins were evaluated to reliably quantify gliadins, glutenins, and albumins/globulins in single flour samples. Compositions of the resulting protein fractions were analyzed by RP-HPLC combined with SDS-PAGE. Unknown proteins were identified by mass spectrometry or N-terminal sequencing. The best separation and recovery of discrete albumin/globulin, gliadin, and glutenin fractions from the same flour sample was achieved by extraction with 0.3 M NaI in 7.5% 1-propanol followed by 2% SDS, 25 mM DTT in 25 mM TRIS, pH 8.0, and precipitation of the solubilized proteins with ammonium acetate/methanol followed by acetone. Average flour composition for the variety Butte86 was 10% albumin/globulin, 40% gliadin, and 48% glutenin. This method should be useful for determining flour composition in diverse samples and evaluating relationships between proteins and end-use functionality.     

Deep hooking,landing success and gear loss using inline and offset circle and J hooks when bait fishing for white sturgeon

The issue of deep hooking is of concern in white sturgeon, Acipenser transmontanus Richardson, fisheries because nearly all anglers use bait with a stationary presentation on the river bottom to catch them, and bait fishing is often associated with higher instances of deep hooking and hooking mortality. Deep hooking rates, landing success and catch rates were investigated for anglers bait fishing for white sturgeon using circle and J hooks with inline and offset alignments fished with both active and passive hook‐setting methods. Anglers hooked 578 white sturgeon and landed 508 fish, ranging in size from 60 to 316 cm total length (mean = 137 cm). Deep hooking rates averaged 0.6% and did not differ between hook types, hook alignments or hook‐setting methods. Landing success (the proportion of hooked sturgeon that were successfully landed) and catch rates were also equivalent between hook types, hook alignments and hook‐setting methods; landing success averaged 88% and catch rates averaged 0.27 fish/hr. Results of this study indicate that deep hooking is rare when angling for white sturgeon using standard bait‐fishing gear regardless of hook‐setting method or whether circle or J hooks were used; regulations restricting hook type in sturgeon bait fisheries are therefore unwarranted.     

Integration of transcriptomic and proteomic data from a single wheat cultivar provides new tools for understanding the roles of individual alpha gliadin proteins in flour quality and celiac disease

The complement of alpha gliadins expressed in a single wheat cultivar was examined by assembling expressed sequence tags (ESTs) from Triticum aestivum cv. Butte 86. Twelve of 19 resulting contigs encoded complete proteins, but only two were identical to proteins reported previously. Eight contained various combinations of epitopes important in celiac disease (CD), while four lacked typical CD epitopes. One alpha gliadin contained an additional cysteine residue that could allow incorporation of the protein into glutenin polymers. In addition, two new types of alpha gliadins ending in GFFGTN and GIMSTN were identified. Based on the number of ESTs, genes encoding proteins that contained the greatest numbers of CD epitopes were expressed at the highest levels. Proteins corresponding to 12 contigs were distinguished in Butte 86 flour by tandem mass spectrometry (MS/MS). Unique peptide tags for individual alpha gliadins, including some that lack CD epitopes and the protein containing the extra cysteine, are reported. The ability to distinguish closely related alpha gliadins by MS/MS makes it possible to explore the roles of individual proteins in flour quality, better define relationships between specific proteins and celiac disease, and devise strategies to reduce the immunogenic potential of wheat flour for patients with CD.