Effects of Addition of Tissue‐Type Plasminogen Activator in In Vitro Fertilization Medium on Bovine Embryo Development and Quality

Plasminogen activators/Plasmin system plays pivotal role in regulating reproductive functions of mammals. Here, we examined the effects of modification of in vitro fertilization medium (IVF medium) with the addition of tissue‐type plasminogen activator (t‐PA), on bovine embryo development and quality, assessed by quantification of expression of various genes related to metabolism, oxidation, implantation and apoptosis. In addition, plasminogen activator activity (PAA) and plasminogen activator inhibition (PAI) were measured in the spent media. After conventional IVM, 2016 cumulus‐oocyte complexes (COCs) were divided into four groups with modified composition of the IVF medium containing t‐PA and/or its inhibitor epsilon‐aminocaproic acid (control, t‐PA, t‐PA+ε‐ACA, ε‐ACA). Presumptive zygotes were cultured for 8 days in synthetic oviductal fluid (SOF) medium; gene expression studies were carried out on morulae and blastocysts. t‐PA alone significantly suppressed cleavage and blastocyst formation rates, but this effect was neutralized by the addition of ε‐ACA. PAA in the treated group was significantly reduced by ε‐ACA, but without total elimination. Significant differences were detected in the expression of genes related to apoptosis and/or cell cycle arrest (BAX, BCL2L1, KAT2B) between embryos produced in t‐PA‐modified media and controls, giving an overall notion that the inferior developmental competence of treated embryos may be attributed to apoptotic phenomena induced by t‐PA. In conclusion, it appears that excessive t‐PA content in the IVF media, suppresses blastocyst formation rate, possibly due to induction of apoptotic phenomena.     

Ultrastructural analysis of sequential cyprinid herpesvirus 3 morphogenesis in vitro

Cyprinid herpesvirus 3 (CyHV‐3) is an alloherpesvirus, and it is the aetiological agent of koi herpesvirus disease. Although the complex morphogenic stages of the replication cycle of CyHV‐3 were shown to resemble that of other members of the Herpesvirales, detailed analysis of the sequence and timing of these events was not definitively determined. This study describes these features through a time course using cyprinid cell cultures (KF‐1 and CCB) infected with CyHV‐3 (KHV isolate, H361) and analysed by transmission electron microscopy. Rapid viral entry was noted, with high levels of intracellular virus within 1–4 h post‐infection (hpi). Intranuclear capsid assembly, paracrystalline array formation and primary envelopment of capsids occurred within 4 hpi. Between 1 and 3 days post‐infection (dpi), intracytoplasmic secondary envelopment occurred, as well as budding of infectious virions at the plasma membrane. At 5–7 dpi, the cytoplasm contained cytopathic vacuoles, enveloped virions within vesicles, and abundant non‐enveloped capsids; also there was frequent nuclear deformation. Several morphological features are suggestive of inefficient viral assembly, with production of non‐infectious particles, particularly in KF‐1 cells. The timing of this alloherpesvirus morphogenesis is similar to other members of the Herpesvirales, but there may be possible implications of using different cell lines for CyHV‐3 propagation.     

The influence of diel period on fish assemblage in the unstructured littoral of reservoirs

Abstract Diel changes in littoral fish assemblage were studied in four reservoirs in the Czech Republic (Central Europe). The sampling was performed by beach seining in an unstructured littoral zone. Perch, Perca fluviatilis L., roach, Rutilus rutilus (L.), bream, Abramis brama (L.), carp, Cyprinus carpio L., ruffe, Gymnocephalus cernua (L.), pikeperch, Sander lucioperca (L.), eel, Anguilla anguilla (L.) and A. brama × R. rutilus hybrids exhibited higher densities at night. Only bleak, Alburnus alburnus (L.) exhibited higher densities during the day. The number of species was higher in night hauls, and mean body size of roach and bleak was also higher at night. Diel changes in fish densities resulted in the change in species composition between day and night. The study has implication for the sampling design to assess littoral fish assemblages using beach seine netting and recommends night sampling for a representative assessment or sampling during both diel periods for a robust assessment.     

The use of grain protein deviation for identifying wheat cultivars with high grain protein concentration and yield

The relationship between grain protein concentration and grain yield in different cultivars of winter wheat was examined in a series of field experiments carried out over three years, in which 13, 12 and 8 cultivars were studied in each year, respectively. The plants were grown at sites located in Shropshire, west-central England, in years 1 and 2, and at three other locations in eastern England in year 3. Above ground plant samples were collected at an thesis and again at maturity, when they were separated into grain and straw, and analysed for dry matter and N content. Analysis of residuals from regression of grain protein concentration on grain yield (grain protein deviation, GPD) showed that some cultivars had a higher grain protein concentration than was predicted from grain yield alone. It was deduced that the capacity to accumulate a higher grain protein concentration than predicted from grain yield is under genetic control and thus may be improved through breeding. Other factors (weight of N accumulated in the biomass at anthesis, weight of N accumulated in the biomass between anthesis and maturity and the concentration of N remaining in the straw at maturity) were added step-wise into the regression to enable statistical analysis of their relative contributions to grain protein. High GPD may be achieved through increased N accumulation after anthesis, combined with efficient re-translocation of vegetative N reserves. The use of GPD provides a selection criteria in wheat breeding programs to screen for increased grain protein concentration without a concurrent grain yield reduction. This revised version was published online in August 2006 with corrections to the Cover Date.     

