A hemolytic assay for the measurement of equine complement

A hemolytic assay was developed for the measurement of functional equine complement activity. The assay utilizes antibody sensitized chicken erythrocytes as the target cell and was specific for classical pathway (antibody dependent) complement activity. The assay was found to be reproducible and more sensitive than previous reports using other species of target cells. Total serum complement (CH50) values were determined for five mares and their foals and followed over a period of 3 months.     

A leakiness index for assessing landscape function using remote sensing

The cover, number, size, shape, spatial arrangement and orientation of vegetation patches are attributes that have been used to indicate how well landscapes function to retain, not ‘leak’, vital system resources such as rainwater and soil. We derived and tested a directional leakiness index (DLI) for this resource retention function. We used simulated landscape maps where resource flows over map surfaces were directional and where landscape patch attributes were known. Although DLI was most strongly related to patch cover, it also logically related to patch number, size, shape, arrangement and orientation. If the direction of resource flow is multi-directional, a variant of DLI, the multi-directional leakiness index (MDLI) can be used. The utility of DLI and MDLI was demonstrated by applying these indices to three Australian savanna landscapes differing in their remotely sensed vegetation patch attributes. These leakiness indices clearly positioned these three landscapes along a function-dysfunction continuum, where dysfunctional landscapes are leaky (poorly retain resources). This revised version was published online in July 2006 with corrections to the Cover Date.     

Influence of amount and frequency of protein supplementation to steers consuming low-quality,cool-season forage: intake,nutrient digestibility,and ruminal fermentation

This experiment evaluated the influence of protein supplementation frequency (SF) and amount offered on intake, nutrient digestibility, and ruminal fermentation by rumen-fistulated beef steers consuming low-quality [2.9% crude protein (CP); dry matter (DM) basis], cool-season forage. Seven Angus × Hereford steers (300 ± 27 kg) fitted with ruminal cannulas were randomly assigned to 1 of 7 treatments in an incomplete 7 × 4 Latin square. Treatments, in a 2 × 3 factorial design plus a non-supplemented control (CON), consisted of 2 levels of supplemental soybean meal, 100% (F) or 50% (H) of the estimated rumen-degradable protein requirement, provided daily (D), once every 5 d (5D), or once every 10 d (10D). Experimental periods were 30 d and dry matter intake (DMI) was measured from days 19 to 28. On days 21 (all supplements provided) and 30 (only daily supplements provided; day immediately prior to supplementation for 5D and 10D treatments) ruminal fluid was collected for ruminal pH, ammonia-N (NH₃), volatile fatty acids (VFA), and determination of ruminal fermentation variables. Forage and total DM, organic matter (OM), and nitrogen (N) intake increased with supplementation (P ≤ 0.04). However, a linear effect of SF × amount of supplement interaction was observed for forage and total DM, OM, and N intake (P ≤ 0.04), with each variable decreasing as SF decreased, but the decrease being greater with F vs. H. Apparent total tract DM, OM, and neutral detergent fiber digestibility was not affected by supplementation or amount of supplement provided (P ≥ 0.10). In contrast, N digestibility increased with supplementation and for F vs. H (P < 0.01). Digestibility of DM, OM, and N increased linearly as SF decreased (P ≤ 0.03). When all supplements were provided, ruminal NH₃, total VFA, and molar proportions of all individual VFA increased with supplementation (P ≤ 0.04), whereas acetate:propionate ratio decreased (P < 0.01). When only daily supplements were provided, none of the aforementioned fermentation parameters were affected (P ≥ 0.09). In summary, reducing the amount of supplemental CP provided to ruminants consuming low-quality forages, when supplementation intervals are >5 d, can be a management tool to maintain acceptable levels of DMI, nutrient digestibility, and ruminal fermentation while reducing supplementation cost.     

