Comparison of the fractionation and assay of domestic duck and fowl pituitary gonadotrophins

1. Pituitary glycoproteins from domestic ducks and fowls were fractionated to separate luteinising hormone (LH) and follicle stimulating hormone (FSH) activities using the same chromatographic steps. 2. Fractions were bioassayed for LH using the release of progesterone from fowl granulosa cells and for thyroid stimulating hormone (TSH) by measuring the release of thyroxine in 3-d-old chicks. Follicle stimulating hormone activity was measured either in a cockerel-testes radioreceptor assay or by the release of oestrogen from cultured rat Sertoli cells. 3. Fractions containing predominantly FSH or LH activity were isolated from the fowl glycoproteins. Duck gonadotrophin did not occur in fractions corresponding to those containing fowl FSH. 4. Duck gonadotrophin was found in a fraction corresponding with the most highly purified fowl LH fraction. A duck LH fraction was found with little FSH activity for which there was no corresponding fowl LH fraction. 5. It is concluded that domestic fowl and duck gonadotrophins have different chromatographic properties. Further study is required to determine whether the purified duck gonadotrophin preparation comprises proteins with similar physico-chemical properties but with separate FSH and LH biological activities.     

Fetal growth of beef calves. I. Effect of prepartum dietary crude protein on birth weight, blood metabolites and steroid hormone concentrations

Fifty-nine crossbred heifers (427 kg) bred to one Hereford sire were randomly assigned at 75 d prepartum to two diets. Heifers were individually fed, and diets were isocaloric but contained either a low (LP = 81% NRC, .56 kg/d) or high (HP = 141% NRC, .98 kg/d) level of crude protein. Jugular vein cannulae were inserted into 16 LP and 16 HP heifers at 10 prepartum. Daily preprandial blood samples that were collected until parturition were analyzed for serum estradiol-17 beta (E2), progesterone (P4), glucose (G) and urea nitrogen (UN). Heifers fed LP gained slower than HP-fed heifers before calving (.73 vs 1.02 kg/d; P less than .01); immediate post-calving weights and condition scores were 418 vs 444 kg (P less than .01) and 5.4 vs 6.1 (P less than .01; LP vs HP, respectively). Calf birth weights (35.3 vs 36.1 kg), average calving difficulty score (1.6 vs 1.6) and percent assisted births (35.5 vs 35.7%) did not differ (P greater than .10; LP vs HP, respectively). Prepartum concentrations of UN (6.2 vs 13.5 mg/dl) and G (52.9 vs 58.2 mg/dl) were lower (P less than .05) and P4 (5.94 vs 4.26 ng/ml) was higher (P approximately equal to .07) in LP heifers. Prepartum concentration profiles were related to calving difficulty score (CD, 1 = no assistance to 3 = hard pull) for E2 (CD1 vs CD2 + CD3, P less than .01; CD2 vs CD3, P approximately equal to .01), P4 (CD1 vs CD2 + CD3, P less than .05), G (CD1 vs CD2 + CD3, P less than .05) and UN (CD2 vs CD3, P less than .05). After calving, all dams were maintained together on pasture and supplemented with alfalfa hay and grain mix until adequate range forage was available to maintain weight gains. Dams that were fed LP prepartum gained faster than HP dams during this period (.49 vs .15 kg/d; P less than .01). Prebreeding weights (443 vs 453 kg; LP vs HP) and condition scores (5.1 vs 5.1) did not differ, nor was the postpartum interval affected (44 vs 40 d; LP vs HP). There was no effect of dietary protein on dystocia or postpartum interval, although there were diet-induced differences in body weight and condition of the dams at calving. Results indicate that differences in prepartum profiles of serum steroid hormones and metabolites may be related to dystocia, in addition to relative fetal oversize.     

N-acetyl-beta-D-glucosaminidase index as an early biomarker for chronic kidney disease in cats with hyperthyroidism

Background: Hyperthyroid cats are at risk of developing azotemic chronic kidney disease (CKD) and diagnostic tools currently used to screen for CKD in hyperthyroid cats are either unreliable or impractical.
Hypothesis: Urine N -acetyl-β- d -glucosaminidase index (NAG_i) is a good biomarker for azotemic CKD in hyperthyroid cats.
Animals: Twenty-four newly diagnosed nonazotemic hyperthyroid cats and 10 healthy cats.
Methods: All cats were evaluated for hyperthyroidism at baseline. Hyperthyroid cats were treated with methimazole and reevaluated once euthyroid. At the end of the study, cats were divided into 3 groups: healthy cats, nonazotemic, and azotemic euthyroid cats. Baseline group characteristics were compared to predict azotemic CKD. The influence of treatment on NAG_i was evaluated.
Results: Baseline NAG_i was significantly different among groups ( P = .004). Azotemic cats had a higher median value (13.12 U/g) when compared with healthy cats (1.38 U/g). With NAG_i >2.76 U/g, negative and positive predictive values for development of azotemia were 77.7 and 50%, whereas the combination of a urine specific gravity (USG) ≤1.035 and T₄ >7.80 μg/dL enhanced predictive values to 88.9 and 83.3%, respectively. NAG_i values decreased significantly over time in treated nonazotemic cats.
Conclusions and Clinical Relevance: Baseline NAG_i did not differentiate azotemic from nonazotemic euthyroid cats. NAG_i could be used to assess renal function during medical therapy allowing the clinician to adjust methimazole dosage accordingly. The combination of USG and T₄ could optimize identification of appropriate candidates for permanent treatment of hyperthyroidism.     

