Phytophthora Species Are Common on Nursery Stock Grown for Restoration and Revegetation Purposes in California.

Phytophthora tentaculata was detected for the first time in North America in 2012 in a nursery on sticky monkeyflower plant (Diplacus aurantiacus) and again in 2014 on outplanted native plants. At that time, this species was listed as a federally actionable and reportable pathogen by the USDA. As a result of these detections, California native plant nurseries were surveyed to determine the prevalence of Phytophthora species on native plant nursery stock. A total of 402 samples were collected from 26 different native plant nurseries in California between 2014 and 2016. Sampling focused on plants with symptoms of root and crown rot. Symptomatic tissue was collected and tested by immunoassay, culture, and molecular techniques (PCR). Identifications were made using sequences from the internal transcribed spacer (ITS) rDNA region, a portion of the trnM-trnP-trnM, or the atp9-nad9 mitochondrial regions. Phytophthora was confirmed from 149 of the 402 samples (37%), and from plants in 22 different host families. P. tentaculata was the most frequently detected species in our survey, followed by P. cactorum and members of the P. cryptogea complex. Other species include P. cambivora, P. cinnamomi, P. citricola, P. hedraiandra, P. megasperma, P. multivora, P. nicotianae, P. niederhauserii, P. parvispora, P. pini, P. plurivora, and P. riparia. A few Phytophthora sequences generated from mitochondrial regions could not be assigned to a species. Although this survey was limited to a relatively small number of California native plant nurseries, Phytophthora species were detected from three quarters of them (77%). In addition to sticky monkeyflower, P. tentaculata was detected from seven other hosts, expanding the number of associated hosts. During this survey, P. parvispora was detected for the first time in North America from symptomatic crowns and roots of the nonnative Mexican orange blossom (Choisya ternata). Pathogenicity of P. parvispora and P. nicotianae was confirmed on this host. These findings document the widespread occurrence of Phytophthora spp. in native plant nurseries and highlight the potential risks associated with outplanting infested nursery-grown stock into residential gardens and wildlands.

Utilization of Soil Solarization for Eliminating Viable Tilletia indica Teliospores from Arizona Wheat Fields.

Studies were conducted in Arizona to determine the efficacy of soil solarization for killing teliospores of the soilborne fungal wheat pathogen Tilletia indica. In a replicated study conducted in each of 3 years, T. indica teliospores and bunted wheat kernels were buried in a Karnal bunt-infested wheat field at depths of 5, 10, and 20 cm. Replicate samples were removed from under a clear plastic solarization cover at 7-day intervals and the number of viable teliospores determined. A rapid decline in teliospore viability occurred at all treatment depths over 38 days, with efficacy comparable with methyl bromide protocols using clear plastic sheeting. Initial viability rates of 43, 71, and 82% germination were reduced to 0.1, 7.7, and 0.2% after 38 days (across all depths) in 2003, 2005, and 2006, respectively. Mean daily maximum soil temperatures at 5 and 20 cm under clear plastic in 2003, 2005, and 2006 were 67, 53 and 60°C and 43, 38, and 43°C, respectively. Under current United States Department of Agriculture disease management strategies, the method may be useful for the rapid deregulation of Karnal bunt-affected fields.

Electrophysiological characteristic of the ventromedial hypothalamic nucleus and lateral hypothalamic area in normally fed and 24 hour food deprived conscious sheep.

