Rapid detection of bovine herpesvirus 1 in the semen of infected bulls by a nested polymerase chain reaction assay.

A nested polymerase chain reaction (PCR) assay was developed for the detection of bovine herpesvirus 1 (BHV-1) in bovine semen and compared with the virus isolation method. When extended semen, commonly used in the bovine artificial insemination industry, was inoculated with BHV-1, the PCR assay detected BHV-1 DNA in semen inoculated at 0.25-2.5 TCID50 per 0.5 mL. In contrast, the lower limit of detection for virus isolation was 250 TCID50 of BHV-1 inoculated in 0.5 mL of extended semen. These methods were also used to detect BHV-1 in the semen of four bulls which were experimentally infected with BHV-1. All infected bulls demonstrated balanitis at 3 d post-inoculation (DPI) and severe balanoposthitis at 4 DPI. BHV-1 was detected in raw semen by virus isolation and PCR at 2 DPI, before balanitis was evident. For virus isolation, the last day that BHV-1 was detected during primary infection was 7 DPI for two bulls and 9 and 11 DPI for the other two bulls. In contrast, PCR detected BHV-1 in the bulls' semen until 14 or 18 DPI. For individual animals, PCR detected BHV-1 during primary infection for at least 1-10 d longer than virus isolation. Reactivation of BHV-1 from latency without the presence of visible lesions was promoted twice by two series of 5 d dexamethasone injections. For the first series of dexamethasone treatments, a positive virus isolation result was obtained on the 5th d of treatment for only one bull. In contrast, two bulls demonstrated evidence of viral reactivation on this day by PCR. All bulls shed BHV-1 in semen on d 4 after dexamethasone treatment, as evidenced by positive virus isolation and PCR results. One bull was still PCR positive 13 d later. For the second series of dexamethasone treatments, a small amount of virus was isolated from semen collected on d 3 or 4 after treatment for two bulls but not from the other two bulls. In contrast, semen samples from all bulls were PCR positive for either or both of these 2 d. In total, from 80 semen samples, 45 were PCR positive and 26 were virus isolation positive. Thus, the PCR assay detected BHV-1 shedding in bulls earlier, more often, and for a longer duration, than did the virus isolation method.     

Bat rabies in british columbia 1971-1985

Rabies virus was demonstrated in 99 of 1154 bats submitted from British Columbia between 1971 and 1985. Rabies was diagnosed in seven species including big brown bats (Eptesicus fuscus), the latter accounting for 51% of all positive cases. Colonial species represented 92.9% of all identified bats and 87.7% of all rabid cases. Most bats were submitted from the more densely populated areas of the province, and submissions and positive cases both peaked in the month of August. Daytime activity and inability to fly were the most common behaviors reported in rabid bats.     

Rabies in Western Canada 1978-1984

The results of laboratory examination of 22,503 specimens for the diagnosis of rabies by the standard fluorescent antibody and mouse inoculation tests over a seven year period are presented. Specimens were received from British Columbia, Alberta, Saskatchewan, the Yukon and Northwest Territories. Of the 1,445 positive cases, 10.50% involved human contact. The main reservoirs of rabies were skunks, bats and foxes. The reliability of the fluorescent antibody test for the diagnosis of rabies was reaffirmed by agreement with the mouse inoculation test in over 99% of cases.     

Cortex is driven by weak but synchronously active thalamocortical synapses

Sensory stimuli reach the brain via the thalamocortical projection, a group of axons thought to be among the most powerful in the neocortex. Surprisingly, these axons account for only approximately 15% of synapses onto cortical neurons. The thalamocortical pathway might thus achieve its effectiveness via high-efficacy thalamocortical synapses or via amplification within cortical layer 4. In rat somatosensory cortex, we measured in vivo the excitatory postsynaptic potential evoked by a single synaptic connection and found that thalamocortical synapses have low efficacy. Convergent inputs, however, are both numerous and synchronous, and intracortical amplification is not required. Our results suggest a mechanism of cortical activation by which thalamic input alone can drive cortex.     

