A framework for evaluation of on-farm mastitis diagnostics in Australia

Numerous culture-based diagnostics are available on the Australian and international markets for on-farm detection of bacterial pathogens in milk. Use of such diagnostics may provide an opportunity to improve the prudent use of antimicrobials in udder health management. Farms are low-resource settings in terms of diagnostic microbiology capacity. The World Health Organisation has identified criteria for the evaluation of diagnostic tests in low resource settings based on Accuracy, Sensitivity, Specificity, User-friendliness, being Rapid or Robust, Equipment-free and being Deliverable (ASSURED). Here, we review how those criteria can be interpreted in the context of microbiological diagnosis of mastitis pathogens, and how on-farm diagnostics that are currently available in Australia perform relative to ASSURED criteria. This evaluation identifies multiple trade-offs, both with regard to scientific criteria and with regards to convenience criteria. More importantly, the purpose of testing may differ between farms, and test performance should be evaluated relative to its intended use. The ability of on-farm mastitis diagnostics to inform mastitis treatment decision-making in a timely and cost-effective manner depends not just on test characteristics but also on farm-specific pathogen prevalence, and on the farm enterprise's priorities and the farm manager's potential courses of action. With most assay evaluations to date conducted in professional laboratories, there is a surprising dearth of information on how well any of the diagnostic tests perform on-farm and, indeed, of the on-farm decision-making processes that they aim to inform.     

High linoleic acid safflower seed supplementation for gestating ewes: effects on ewe performance, lamb survival, and brown fat stores

Objectives of this study were to determine whether feeding high-linoleic safflower seed to gestating ewes increases cold tolerance and survival in lambs, and whether brown adipose tissue (BAT) stores in lambs are affected by prepartum safflower seed supplementation. In Trial 1, 234 gestating ewes (122 in yr 1 and 112 in yr 2; 75.5 and 81.2 +/- 0.6 kg initial BW for yr 1 and 2, respectively) were allotted randomly to one of two dietary treatments (four pens*treatment(-1)*yr(-1)). Ewes were fed alfalfa-based diets containing (DM basis) either 2.8 (LF) or 5.7% (HF) dietary fat beginning 55 (yr 1) and 42 (yr 2) +/- 1 d prepartum. In Trial 2, 40 Rambouillet cross ewes gestating twins (82.9 +/- 1.7 kg BW) were used in 2 yr (20/yr) and were fed diets containing (DM basis) either 1.9 (LF) or 4.9% (HF) dietary fat beginning 53.4 +/- 1.4 d prepartum. The basal diet was 37.5% each of grass and alfalfa hays and 25% corn silage (DM basis). Cracked safflower seeds (18% CP, 32% fat, 25.6% linoleic acid; DM basis) were used as the supplement in HF, whereas safflower meal and corn were used as the supplement in LF for both trials. At parturition, one lamb from each ewe was selected randomly for slaughter. Perirenal (PR) and pericardial (PC) BAT was excised and weighed, and the carcass was frozen for compositional analysis. In Trial 1, more lambs from HF 0.03; 15.4 vs. 5.8 +/- 2.8%), and dams survived (P = 0.03; 88.4 vs. 78.3 +/- 2.9%), fewer died due to starvation (P = there was a tendency for fewer to die due to pneumonia (P = 0.07; 0.0 vs. 1.7 +/- 0.6%). Ewes fed HF tended to wean more lambs per ewe (P = 0.09; 1.4 vs. 1.2 +/- 0.06) but had similar lamb weight weaned per ewe (P = 0.51; 23.1 +/- 1.22 kg). In Trial 2, prepartum ewe plasma NEFA and glucose concentrations increased with advancing gestation (P < 0.001). Lamb rectal temperature tended (P = 0.08) to be higher in LF lambs and tended (P = 0.06) to increase following parturition. Perirenal BAT weight did not differ among treatments (33.01 +/- 1.66 g; P = 0.28; 0.62 +/- 0.30% BW; P = 0.60). Lambs from LF dams tended (P = 0.08) to have greater PC BAT weight; however, the effect was not significant when expressed as a percentage of BW (0.13 +/- 0.007; P = 0.98). High-linoleic safflower seeds fed during the last 45 d of gestation may be beneficial in improving lamb survivability. Our data do not indicate this response was a result of increased BAT stores. More research is necessary to determine mechanisms that enhance lamb survival when high-linoleic saf-flower seed is fed during gestation.     