We registered telemetrically the spontaneous electrical activity of the hypothalamic ventromedial nucleus (VMH) and the lateral hypothalamic area (LH) in conscious adult sheep-male, with implanted electrodes,-in a state of normal feeding or after 24 hour food deprivation. The frequency of electrical activity, the amplitude of electrical activity and the frequency by Fourier analysis were evaluated. The frequency of electrical activity was found to be approximately the same in the VMH and LH with mean values +/- S.E.M. 11.5 +/- 0.5 Hz in feed sheep. The amplitude of electrical activity in the VMH was higher compared with the LH, with mean absolute values +/- S.E.M. 41.5 +/- 2.7 microV in the VMH and 35.5 +/- 3.2 microV in the LH. The Fourier analysis showed that from the total amount of harmonic frequencies-the first one i.e. the basic frequency represented 48-56%, the rest are higher frequencies. In sheep after 24 hour food deprivation the frequency of electrical activity decreased on 9.4-9.6 Hz in average in both regions under study and the absolute values of the amplitude of electrical activity increased. The mean absolute value +/- S.E.M. was 115 +/- 5.2 microV in the VMH and 101.2 +/- 3.0 microV in the LH. Along with theses changes we found by Fourier analysis that in the total amount of harmonic frequencies the basic frequency decreased by 6% due to the greater amount of higher harmonic frequencies.

Nucleic acid synthesis from blood urea 15N in the sheep.

In experiments on 4 sheep fed on a low protein diet [6.2 g N/day] and given a single i.v. dose of 15N-labelled urea [15 mg 15N/kg body mass], the authors found that, from 0.5 to 6 h, mean 15N incorporation rose progressively in the total rumen fluid nitrogen from 0.23 to 0.44 at. % 15N and in the rumen bacterial nitrogen from 0.11 to 0.51 at. % 15N. Up to 3 h, total nitrogen enrichment was greater (0.5 at. % 15N) than enrichment of bacterial nitrogen (0.28 at. % 15N), but from 3 to 6 h there was little difference between them. The mean 15N values in the nucleic acids isolated from rumen fluid bacteria in samples collected 3 and 6 hours after injecting labelled urea into the blood were 0.15 and 0.19 at. % 15N respectively, in nucleic acids isolated from the liver 0.042 and 0.04 at. % 15N, in the total rumen bacterial nitrogen 0.28 and 0.51 at. % 15N and in the total liver nitrogen 0.11 and 0.11 at. % 15N. It is concluded from the results that blood urea nitrogen is utilized for synthesis of the total nitrogenous substances of the sheep's rumen bacteria and liver far more intensively than for synthesis of the nucleic acids isolated from them. At the same time, it is utilized more intensively for nucleic acid synthesis in the rumen bacteria than in the liver.

Utilization of endogenous urea secreted into the gastrointestinal tract of the sheep.

The possibility of the utilization of endogenous urea secreted into the gastrointestinal tract was studied in sheep to which a single dose of 3 g 15N urea (50.4% enrichment) was administered by slow infusion via a permanent surgical cannula into the caecum. During a 7 days' experiment, 31.15 +/- 1.08% of the dose was excreted in the urine and 19.65 +/- 1.08% in the faeces, while 49.20 +/- 1.49% was retained in the organism. The maximum 15N values in blood from the jugular vein were found 10 h after starting the infusion and a high concentration was maintained for a further 24 h; sixty hours after the infusion, no more 15N was found in the blood. The maximum 15N values in the total nitrogen of the rumen contents were observed 24 h after the infusion and low levels could still be detected on the 6th day of the experiment. Evidence that 15N participates in enterohepatic and ruminohepatic circulation, and that it is also utilized, is based on the finding of the total, but mainly the protein nitrogen concentration in the duodenum. The maximum 15N values in the total nitrogen (0.247 +/- 0.041 At % 15N) and protein nitrogen (0.262 +/- 0.023 At % 15N) of the duodenal contents were found 24 h after the infusion and 15N could still be demonstrated there in the nitrogen fractions on the 5th day of the experiment.

Incorporation of 15N-urea into individual amino acids of rumen bacteria and blood plasma protein in sheep.

