Bacteria wilt of Casuarina equisetifolia in India

Pseudomonas solanacearum (Smith) was consistently isolated from the wilted plants of Casuarina equisetifolia. The isolate gave positive reaction to oxidase, catalase and gas production from nitrate and negative reaction for arginine dihydrolase and acid was produced from various carbohydrates. Wilt produced on tomato, egg plant and capsicum indicated that the pathogen belongs to Race I and Biovar III of P. solanacearum which is a new record from India.     

Beziehungen zwischen Fruchtwechselfolgen einerseits und dem Auftreten der Tomaten-Umfallkrankheit und der Zwiebelwelke sowie den Populationen ihrer Pathogene und des Antagonisten Bacillus subtilis im Boden andererseits

Crop rotation showed a pronounced effect on incidence of tomato damping-off and onion (Fusarium) basal rot as well as on the soil population of the two disease incitants. Out of nine rotation systems, wheat/sorghum and wheat/maize preceding tomato diminished damping-off while clover/maize, lentil/sorghum, wheat/sorghum and wheat/maize preceding tomato diminished damping-off while clover/maize, lentil/sorghum, wheat/sorghum and wheat/maize preceding onions increased basal rot incidence. First crops of the rotation system as well as the second crop were of noticeable effect onRhizoctonia solani andFusarium oxysporum f. sp.cepae as onBacillus subtilis (antagonistic to both fungi) population in soil. Almost all rotations increased population ofF. oxysporum f. sp.cepae andB. subtilis in soil while response ofR. solani was variable. The results show thatBac. subtilis did not correlate with tomato damping-off or onion basal rot. There must be some other factors found in the soil that play a role in effect of crop rotation on disease incidence.     

Rosmarinic acid content and RAPD analysis of in vitro regenerated basil (Ocimum americanum) plants

Various in vitro cultures were established from shoot tips of Ocimum americanum seedlings. Rosmarinic acid content of the in vitro produced plants as well as parent plant were determined by HPLC analysis and subjected to RAPD analysis. MS medium with BA at a concentration of 1 mg/l and 0.25 mg/l IAA supports maximum rosmarinic acid production in plants produced from cultures grown on that medium. RAPD analysis revealed 64 scorable bands from four primers, including six polymorphic bands. The band pattern revealed differences between the parent plant and the in vitro regenerated plants. Certain band changes were found in O. americanum plants regenerated in vitro, suggesting the existence of genetic variation that might affect the biochemical synthesis of plants derived from tissue culture.     

Effect of gamma-irradiated entomopathogenic nematodeSteinernema carpocapsae (Fil.) on the larvae of the potato tuber moth,Phthorimaea operculella (Zell.)

The effect of gamma irradiation on the pathogenicity of the entomopathogenic nematodeSteinernema carposapsae was studied by infecting the larvae of the potato tuber moth,Phthorimaea operculella. Full grown larvae of the host were gamma-irradiated with different doses ranged from 2.5 to 160 Gray (Gy) and subsequently infected with either unirradiated or irradiated infective juveniles (IJs) ofS. carpocapsae using the same doses. For comparison, two groups of unirradiated and irradiated host larvae were left without nematode infection. Another group of larval host was also infected with nematodes either unexposed or exposed to the previous doses.The results revealed that there was a rapid mortality in the irradiated host infected with either unexposed nematodes or nematodes irradiated with different doses similar to the mortality in the nematode treatment alone. On the other hand, the pathogenicity of the nematodes was affected as a result of gamma irradiation with high doses, i. e. irradiation of nematodes delayed their efficiencyvis à vis the unirradiated host. However, lower doses of gamma irradiation exhibited worthily stimulating effect on the pathogenic nematodes, one week post irradiation, as it reduced by half the period of time required for killing all irradiated host larvae with regard to nematode treatment alone. Furthermore, nematode treatment whether applied alone or combined with gamma irradiation appears very promising as compared with single irradiation treatment of the host.With 3 tables     

Photosynthetic responses to phosphorus nutrition in two-year-old maritime pine seedlings