The food contaminant fumonisin B1 reduces the maturation of porcine CD11R1+ intestinal antigen presenting cells and antigen-specific immune responses, leading to a prolonged intestinal ETEC infection

Consumption of food or feed contaminated with fumonisin B₁ (FB₁), a mycotoxin produced by Fusarium verticillioides, can lead to disease in humans and animals. The present study was conducted to examine the effect of FB₁ intake on the intestinal immune system. Piglets were used as a target and as a model species for humans since their gastro-intestinal tract is very similar. The animals were orally exposed to a low dose of FB₁ (1 mg/kg body weight FB₁) for 10 days which did not result in clinical signs. However, when compared to non-exposed animals, FB₁-exposed animals showed a longer shedding of F4⁺ enterotoxigenic Escherichia coli (ETEC) following infection and a lower induction of the antigen-specific immune response following oral immunization. Further analyses to elucidate the mechanisms behind these observations revealed a reduced intestinal expression of IL-12p40, an impaired function of intestinal antigen presenting cells (APC), with decreased upregulation of Major Histocompatibility Complex Class II molecule (MHC-II) and reduced T cell stimulatory capacity upon stimulation. Taken together, these results indicate an FB₁-mediated reduction of in vivo APC maturation.     

Effect of food matrix and pH on the volatilization of bases (TVB) in packed north Atlantic gray shrimp (Crangon crangon): volatile bases in MAP fishery products

The total volatile basic nitrogen fraction (TVB-N) is often used as a quality parameter in the fish industry to assess spoilage. This parameter often leads to discussions between producers and retailers when it comes to defining clear limits of acceptability for modified atmosphere (MA) packed fish and fishery products. Suggested product limits (mg N/100 g fish) do not always correlate with the presence of off-odors. Gray shrimp are an economic valuable, very perishable niche product, where the TVB-N fraction plays an important role considering its shelf life. This research focuses on the effect of a shrimp matrix and its pH on the volatilization of these formed bases, revealing the relationship between concentrations in the fishery product and the concentrations of these bases present in the headspace of the packed product. Especially, the pH of the product, which is lowered when fishery products are packed under a carbon dioxide enriched atmosphere, appeared to have an immense effect on the volatilization of these bases. The effect of the fish matrix itself is established by means of calculated equilibration constants (dimensionless) being 2.13×10(-4)±0.38×10(-4) for trimethylamine, 6.34×10(-5)±1.71×10(-5) for dimethylamine, and 2.58×10(-5)±0.49×10(-5) for ammonia. Comparison of these constants with the equilibration constants of an aqueous solution indicated the retention of these bases in the product. This article provides not only the important insights for the interpretation of TVB-N values in modified atmosphere packaged gray shrimp but also the methodology to extend these findings to other fish and fishery products.     

Effect of the spray application technique on the deposition of entomopathogenic nematodes in vegetables

BACKGROUND: The present study compared entomopathogenic nematode delivery at the base of savoy cabbage and cauliflower, at the lower side of savoy cabbage and cauliflower leaves and in leek stems and the ground deposition using a five‐nozzle spray boom equipped with an ISO 08 flat fan, an air induction flat fan and Twinjet spray nozzles. Additionally, an air support system and a row application system were evaluated. RESULTS: Approximately 40% of the applied nematodes did not reach the foot of the cabbage plants. The use of an air support system or a row application system improved nematode deposition at the savoy cabbage base. Relative nematode deposition on the lower side of savoy cabbage leaves was 27.20%, while only 2.64% of the applied nematodes reached the lower side of cauliflower leaves. After spraying leek with a standard boom, a low relative nematode deposition (26.64%) was measured in the leek stem. Nozzle type affected the distribution of nematodes in droplet spots. CONCLUSION: Nozzle type has a minor effect on the number of entomopathogenic nematodes delivered on difficult‐to‐reach targets. The use of modified spray application techniques directing the spray to the target site are necessary to increase the chances of contact of entomopathogenic nematodes with their target. Copyright © 2011 Society of Chemical Industry     

Occurrence of Cucurbit Yellow Stunting Disorder Virus (CYSDV) and Beet Pseudo-yellows Virus in Cucurbit Crops in Spain and Transmission of CYSDV by Two Biotypes of Bemisia tabaci

The relative occurrence in Spain of two whitefly transmitted closteroviruses causing similar yellowing diseases in melon and cucumber greenhouse crops was studied. Based on a RT-PCR assay, a 1994–1997 survey of Spanish greenhouses showed that the recently described Bemisia tabaci-transmitted cucurbit yellow stunting disorder virus (CYSDV) has displaced the Trialeurodes vaporariorum-transmitted beet pseudo-yellows virus (BPYV), a virus that was present in the area since the late 1970s. The CYSDV transmission rates by each of the two biotypes of B. tabaci present in Spain were compared. The results showed that the ubiquitous B biotype and the resident Q biotype (found in Spain and Portugal) were able to transmit CYSDV with similar efficiency.