Two sheep weighing about 25 kg were fed on rations with similar nitrogen content (about 9 g N/day including 15N from 5 g labelled urea) but with different content of readily digestible carbohydrate. Concentrate ration consisted of maize meal (whole plant) 300 g + barley meal 100 g + mineral mixture 20 g + urea 5 g; fibrous ration--meadow hay 250 g + oat straw 170 g + mineral mixture 20 g + urea 5 g. Retention of 15N was 53.8 and 43.6% of intake in sheep on concentrate or fibrous diet, respectively. Incorporation of 15N into individual amino acids of bacterial protein isolated from the rumen and blood plasma proteins was different, depending on the ratio and amino acid. Enrichment of 15N of the bacterial matter and plasma proteins in both sheep exceeded that of individual amino acids, indicating that urea nitrogen was utilized to a large extent for the synthesis of nitrogen compounds other than amino acids.

Utilization of urea 15N for the synthesis of the rumen microorganisms and blood plasma protein in sheep.

Two sheep were fed a diet containing urea and high level of easy fermentable carbohydrate or crude fibre. Incorporation of urea 15N into the total and bacterial N in rumen and NPN of blood plasma was higher, but lower into plasma protein in the sheep fed the diet with easy fermentable carbohydrate. The analysis of data of the rate of enrichment with 15N in protein and NPN of plasma and rumen liquid suggests that urea nitrogen was fixed transiently into plasma protein and after releasing and recirculation throughout the rumen incorporated permanently into plasma protein.

Electrical activity of the amygdala and the septum of rats on a high fat or a high protein diet.

The authors studied biopotentials in the region of the amygdala and the septum of rats fed on a standard, high protein or high fat diet. During the first 3-6 days after changing from the standard to the high protein or high fat diet, a decrease in the amplitude of electrical activity was found in both the regions in question. After 3 days on the high fat or the high protein diet, an increase was found in the frequency of electrical activity in the amygdala or the septum, according to the type of diet. A study of the amplitude of electrical activity showed that the electrical potential in the septum was always lower than in the amygdala. The frequency spectrum analysis showed a marked change in the superimposed frequency curve only in animals fed on the high fat diet.

Study of electrical activity in the area of the hypothalamic ventromedial nucleus and the lateral hypothalamus of rats fed on a high protein and high fat diet.

The authors studied bioelectric potentials in the area of the hypothalamic ventromedial nucleus and the lateral hypothalamus of rats fed on a standard, a high protein and a high fat diet. On the first 3--6 days after changing from the standard to the high fat and high protein diets, a decrease in the amplitude of electrical activity was recorded in both the areas in question. It was also found that the frequency of electrical activity in the hypothalamic ventromedial nucleus or the lateral hypothalamus rose, after 3 days administration of the high fat or the high protein diet, in correlation to the type of diet, and that, in the frequency spectrum, a change occurred in the proportion of basic frequency in relation to superimposed frequencies distorting it. It was further found that there was a permanent difference between the amplitude of electrical activity in the lateral hypothalamus and the hypothalamic ventromedial nucleus.

Endogenous urea secretion into the sheep gastrointestinal tract.

Urea turnover and the proportion of endogenous urea secreted and excreted in the saliva, the bile, the pancreatic juice and the urine and directly across the wall of the digestive tract was studied in 6 experiments, after a single i.v. dose of labelled 15N, in two adult sheep weighing 49 and 50 kg, with permanent biliary and pancreatic fistulus and with an exteriorized right parotid duct. It was found that, of the total amount of endogenous urea secreted into the animals' digestive tract (0.2694+/-0.0138 mg/min/kg b.w.), 10.27+/-0.94% reached the contents in the saliva, 2.12+/-0.28% in the bile and 0.66+/-0.08% in the pancreatic juice, and that 86.95+/-2.1% was secreted into the gastrointestinal tract, across its wall, from the blood capillaries. Exogenous turnover amounted to 0.3228+/-0.192 mg/min/kg. Of the total amount of 476.6 mg i.v. injected 15N urea, 274.1+/-8.86 mg was excreted in the urine 5.1+/-0.9 mg in the bile, 3.19+/-0.06 mg in the pancreatic juice, 4.96+/-0.76 mg via the right parotid gland and 9.34+/-1.09 mg in the faeces. The results show that the quantitatively most important part of the recirculation of endogenous urea is its passage from the blood across the wall of the gastrointestinal tract into its contents.

