小麦钙调素新亚型TaCaM5的克隆及表达分析

利用RT-PCR技术,从条锈菌诱导的小麦叶片中分离出一个编码CaM基因的cDNA序列, 经氨基酸序列分析确定其为一个新的小麦CaM亚型,暂被命名为TaCaM5。TaCaM5包含一个完整450 bp的开放阅读框,编码149个氨基酸;编码的蛋白不含跨膜区、无信号肽、定位在胞内,具有4个EF-hand保守结构域。在目前已知的CaM基因中,TaCaM5与玉米CaM基因的亲缘关系最近,相似性高达97%。该基因在根、茎、叶等组织中均有不同程度的表达;并且受条锈菌诱导表达,在非亲和组合与亲和组合中,分别在接种后6 h和24 h表达量最高。外源植物激素脱落酸、茉莉酸甲酯和乙烯诱导TaCaM5上调表达,水杨酸诱导其下调表达。TaCaM5在机械伤害、干旱和低温条件下表达量上升,在高盐环境下表达量降低。表明TaCaM5可能通过茉莉酸和乙烯等信号途径参与小麦对条锈菌的防御反应,同时参与机械伤害、低温和干旱环境下的Ca²⁺-CaM信号转导途径。     

Identification of the SP22 Sperm Protein in Santa Inês and Dorper Rams

The sperm membrane protein referred to as SP22 has been identified in different species and, at least in rats, is highly correlated with fertility. The goals of this study were to identify and to quantify the SP22 protein on spermatozoa from adult rams (Dorper and Santa Inês breeds), and to correlate its levels to morphological and kinematics parameters. SP22 on ram sperm was effectively quantified by both enzyme‐linked immunosorbent assay (ELISA) and fluorescein isothiocyanate immunostaining analysis and the two methods were significantly correlated (R² = 0.70). Clustering analysis of motility parameters obtained by computer‐assisted semen analysis system was used to establish that three distinct kinematic subpopulations with different vigour and progressiveness coexistent within ejaculate. While there were significant differences in the distribution of the three subpopulations in the rams, there was no significant correlation between the proportion of each subpopulation in the rams and the SP22 levels. Quantification of SP22 immunostaining intensity was not correlated with any of the sperm parameters. However, SP22 levels obtained by ELISA were negatively correlated with morphological abnormalities and positively correlated with membrane integrity (three variable R² = 0.47). Future breeding studies are now needed to validate that this protein is a biomarker of fertility in this species.     

High Periconceptional Protein Intake Modifies Uterine and Embryonic Relationships Increasing Early Pregnancy Losses and Embryo Growth Retardation in Sheep

The effects of supplemented protein level (PL) during the periconceptional period and their interaction with body condition were evaluated in sheep. Multiparous Rambouillet ewes (n = 12) received two PL of rumen undegradable protein (UIP) during a 30‐day pre‐mating and 15‐day post‐mating period: low [LPL, 24% crude protein (CP), 14 g UIP and 36 g/CP animal/day] and high [HPL, 44% CP, 30 g UIP and 50 g/CP animal/day]. While ovulation rate (OR) did not differ between treatments (1.6 ± 0.5, mean ± SEM), a lower fertility rate, a decreased embryo number and a reduced uterine pH (UpH) was observed in the HPL group (p < 0.05), irrespective of BC. Luteal tissue weight, volume and progesterone secretion did not differ among treatments. Sheep with lower UpH also had lower conceptus weight (Cwt; p < 0.05, r = 0.65) and conceptuses with lower mass tended to secrete less INF‐τ and IGF‐1, and the correspondent endometrial explants had a higher basal PGF_2α release. Current study indicates that high protein diets during the periconceptional period in sheep modify uterine and embryonic relationships, increasing early pregnancy losses and inducing embryo growth retardation. Surviving embryos were affected by weight reductions, which could compromise later foetal growth and birth weight. Results evidence the key role of a balanced diet in reproductive success and indicate that the quality and nutrient composition of the maternal diet are essential for an adequate establishment of pregnancy, having paramount effects on the interplay of the embryo and the uterus.     