Dispersal ability determines the genetic effects of habitat fragmentation in three species of aquatic insect

• 1. The dispersal ability of species and the geographic scale of habitat fragmentation both may influence the extent of gene flow between fragments, but their interactions have rarely been tested, particularly among co‐occurring species.
• 2. Population genetic structures of three species of aquatic insect were compared in streams fragmented by reservoirs and in unfragmented streams in north‐eastern Japan, using 52, 37, and 58 RAPD markers. The three species studied included a strong disperser Cincticostella elongatula (Ephemeroptera: Ephemerellidae), an intermediate disperser Stenopsyche marmorata (Trichoptera: Stenopsychidae), and a weak disperser Hydropsyche orientalis (Trichoptera: Hydropsychidae).
• 3. The patterns of genetic isolation by distance (IBD) supported a priori hypotheses of dispersal ability. The strong disperser (C. elongatula) exhibited significant IBD only at the largest spatial scale studied (among drainages, r=0.50, P<0.01). The intermediate disperser (S. marmorata) showed IBD both within (r=0.22, P<0.01) and among (r=0.45, P<0.01) drainages. The weak disperser (H. orientalis) did not exhibit significant IBD at any scale.
• 4. Pairwise genetic differentiation (θ) indicated that neither the weak disperser nor the strong disperser were genetically differentiated above and below reservoirs when compared with reference sites. This was in contrast to previous results for S. marmorata, for which subpopulations were genetically fragmented across larger (>4.1 km), but not smaller (<2.9 km) reservoirs.
• 5. We suggest that intermediate dispersers, i.e. those at equilibrium between migration and genetic drift within drainages, are more likely to be affected by fragmentation than either strong or weak dispersers. Intermediate dispersers could therefore be used as indicator species in studies aimed at detecting the effects of distance between habitat fragments (e.g. reservoir size) for conservation planning. Copyright © 2010 John Wiley & Sons, Ltd.

     

Enzyme-linked immunosorbent assay (ELISA) for the detection of spring viraemia of carp virus (SVCV) in tissue homogenates of the carp, Cyprinus carpio L.

Abstract. An enzyme-linked immunosorbent assay (ELISA) for the demonstration of the virus of spring viraemia of carp (SVCV) in liver, kidney and spleen homogenates, and in infected cell cultures is described. The sensitivity of the method is 10^2·8–10^3·5 TCID₅₀ 0·lml⁻¹ of the examined fluid. The specificity has been confirmed by the ELISA inhibition test and by results of virological examinations. Contamination with bacteria or fungi of samples taken from dead fish had no effect on the results of ELISA. Specific anti-SVCV sera were used successfully for the production of conjugates for the direct immunoperoxidase and immunofluorescence detection of SVCV in infected cell cultures.     

鲜切生菜供应链温度波动与微生物含量变化

选取北京某优质农产品种植公司的6月中旬、12月下旬2条具有代表性的鲜切生菜供应链为研究对象,对整个链条温度波动规律分别进行为期7d的实地考察测量;并根据调研的温度曲线,在实验室条件下模拟供应链温度,测定鲜切生菜的微生物含量变化.研究结果显示,2条供应链温度都呈现出较大波动性：6月中旬实际供应链温度波动范围为在1.2～24.1℃,12月下旬为-6.1～16.2℃;运输过程温度波动较小,装车卸货等环节温度波动较明显.实验室模拟条件下得到的鲜切生菜微生物含量变化规律与供应链温度波动曲线存在正相关性,相关系数6月中旬0.849 77,12月下旬为0.847 99.     

Recovery of brodifacoum in vomitus following induction of emesis in dogs that had ingested rodenticide bait

AIM: To assess the benefit of inducing emesis in dogs that have ingested rodenticide bait containing brodifacoum (BDF), by determining the amount of BDF in bait recovered from the vomitus relative to the estimated amount consumed.

METHODS: Between 2014 and 2015 samples of vomitus from seven dogs that ingested rodenticide baits containing BDF were submitted by veterinarians in New Zealand. All seven dogs had been given apomorphine by the veterinarian and vomited within 1 hour of ingesting the bait. Some or all of the bait particles were retrieved from each sample and were analysed for concentrations of BDF using HPLC. Based on estimations of the mass of bait consumed, the concentration of BDF stated on the product label, and the estimated mass of bait in the vomitus of each dog, the amount of BDF in the vomited bait was calculated as a percentage of the amount ingested.

RESULTS: For five dogs an estimation of the mass of bait ingested was provided by the submitting veterinarian. For these dogs the estimated percentage of BDF in the bait retrieved from the vomitus was between 10–77%. All dogs were well after discharge but only one dog returned for further testing. This dog had a normal prothrombin time 3 days after ingestion.

CONCLUSIONS AND CLINICAL RELEVANCE: The induction of emesis within 1 hour of ingestion can be a useful tool in reducing the exposure of dogs to a toxic dose of BDF. The BDF was not fully absorbed within 1 hour of ingestion suggesting that the early induction of emesis can remove bait containing BDF before it can be fully absorbed.