Influence of amount and frequency of protein supplementation to ruminants consuming low-quality cool-season forages: efficiency of nitrogen utilization in lambs and performance of gestating beef cows

We evaluated the influence of amount and crude protein (CP) supplementation frequency (SF) on nitrogen (N) use by wethers and the performance of late-gestation beef cows. In exp. 1, seven Western whiteface wethers (31.8 ± 1.4 kg) were used in an incomplete 7 × 4 Latin square to evaluate intake and N use. Wethers received one of the seven treatments in a 2 × 3 factorial design containing two levels of supplemental soybean meal offered at a rate of 100% (F) or 50% (H; 50% of F) of the estimated CP requirement daily, once every 5, or once every 10 d, plus a non-supplemented control (CON). Low-quality cool-season forage (4.9 % CP; dry matter [DM] basis) was provided daily for ad libitum intake. Experimental periods lasted 30 d. In exp. 2, 84 Angus × Hereford cows (560 ± 35 kg) were stratified by age, body condition score (BCS), and expected calving date and allocated to 1 of the 21 feedlot pens (three pens per treatment). Pens were randomly assigned to receive the same treatments as in exp. 1 and cows had free access to low-quality cool-season forage (2.9% CP; DM basis). Cow body weight (BW) and BCS were measured every 14 d until calving and within 24 h after calving. In exp. 1, supplementation did not alter total DM and organic matter (OM) intake (P ≥ 0.26), but both parameters linearly decreased as SF decreased (P = 0.02). Supplementation increased DM, OM, and neutral detergent fiber (NDF) digestibility (P ≤ 0.02). Additionally, F feeding linearly increased DM, OM, and NDF digestibility as SF decreased (P ≤ 0.04). Digestibility of N, N balance, and digested N retained were greater with supplementation (P < 0.01), and N digestibility linearly increased as SF decreased (P = 0.01). Mean plasma urea-N concentration was not only greater (P < 0.01) for supplemented vs. CON wethers but also greater (P = 0.03) for F vs. H. In exp. 2, pre-calving BCS change was greater (P = 0.03) for supplemented cows. A linear effect of SF × supplementation rate for pre-calving BCS change was noted (P = 0.05), as F-supplemented cows lost more BCS compared with H as SF decreased. When considering supplementation intervals greater than 5 d, reducing the quantity of supplement provided, compared with daily supplementation, may be a feasible management strategy to maintain acceptable nutrient use and animal performance while reducing supplement and labor costs.     

Immunohistochemical expression of HER - 2 and Ki - 67 in granulosa cell tumor in bitches

Granulosa cell tumour, an ovarian neoplasm of stromal origin, is an important tumour related to oestrogenic dominance syndrome and cystic endometrial hyperplasia–pyometra complex. In order to analyse ovarian tumour´s malignant potential, immunohistochemical markers can be used, such as anti-HER2 and anti-Ki-67. The aim of this study was to evaluate the expression of immunohistochemical markers HER-2 and Ki-67 in granulosa cell tumour from bitches´ ovaries. In HER-2 immunomarker analysis using the HercepTest^® method, most tumours were classified as 2+ (moderate labelling). Concerning Ki-67 immunomarker, only one case was described as having a high proliferative index. An association was found between immunostained cell percentage by anti-HER-2 antibodies and high pleomorphism, represented by the pattern of follicular/trabecular tumour arrangement. There was no correlation between anti-Ki-67 and anti-HER-2 antibody immunostaining intensities, probably due to only one case with a high Ki-67 index. With an effective protocol for HER-2 and Ki-67 immunohistochemical identification in granulosa cell tumours in bitches, it was possible to characterize this neoplasm proliferation profile.     

Validation of a multiplex PCR assay to detect Babesia spp. and Anaplasma marginale in cattle in Uruguay in the absence of a gold standard test

Detection of bovine Babesia spp. and Anaplasma marginale is based on the reading of Giemsa-stained blood or organ smears, which can have low sensitivity. Our aim was to improve the detection of bovine Babesia spp. and A. marginale by validating a multiplex PCR (mPCR). We used 466 samples of blood and/or organs of animals with signs and presumptive autopsy findings of babesiosis or anaplasmosis. The primers in our mPCR amplified the rap-1a gene region of Babesia bovis and B. bigemina, and the msp-5 region of A. marginale. We used a Bayesian model with a non-informative priori distribution for the prevalence estimate and informative priori distribution for estimation of sensitivity and specificity. The sensitivity and specificity for smear detection of Babesia spp. were 68.6% and 99.1%, and for A. marginale 85.6% and 98.8%, respectively. Sensitivity and specificity for mPCR detection for Babesia spp. were 94.2% and 97.1%, and for A. marginale 95.2% and 92.7%, respectively. Our mPCR had good accuracy in detecting Babesia spp. and A. marginale, and would be a reliable test for veterinarians to choose the correct treatment for each agent.