Dom34:Hbs1 promotes subunit dissociation and peptidyl-tRNA drop-off to initiate no-go decay

No-go decay (NGD) is one of several messenger RNA (mRNA) surveillance systems dedicated to the removal of defective mRNAs from the available pool. Two interacting factors, Dom34 and Hbs1, are genetically implicated in NGD in yeast. Using a reconstituted yeast translation system, we show that Dom34:Hbs1 interacts with the ribosome to promote subunit dissociation and peptidyl-tRNA drop-off. Our data further indicate that these recycling activities are shared by the homologous translation termination factor complex eRF1:eRF3, suggesting a common ancestral function. Because Dom34:Hbs1 activity exhibits no dependence on either peptide length or A-site codon identity, we propose that this quality-control system functions broadly to recycle ribosomes throughout the translation cycle whenever stalls occur.     

Lack of association between Flavobacterium columnare genomovar and virulence in hybrid tilapia Oreochromis niloticus (L.) × Oreochromis aureus (Steindachner)

Columnaris disease can be problematic in tilapia (Oreochromis spp.) production. An understanding of the pathogenesis and virulence of Flavobacterium columnare is needed to develop prevention strategies. The objective of this study was to determine the virulence of genetically defined isolates of F. columnare in sex‐reversed hybrid tilapia, Oreochromis niloticus (L.) × O. aureus (Steindachner). A series of immersion challenge trials were performed using isolates of the five established genomovars of F. columnare: I, II, II‐B, III and I/II. The mean per cent mortality of fish challenged with genomovar I, II and III isolates ranged from 0 to 100, 3.3–78 and 3.3–75%, respectively. The mean per cent mortality of fish challenged with genomovar II‐B ranged from 35 to 96.7%, and the only genomovar I/II isolate tested caused no mortality. Contrary to previous work in other fish species, there did not appear to be an association between F. columnare genomovar and virulence in tilapia. The challenge model used resulted in acute mortality. An alternative challenge model was tested by cohabitating healthy fish with dead fish infected with F. columnare. This method resulted in rapid appearance of clinical signs and mortality, suggesting the potential for F. columnare to increase in virulence upon growth on/in a fish host.     

Effects of thyroxine administration on the growth and survival of pike silverside (Chirostoma estor) juveniles

The pike silverside has high aquaculture potential despite its slow growth. In this study, thyroxine (T₄) concentrations (0.13, 1.3 and 13 nM), a control with no hormonal supplement and a negative control containing 4.5 mM methimazole (MMI) were tested to evaluate the growth of this species. Juveniles (0.2 g) were exposed by immersion to these treatments for 8 h every second day for 120 days, and growth evaluations were performed monthly over the entire trial period. In addition, tissue samples from each treatment group were assayed for triiodothyronine (T₃) and deiodinase type 2 (D2) activity . The survival rates in all T₄ groups were high, and a significant increase in growth was observed (average of 58%). The MMI treatment caused an increase in mortality and a reduction in the final body weight compared with the control. T₄ administration did not affect the tissue levels of T₃, and it decreased muscular D2 activity only after 30 days of exposure. These results demonstrated that low concentrations of T₄ in the culture environment could improve the growth of this species without affecting tissue hormone levels. The technique may have useful applications for early‐stage aquaculture of this and other economically important species.     

Assay performance during validation of freezing channel catfish Ictalurus punctatus (Rafinesque) infected with a Gram‐negative bacterium

Recovery of bacteria from infected fish during population sampling can be affected by factors including the type of assay, method of specimen preservation and concentration of bacteria present. Consequently, before use in field sampling, methods should be validated. The three objectives of this study were, first, to determine whether a channel catfish Ictalurus punctatus (Rafinesque) fingerling classified as positive for Gram‐negative Edwardsiella ictaluri infection according to bacterial culture before freezing was also classified as positive after freezing, second, to determine how direct culture from the kidney (DIRECT), culture of homogenate (HOMOG) and standard PCR (PCR) agree with bacterial culture in terms of classifying fish as positive or negative and third, to estimate diagnostic sensitivity (dSe) and diagnostic specificity (dSp) for DIRECT, HOMOG and PCR. In fresh and frozen fish, as bacterial concentration decreased, the ability of each assay to detect positive fish also decreased, especially when there were <10⁴ colony‐forming units per gram (CFU g^?1) tissue. HOMOG proved to be the most reliable at correctly classifying catfish, whether they were subclinically or clinically infected. PCR assay was the least reliable. Overall, values for this study population for dSe were 0.66, 0.92 and 0.43, and for dSp were 0.86, 0.91 and 0.95, for DIRECT, HOMOG and PCR respectively.     