Feeding value of field pea as a protein source in forage-based diets fed to beef cattle

Three studies were conducted to evaluate the feasibility of field peas as a protein source in diets for beef cattle. In the first study, 4 cultivars of field pea were incubated in situ to determine rate and extent of CP disappearance. Results indicate that field pea cultivars vary in CP content (22.6, 26.1, 22.6, and 19.4%, DM basis for Profi, Arvika, Carneval, and Trapper, respectively). Soluble protein fraction ranged from 34.9% for Trapper to 54.9% for Profi. Degradable CP fraction was greater (P = 0.01) for Trapper compared with the other cultivars, and no differences (P ≥ 0.25) were observed among Profi, Arvika, and Carneval. Rate of CP degradation differed (P ≤ 0.03) for all cultivars, with Profi being the greatest and Trapper the smallest (10.8, 10.0, 8.1, and 6.3 ± 1.4%/h for Profi, Carneval, Arvika, and Trapper, respectively). Estimated RDP was not different (P = 0.21) for all 4 cultivars. In the second study, 30 crossbred beef steers (301 ± 15 kg) were individually fed and used to evaluate effects of field pea processing (whole, rolled, or ground) on steer performance. Diets contained 40% field pea grain. Growing steers consuming whole field pea had greater ADG (P = 0.08) than those consuming processed field pea (1.69, 1.52, and 1.63 ± 0.05 kg/d, for whole, rolled, and ground, respectively). However, DMI (kg/d and as % of BW) and G:F were not different (P ≥ 0.24). In the third study, 35 individually fed gestating beef cows (694 ± 17 kg) were used to evaluate the use of field pea as a protein supplement for medium quality grass hay (9.3% CP). Treatments consisted of whole field peas at 1) 0 g (CON), 2) 680 g (FP680), 3) 1,360 g (FP1360), and 4) 2,040 g (FP2040), and 5) 1,360 g of 74% barley and 26% canola meal (BCM). Total intake (forage + supplement) of gestating beef cows increased with increasing field pea level (linear, P = 0.01; supplemented vs. nonsupplemented, P = 0.01). In summary, protein quantity and rate of ruminal protein degradation vary across sources of field peas used in this study. Additionally, because of source variability, nutrient analysis and animal requirements should be considered when field pea is incorporated into beef cattle diets. Processing field pea does not improve performance of growing steers. Supplementation of field pea to gestating cows consuming medium-quality grass hay increased total DMI. Overall, our data indicate field pea can be used in a wide variety of beef cattle diets.     