We analyzed processes limiting photosynthesis in two-year-old, container-grown Pinus pinaster Ait. seedlings subjected to phosphorus (P) deficiency. After withholding P for 3 months, seedlings were supplied P at four relative addition rates (0, 0.005, 0.01 and 0.02 day(-1)) in a nutrient recycling system. At Weeks 12 and 22, responses of photosynthesis to CO(2) and irradiance were measured and the following parameters derived: maximal velocity of carboxylation by Rubisco, V(m); apparent quantum efficiency of electron transport, alpha maximal electron transport rate, J(m); stomatal conductance and relative stomatal limitation of photosynthesis. At Week 22, these measurements were combined with concurrent measurements of chlorophyll fluorescence to determine the quantum yield of PSII, and a theoretical partitioning of total light-driven linear electron flow between fractions used to regenerate carboxylated and oxygenated RuBP. After 12 weeks of treatment, needle P concentrations ranged from 0.04 to 0.15 x 10(-2) g g(DW) (-1), and then remained constant until Week 22. Values of J(m), alpha and V(m) increased with increasing needle P concentration (from 30 to 133 &mgr;mol m(-2) s(-1), 0.02 to 0.25 mol mol(-1) and 13 to 78 &mgr;mol CO(2) m(-2) s(-1) at the lowest and highest needle P concentrations, respectively). Under ambient conditions, net assimilation rates in P-deficient seedlings were limited by V(m) under saturating irradiance, and by J(m) under limiting irradiance, but not by triose-P regeneration. There was no detectable change in the partitioning of total light-driven linear electron flow between the fractions used for carboxylation and oxygenation. Predawn photochemical efficiency of PSII was significantly reduced in seedlings with low P concentrations. Although stomatal conductance tended to decrease with decreasing needle P concentration, relative stomatal limitation was not significantly affected. At Week 22, there was an attenuation of the effects of P nutrition on V(m) and an increase in alpha and J(m) that was probably related to cessation of growth and the seasonal decline in natural irradiance.     

Use of serology and real time PCR to control an outbreak of bovine brucellosis at a dairy cattle farm in the Nile Delta region,Egypt

Background

Bovine brucellosis remains one of the most prevalent zoonotic infections affecting dairy cattle in developing countries where the applied control programs often fail. We analyzed the epidemiologic pattern of bovine brucellosis in a dairy cattle herd that showed several cases of abortions after regular vaccination with RB51 (B. abortus vaccine). In 2013 thirty dairy cows, from a Holstein-Friesian dairy herd with a population of 600 cattle, aborted five months post vaccination by a regular RB51 vaccine. Blood samples were drawn from milking cows and growing heifers, as well as heifers and cows pregnant up to 6 months. These samples were collected in June 2013 (n?=?257) and May 2014 (n?=?263) and were tested by real time (rt)-PCR as well as serological tests, in particular Rose Bengal Test (RBT), Enzyme-Linked Immunosorbent Assays (ELISA) and Fluorescence Polarization Assay. Tissue specimens were also collected from an aborted fetus and cultured. Isolates were subjected to bacteriological typing tests at the genus and species levels.

Results

Five months post vaccination with RB51 vaccine, Brucella (B.) DNA was detected in blood samples of cows by rt-PCR. The serological tests also revealed the spread of Brucella field strains within the herd in 2013. Four Brucella isolates were recovered from specimens collected from the aborted fetus. These isolates were typed as follows: one B. abortus RB51 vaccine strain and three isolates of B. abortus field strain. The seropositive cows with positive rt-PCR might indicate an infection by the Brucella field strain; while the positive rt-PCR results from seronegative animals may either be due to circulating RB51 vaccine DNA in vaccinated animals or to circulating field strain in infected animals before seroconversion.

Conclusion

The results herein suggest that PCR can be a good supplementary tool in an outbreak situation, if an assay is available that can differentiate vaccine and field strains with a high analytical sensitivity. We recommend using RBT and ELISA in parallel in outbreak situations, to identify as many infected animals as possible during the initial screenings. This test procedure should be repeated for at least three successive negative tests, with one month interval.