Growth Response and Resistance to Streptococcus iniae of Nile Tilapia, Oreochromis niloticus, Fed Diets Containing Distiller's Dried Grains with Solubles

This study was conducted to evaluate the effect of dietary levels of distiller’s dried grains with solubles (DDGS) on growth, body composition, hematology, and resistance of Nile tilapia, Oreochromis niloticus, to Streptococcus iniae challenge. Five isocaloric diets containing DDGS at levels of 0, 10, 20, and 40%, and 40% DDGS + lysine (Diets 1–5) as partial replacements of a combination of soybean meal (SBM) and corn meal (CM) on an equal protein basis were fed to juvenile Nile tilapia (9.41 ± 0.14 g) for 10 wk. Fish fed Diet 4 had the lowest weight gain (WG), feed efficiency ratio, protein efficiency ratio (PER), and whole‐body protein. Supplementation of lysine to the 40% DDGS diet (Diet 5) improved WG and PER. Hematological and immunological parameters were not affected by dietary treatment. There were no significant differences among the average number of days to first mortality after S. iniae challenge and cumulative mortality 14 d postchallenge among fish in various treatments. DDGS can be incorporated in tilapia diet at a level of 20% as a substitute for a combination of SBM and CM without affecting their growth performance, body composition, hematological parameters, immune response, and resistance to S. iniae infection.     

Surgical management of an ethmoid cyst in a horse.

A 2-year old Thoroughbred filly was examined for a 1-month history of persistent nasal discharge. Contrast radiography revealed a circumscribed mass within the right maxillary sinus which extended to the frontal sinus and ethmoid labyrinth. A discrete attachment of the mass to the ethmoid labyrinth was identified at surgery. Surgical removal of the mass eliminated the nasal discharge. On gross examination, the external structure of the mass was similar to a turbinate with a thin bony wall covered by a smooth mucosal membrane. The internal structure of the lesion had a lining membrane with multiple 1-3 cm in diameter fluid filled cystic structures. The histological appearance of the multiloculated structure was similar to the ethmoid labyrinth. This, combined with the single site of attachment to the ethmoid labyrinth, suggested that the cyst was ethmoidal in origin.     

Effect of hypertonic vs isotonic saline solution on responses to sublethal Escherichia coli endotoxemia in horses

Cardiovascular responses to sublethal endotoxin infusion (Escherichia coli, 50 micrograms/ml in lactated Ringer solution at 100 ml/h until pulmonary arterial pressure increased by 10 mm of Hg) were measured 2 times in 5 standing horses. In a 2-period crossover experimental design, horses were either administered hypertonic (2,400 mosm/kg of body weight, IV) or isotonic (300 mosm/kg, IV) NaCl solution after endotoxin challenges. Each solution was administered at a dose of 5 ml/kg (infusion rate, 80 ml/min). Complete data sets (mean arterial, central venous, and pulmonary arterial pressures, pulmonary arterial blood temperature, cardiac output, total peripheral vascular resistance, heart rate, plasma osmolality, plasma concentration of Na, K, Cl, and total protein, blood lactate concentration, and PCV) were collected at 0 (baseline, before endotoxin infusion), 0.25, 1, 1.5, 2, 2.5, 3, 3.5, 4, and 4.5 hours after initiation of the endotoxin infusion. Blood constituents alone were measured at 0.5 hour and cardiovascular variables alone were evaluated at 0.75 hour. By 0.25 hour, endotoxin infusion was completed, a data set was collected, and saline infusion was initiated. By 0.75 hour, saline solutions had been completely administered. Mean (+/- SEM) cardiac output decreased (99.76 +/- 3.66 to 72.7 +/- 2.35 ml/min/kg) and total peripheral resistance (1.0 +/- 0.047 to 1.37 +/- 0.049 mm of Hg/ml/min/kg) and pulmonary arterial pressure (33.4 +/- 0.86 to 58.3 +/- 1.18 mm of Hg) increased for both trials by 0.25 hour after initiation of the endotoxin infusion and prior to fluid administration. For the remainder of the protocol, cardiac output was increased and total peripheral resistance was decreased during the hypertonic, compared with the isotonic, saline trial.(ABSTRACT TRUNCATED AT 250 WORDS)