Expression of MHC-I and -II in Uterine Tissue from Early Pregnant Bitches

The aim of the study was to investigate the expression of major histocompatibility complex (MHC)-I and -II in uterine tissues from pregnant and non-pregnant bitches, taken at different time periods after mating. The pregnant bitches were ovariohysterectomized during the pre-implantation (group 1, n = 4), implantation (group 2, n = 7) and placentation stage (group 3, n = 7). Non-pregnant animals in diestrus served as controls (group 4, n = 7). The expression of MHC- I and -II in salpinx, apex, middle horn, corpus uteri and at implantation sites was investigated by immunohistochemistry as well as qualitative and quantitative RT-PCR; MHC-I mRNA was detected in all tissues and with quantitative RT-PCR, and no significant changes were detected until placentation. Immunohistologically, at the apex and corpus site, the average number of MHC-II positive cells increased from the pre-implantation to the post-implantation stage (apex: 1.54 ± 1.21 to 3.82 ± 2.93; corpus: 1.62 ± 1.9 to 5.04 ± 4.95; p < 0.05). The greatest numbers of MHC-II positive cells were observed at placentation sites (6.64 ± 5.9). In parallel, a marked increase in the relative mRNA expression of MHC-II in uterine tissues was assessed from the pre-implantation to the placentation stage (relative to Glycerinaldehyd-3-phosphate-Dehydrogenase (GAPDH): 6.9 ± 9.5, 8.4 ± 5.8, p > 0.05). Immunohistologically, in the salpinx, significantly greater numbers of MHC-II positive cells were found in the tissues of pregnant animals than in the control group (p < 0.05). It is proposed that the increase in MHC-II is pregnancy-related, even though the impact on maintenance of canine pregnancy is still unclear.     

Lectin histochemical pattern on the normal and cystic ovaries of sows

The lectin histochemical pattern (LHP) was characterized and compared in normal and cystic ovaries of sows. Six biotinylated lectins (PNA, SBA, WGA, RCA‐1, DBA and UEA‐1) were used on tissue sections. In the normal ovaries, the reaction to UEA‐1 and SBA was mild to moderate in mesothelial and endothelial cells. RCA‐1 staining was mild to moderate in theca interna of growing follicles, corpora luteum and mesothelium. In addition, this lectin presented strong reaction in endothelial cells, granulosa cells of atretic follicles, zona pellucida of growing follicles and plasma. DBA showed strong intensity in mesothelial and endothelial cells. There was mild to moderate reactivity to WGA in granulosa cells, corpus luteum and theca interna of follicles in development, and moderate in zona pellucida, in granulosa cells of atretic follicles and mesothelium. PNA staining was mild to moderate in oocytes and in the adventitia and media of medullary arteries. Changes in the LHP of the cystic ovaries were noted; however, there were no differences in these findings between the follicular and luteinized cysts. UEA‐1 reactivity in the cystic ovaries was moderately reduced in the mesothelial and endothelial cells, whereas there was mild reduction in the DBA staining in the granulosa cells. Reaction to RCA‐1 and WGA in the cysts also was decreased in theca interna, zona pellucida and granulosa cells of atretic follicles. Furthermore, endothelium and theca interna in the cystic ovaries presented mild reduction of marcation to SBA, whereas there was decreased reactivity to PNA in the oocytes and adventitia and media layers of the medullary arteries. The results of the current study show that cysts modify the LHP in swine ovaries. These changes of glycoconjugates in many ovarian structures could modify diverse process and may be one of the reasons for decreased fertility in sows.     

Vascular Endothelial (VEGF) and Epithelial Growth Factor (EGF) as Well as Platelet‐Activating Factor (PAF) and Receptors are Expressed in the Early Pregnant Canine Uterus

The aim of this study was to investigate the course of expression of platelet‐activating factor (PAF), PAF‐receptor (PAF‐R), epidermal growth factor (EGF), EGF‐R, vascular endothelial growth factor (VEGF), VEGF‐R1 and VEGF‐R2 in uterine tissue during canine pregnancy. For this purpose, 20 bitches were ovariohysterectomized at days 10–12 (n = 10), 18–25 (n = 5) and 28–45 (n = 5) days after mating, respectively. The pre‐implantation group was proven pregnant by embryo flushing of the uterus after the operation, the others by sonography. Five embryo negative, that is, non‐pregnant, bitches in diestrus (day 10–12) served as controls. Tissue samples from the uterus (placentation sites and horn width, respectively) were excised and snap‐frozen in liquid nitrogen after embedding in Tissue Tec^®. Extraction of mRNA for RT‐PCR was performed with Tri‐Reagent. In the embryos, mRNA from all factors except VEGF was detected. In the course of pregnancy, significantly higher expression of PAF and PAFR as well as VEGF and VEGFR2 during the pre‐implantation stage than in all other stages and a strong upregulation of EGF during implantation were characteristic. The course of EGF was in diametrical opposition to the course of the receptor. These results point towards an increased demand for VEGF, EGF and PAF during the earliest stages of canine pregnancy.     