     

Effect of a single injection of gonadotropin-releasing hormone (GnRH) and human chorionic gonadotropin (hCG) on testicular blood flow measured by color doppler ultrasonography in male Shiba goats

Although color Doppler ultrasonography has been used to evaluate testicular blood flow in many species, very little has been done in goat. Eight male Shiba goats were exposed to a single intramuscular injection of either gonadotropin-releasing hormone (GnRH group; 1 µg/kg BW) or human chorionic gonadotropin (hCG group; 25 IU/kg BW). Plasma testosterone (T), estradiol (E2) and inhibin (INH) were measured just before (0 hr) and at different intervals post injection by radioimmunoassay. Testis volume (TV) and Doppler indices, such as resistive index (RI) and pulsatility index (PI) of the supratesticular artery, were measured by B-mode and color Doppler ultrasonography, respectively. The results indicated an increase in testicular blood flow in both groups, as RI and PI decreased significantly (P<0.05), but this increase was significant higher and earlier in hCG group (1 hr) than in the GnRH group (2 hr). A high correlation was found for RI and PI with both T (RI, r= −0.862; PI, r= −0.707) and INH in the GnRH group (RI, r=0.661; PI, r=0.701). However, a significant (P<0.05) correlation was found between E2 and both RI (r= −0.610) and PI (r= −0.763) in hCG group. In addition, TV significantly increased and was highly correlated with RI in both groups (GnRH, r= −0.718; hCG, r= −0.779). In conclusion, hCG and GnRH may improve testicular blood flow and TV in Shiba goats.     

RNAi-mediated knockdown of INHBB increases apoptosis and inhibits steroidogenesis in mouse granulosa cells

Inhibins are members of the TGFβ superfamily and act as suppressors of follicle stimulating hormone (FSH) secretion from pituitary glands via a negative feedback mechanism to regulate folliculogenesis. In this study, the INHBB gene was knocked down by three RNAi-Ready pSIREN-RetroQ-ZsGreen vector- mediated recombinant plasmids to explore the effects of INHBB silencing on granulosa cell (GC) cell cycle, apoptosis and steroid production in vitro. Quantitative real-time polymerase chain reaction, Western blot, flow cytometry and ELISA were performed to evaluate the role of INHBB in the mouse GC cell cycle, apoptosis and steroid production in vitro. The results showed that the relative mRNA and protein expression of INHBB in mouse GCs can be significantly reduced by RNAi with pshRNA-B1, pshRNA-B2 and pshRNA-B3 plasmids, with pshRNA-B3 having the best knockdown efficiency. Downregulation of the expression of INHBB significantly arrests cells in the G1 phase of the cell cycle and increases the apoptosis rate in GCs. This was further confirmed by downregulation of the protein expressions of Cyclin D1, Cyclin E and Bcl2, while the protein expression of Bax was upregulated. In addition, specific downregulation of INHBB markedly decreased the concentration of estradiol and progesterone, which was further validated by the decrease in the mRNA levels of CYP19A1and CYP11A1. These findings suggest that inhibin βB is important in the regulation of apoptosis and cell cycle progression in granulosa cells. Furthermore, the inhibin βB subunit has a role in the regulation of steroid hormone biosynthesis. Evidence is accumulating to support the concept that inhibin βB is physiologically essential for early folliculogenesis in the mouse.     