Utilization of 15N-urea administered into the sheep small intestine.

The retention and excretion of intrajejunally administered 15N-urea was studied in four experiments on two sheep with a permanently fistulated small intestine. In the first 7 days after the administration of 2 g 15N-urea, 18.26% was excreted in the faeces and 19% in the urine; 62.74% was retained in the organism. Urinary excretion took place mainly on the first day and from the 3rd to the 7th day no 15N was present in the urine. The rate of 15N excretion in the faeces was roughly the same for the first 4 days and then fell; on the 7th day there was no 15N in the faeces. The proportion of 15N-urea retained in the organism and excreted in the urine was 81% showing that urea in the ruminant gastrointestinal tract is largely linked up into metabolic circulation as part of the general exchange of nitrogenous compounds.

Incorporation of intravenously administered urea-15N into sheep plasma proteins and their amide groups.

Two sheep with a low and high nitrogen intake (7.6 and 24 g N/day respectively) were given a single intravenous dose of 15N-labelled urea (15.3 mg 15N/kg b.w.) The findings were as follows. The greater part of non-retained 15N from the administered dose was excreted during the first day after the intravenous administration of 15N-urea. Daily excretion in the faeces amounted to 1.35-2.37% of the 15N in the given dose. With a low N intake, more 15N from the given dose (59.4%) was retained in the N pool than with a high N intake (50.5%). The net passage of 15N into the rumen and 15N incorporation into the amide-N of the plasma proteins was likewise greater. 15N incorporation into the amide-N of the plasma proteins rose steadily for 3 days. The porportion of amidic 15N in the plasma proteins rose steadily for 3 days. The proportion of amidic 15N in the plasma protein total 15N changed on the second and third day after administering 15N-urea from 8% to 16%, with the maximum at the beginning of the second day. The amount of 15N incorporated into the proteins in 1 litre plasma attained up to 3% of the given dose. It is concluded from the results that the synthesis of amino acids and their amide groups is both a quantitatively and a qualitatively important metabolic route for the reutilization of blood urea nitrogen for protein synthesis in ruminants.

Passage of the intravenously administered 15N urea into the digestive tract and its excretion in the sheep.

The experiments performed on two wethers provided with simple rumen cannulas and reentrant cannulas, inserted into the proximal duodenum and ileum, showed a passage of 15N from labelled urea, injected intravenously, from the blood to the digestive tract. The amount of the 15N in the digesta was the highest in duodenum, slightly lower in the rumen and slightly lower in ileum. Approximately 50% of the injected 15N was excreted in urine. The amount of the 15N eliminated with feces was very small; 0.6 to 2.8% of the dose injected per day. About 73--84% of the 15N which passed the duodenum was absorbed in further parts of the digestive tract. It can be concluded that all parts of the digestive tract take part in utilization of the endogenous urea.

Incorporation of N from intravenously administered 15N labelled urea into the bacterial protein in the sheep.

The experiment carried out on two wethers demonstrated that nitrogen of intravenously injected urea, labelled with 15N was incorporated into total and bacterial nitrogen fraction of the digesta flowing through the rumen and duodenum. The amount of 15N in the bacterial fraction flowing throught the rumen and duodenum was relatively low in comparison with the amount of 15N in the total nitrogen (14,8% and 8,1% in the rumen and 6,6% and 7,9% in the duodenum. The ratio of the amount of bacterial-N to total-N in the rumen content (12,7 and 7,5%) was only slightly lower than the ratio of bacterial 15N to total 15N. In the duodenum this ratio was a little higher (8,7 and 10,0%). Blood urea nitrogen was utilized only partly in biosynthesis of bacterial protein. The results showed that only a small amount of blood urea nitrogen retained in the organism was utilized for microbial protein synthesis and the majority in some different way.