Suture exostosis causing obstruction of the nasolacrimal duct in three horses

CASE HISTORY Three horses were presented with facial swelling and epiphora.

CLINICAL FINDINGS: All horses had facial swellings and radiographic findings consistent with exostosis at the lacrimomaxillary suture, and ipsilateral epiphora. Positive contrast dacryocystorhinography demonstrated that the site of obstruction of the nasolacrimal duct was located where the duct traverses the lacrimomaxillary suture.

DIAGNOSIS: Lacrimomaxillary suture exostosis.

CLINICAL RELEVANCE: Lacrimomaxillary suture exostosis should be considered as a differential diagnosis in cases of epiphora in horses. Dacryocystorhinography, preferably retrograde and normograde, may be used as an aid to diagnosis.     

Penny ice cap cores, baffin island, canada, and the wisconsinan foxe dome connection: two states of hudson bay ice cover

Ice cores from Penny Ice Cap, Baffin Island, Canada, provide continuous Holocene records of oxygen isotopic composition (delta18O, proxy for temperature) and atmospheric impurities. A time scale was established with the use of altered seasonal variations, some volcanic horizons, and the age for the end of the Wisconsin ice age determined from the GRIP and GISP2 ice cores. There is pre-Holocene ice near the bed. The change in delta18O since the last glacial maximum (LGM) is at least 12.5 per mil, compared with an expected value of 7 per mil, suggesting that LGM ice originated at the much higher elevations of the then existing Foxe Dome and Foxe Ridge of the Laurentide Ice Sheet. The LGM delta18O values suggest thick ice frozen to the bed of Hudson Bay.     

IFN‐τ Acts in a Dose‐Dependent Manner on Prostaglandin Production by Buffalo Endometrial Stromal Cells Cultured in vitro

Interferon‐τ (IFN‐τ) has been recognized as the primary embryonic signal responsible for maternal recognition of pregnancy. Uterine endometrium produces both prostaglandin F_2α (PGF_2α) and prostaglandin E₂ (PGE₂). PGF_2α is responsible for the luteolysis; however, PGE₂ favours establishment of pregnancy by its luteoprotective action. In this study, the dose‐response effect of recombinant bovine IFN‐τ (rbIFN‐τ) on prostaglandin (PG) production by buffalo endometrial stromal cells cultured in vitro was studied. Buffalo endometrial stromal cells were isolated by double enzymatic digestion, initially with trypsin III followed by a cocktail of trypsin III, collagenase type II and DNase I and subsequently cultured till confluence. Further, cells were treated with different doses of rbIFN‐τ (0.001, 0.01, 0.1, 1.0 and 10 μg/ml) and keeping a separate set of control. Culture supernatant was collected after 6, 12 and 24 h of treatment. PG levels in the culture supernatant were measured by enzyme immune assay (EIA) and total cellular protein estimated by Bradford method. Results indicated that buffalo endometrial stromal cells following rbIFN‐τ treatment enhanced the secretion of both PGE₂ and PGF_2α, and also its ratio in a strict dose‐dependent manner with a significant increase (p < 0.01) in PGE₂ production at 1 μg/ml dose of rbIFN‐τ and maximal stimulation for both PG was observed at 10 μg/ml. Further, both PG production and its ratio were increased significantly (p < 0.01) in a time‐dependent fashion in all the groups at 6, 12 and 24 h post‐treatment with highest level achieved at 24 h as compared with control. Absolute levels of PGE₂ remained higher than PGF_2α indicating PGE₂ as the major PG produced by endometrial stromal cells. The dose‐dependent response of rbIFN‐τ signifies the importance of optimum concentration of IFN‐τ for the embryonic development especially during the critical period to establish successful pregnancy.