Effect of Mediterranean saltbush (Atriplex halimus) ensilaging with two developed enzyme cocktails on feed intake,nutrient digestibility and ruminal fermentation in sheep

The aim of this study was to assess the effects of feeding Atriplex halimus (AH) silage treated with two developed enzyme cocktails to sheep on feed intake, nutrient digestibility and ruminal fermentation. The AH silage was treated without or with 2 L of ZAD1^® or ZAD2^®/1000 kg with 5% molasses and ensiled for 30 days. Barley grain (300 g/head/day) was fed as an energy supplement once daily at 10.00 hours and AH silage with or without enzyme treatment was offered ad libitum to animals twice daily at 09.00 and 16.00 hours. Sheep were fed on four experimental forage diets comprised of AH silage and barley (D1), AH silage treated with ZAD1^® and barley (D2), AH silage treated with ZAD2^® and barley (D3) and AH silage treated with a combination of ZAD1^® and ZAD2^® (1:1) and barley (D4). Ensiling AH with enzymes reduced its contents of neutral detergent fiber and acid detergent fiber. The dry matter intake of AH of D2, D3 and D4 decreased (P < 0.001) as compared to D1. However, enzyme‐treated diets had greater total digestible nutrients intake (P < 0.001) as compared to D1. The nutrients digestibility for D2, D3 and D4 were higher than those for D1 (P < 0.001), and were higher for D3 as compared to both D2 and D4. Sheep fed on D3 had highest (P < 0.001) ruminal total volatile fatty acids concentration, ammonia nitrogen concentration and microbial protein yield. It could be concluded that AH silage treated with ZAD1^® or ZAD2^® improved digestibility and rumen fermentation in sheep.     

Study on Nutritional Characteristics of Cottonseed Hull as a Feed of Sheep

In this paper through digestive and metabolic experiments studied the voluntary intake and apparent digestibility of cottonseed hulls (kernel-contained CH,kernel-free CH) and cornstalk on sheep to evaluate the nutrition value and digestion characteristics of cottonseed hulls.12 of adult Xinjiang merino rams were selected and divided into two groups (n=6 each group),and on the basis of 300 g/(head·day) of mixed concentrate,fed with kernel-free CH by 400 (group 1) or 800 (group 2) g/(head·day) respectively,to estimate the digestibility of CH by difference method;As well,another 18 of adult Xinjiang merino rams were selected and divided into three groups (n=6 each group),and on the basis of 300 g/(head·day) of mixed concentrate,cafeteria feeding with kernel-contained CH (group 3),kernel-free CH (group 4) or ground cornstalks (group 5) respectively,to study the effects of three diets on the voluntary intake,digestion and metabolism of sheep.The results showed that the apparent digestibility of the DM,OM,CP,cellulose,hemicellulose and energy of kernel-free CH by sheep were 46.71%,47.37%,-53.40%,66.60%,62.48% and 42.59%,respectively.There were no significant differences in the intake of DM,OM,cellulose,hemicellulose,energy and Ca between kernel-contained CH and kernel-free CH groups (P> 0.05);Intake of CP and P in kernel-contained CH group were extremely significantly higher than that in kernel-free CH group (P< 0.01);Intake of lignin in kernel-contained CH group was extremely significantly lower than that in kernel-free CH group (P< 0.01).There were no significant differences in the apparent digestibility of DM,OM and energy between kernel-contained CH and kernel-free CH groups (P> 0.05).The apparent digestibility of cellulose and hemicellulose in kernel-free CH group were significantly and extremely significantly higher than that in kernel-contained CH group (P< 0.05;P< 0.01),respectively.The apparent digestibility of CP in kernel-contained CH group was extremely significantly higher than that in kernel-free CH group (P< 0.01).The apparent digestibility of DM,OM,CP,cellulose and energy in ground cornstalks group were extremely significantly higher than that in kernel-contained CH and kernel-free CH groups (P< 0.01),however,the apparent digestibility of Ca and P were lower than that in kernel-contained CH and kernel-free CH groups (P> 0.05).Intake and retention of nitrogen in kernel-contained CH group were extremely significantly higher than that in kernel-free CH group (P< 0.01);The was no significant difference in the retention of nitrogen between kernel-free diet group and cornstalk diet group (P> 0.05).It was concluded that CH was a kind of special forage with larger intake,higher digestibility on the fiber substances and energy,but with a serious obstruction on protein digestion;The nutritional value between kernel-free diet and cornstalk diet was almost equal,and with lower nitrogen digestibility on kernel-free diet.