Survival of Rhizobium japonicum in soil-sludge environment

The usage of sewage sludge on agricultural lands is an effective and inexpensive practice that provides nutrients for crops. A successful legume crop also depends on the survival of Rhizobium in the soil environment. The number of R. japonicum (USDA 110) in treatment groups containing various soil-to-sludge ratios (control, 13:1, 9:1 and 5:1) during incubation for 1, 21 and 42 days was investigated. The control group contained soil without sludge. Mecklenburg clay and Enon sandy loam soils (both are fine, mixed, thermic, ultic Hapludalfs) were used. All treatments were adjusted to pH 6.7 and brought to 75% of field capacity with 1 ml inoculum (9 × 10⁸ cells ml^? 1) and distilled water. Samples were incubated at 25 C and monitored periodically for the number of surviving R. japonicum (USDA 110) organism by the plant infcction-MPN method. Strains were identified by gel-immunodiffusion. Recovery of rhizobia from both soils was < 1% in all treatment groups after 42 days. However, for control, 13:1, 9:1 and 5:1 groups, the percentage recovery was higher in Enon sandy loams (7.9, 2.3, 2.3 and 2.3%, respectively) at 21 days. Recovery of rhizobia in the 5:1 group from both soils was 7.9% after 1 day, whereas control values were 92%. A decline in rhizobial populations in higher sludge soils may be due to the heavy metals present and available during mineralization of sludge in soils. However, the number of R. japonicum that survived to 21 days was 1.7 × 10₅g^?1 and 1.7 x 10⁶g^?1 for Mecklenburg clay and Enon sandy loam soils with highest sludge, respectively.     

Probing single molecule dynamics

The room temperature dynamics of single sulforhodamine 101 molecules dispersed on a glass surface are investigated on two different time scales with near-field optics. On the 10(-2)- to 10(2)-second time scale, intensity fluctuations in the emission from single molecules are examined with polarization measurements, providing insight into their spectroscopic properties. On the nanosecond time scale, the fluorescence lifetimes of single molecules are measured, and their excited-state energy transfer to the aluminum coating of the near-field probe is characterized. A movie of the time-resolved emission demonstrates the feasibility of fluorescence lifetime imaging with single molecule sensitivity, picosecond temporal resolution, and a spatial resolving power beyond the diffraction limit.     

Effects of site,landscape features,and fire regime on vegetation patterns in presettlement southern Wisconsin

The presettlement tree cover (1831–33) of 3 townships in a southern Wisconsin landscape was analyzed using original survey records. Four forest types were identified: closed forest, open forest, savanna, and prairie. Comparisons of vegetation types and landscape pattern were made between the east and west sides of the Pecatonica River, which bisects the landscape and could have acted as a natural fire barrier. West of the river, presettlement tree species richness and diversity were lower and trees were smaller in diameter and less dense than to the east. The major vegetation types to the west were prairie (42% of landscape) and savanna (40%), both fire-susceptible types. Prairie was more common on gentle slopes than on other landforms. To the east, the landscape was 70% forested (closed plus open forest). Here, prairie was more frequent on steep dry sites. These vegetation differences, including the contrasting landscape placement of prairie, are attributed to distinct site characteristics and to disturbance (fire) regimes, with the west likely having more frequent fires. In terms of the four vegetation types, the east landscape was more homogeneous, being dominated by closed forest (50%). West of the Pecatonica River, the landscape was more heterogeneous because of the high proportion of both prairie and savanna; however, in terms of flammability of vegetation, the west was essentially homogeneous (82% prairie plus savanna).     

Systemic mycosis in a California sea lion (Zalophus californianus) with detection of cystofilobasidiales DNA

A 6-yr-old, intact male California sea lion (Zalophus californianus) with a systemic mycosis died after 5 wk of antifungal drug therapy. Antemortem clinical findings included hind flipper swelling, ring-lesions on skin of the flippers, and dermal nodules that increased in size and number spreading from the hind flippers and ventral abdomen to the foreflippers and muzzle. Lesions were accompanied by severe lymphadenopathy and development of systemic clinical signs despite therapy using itraconazole and later voriconazole. Histopathologic evaluation of biopsies revealed granulomatous dermatitis due to infection by fungus-producing yeast cells in tissue. Isolation attempts, using biopsied skin and tissue samples collected at necropsy, failed to yield growth of a fungus producing yeast cells like those in histologic section. Consensus polymerase chain reaction (PCR) tests of biopsied skin for fungal DNA produced an amplicon having significant sequence identity with a Cystofilobasidiales, a fungus belonging to a subclade that includes several Cryptococcus spp. Histopathologic evaluation of necropsy tissues revealed a systemic mycosis with yeast cells disseminated throughout subcutis, lymph nodes, and viscera. Hepatic necrosis was identified associated with acute liver failure, possibly from the voriconazole administration. This is the first report documenting the clinical presentation, treatment, and pathologic findings of infection associated with Cystofilobasidiales in a marine mammal and serves to expand the understanding of mycoses in pinnipeds.