Dot immunobinding assay method with chlorophyll removal for the detection of southern rice black-streaked dwarf virus.

Southern rice black-streaked dwarf virus (SRBSDV), a new virus from Fiji, has seriously damaged rice crops in southern China and northern Vietnam in recent years. This virus is difficult to diagnose in the early stages of infection, and is very destructive at the late stage. In the present study, a dot immunobinding assay (DIBA) that has a high sensitivity for diagnosing SRBSDV was developed. Two kinds of treatment for the DIBA were evaluated to determine the most effective one for removing chlorophyll interferences via rice extraction. The first included several reagents to remove chlorophyll, namely, the alkaline reagents like magnesium oxide and alumina oxide, the adsorbent reagents like activated carbon and bentonite, as well as the extraction agent acetone. The second and third treatments, which were used to remove chlorophyll in blot membrane-nitrocellulose and polyvinylidene fluoride (PVDF), included several organic solvents containing methanol, ethanol, acetone, ethyl acetate, and diethyl ether. The results showed that activated carbon and methanol yielded the best contrasting purple color for the infected samples by decreasing the chlorophyll content.

The isolation of Kemerovo group orbiviruses and Uukuniemi group viruses of the family bunyaviridae from Ixodes uriae ticks from the Isle of May, Scotland.

Viruses isolated from ticks (Ixodes uriae) from a seabird colony on the Isle of May, Scotland, were shown by complement fixation tests to be related to the Uukuniemi and Kemerovo serogroups. Electron microscopic studies on the Uukuniemi viruses showed them to have a morphology characteristic of the bunyaviridae, and the Kemerovo group to be characteristic of orbiviruses. Separate isolates from the two serogroups were distinguished from each other by neutralization tests.

Reovirus-like agent associated with fatal diarrhea in neonatal pigs.

Large numbers of a reovirus-like agent were visualized with electron microscopy in bacteria-free gut homogenates obtained from piglets with a fatal diarrhea resembling transmissible gastroenteritis. The syndrome, of vomiting, diarrhea, dehydration, and death, was reproduced in piglets artificially infected with these bacteria-free gut homogenates. Reovirus-like particles persisted in serial piglet passage and none was seen in uninfected, asymptomatic controls. Hyperimmune sera (made in recovered piglets) aggregated the reovirus-like particles, as judged by immunoelectron microscopy, and neutralized the infectious agent. The cytoplasm in enterocytes on infected intestinal epithelium fluoresced when this hyperimmune sera was used in an indirect fluorescent antibody test. Feeding cow colostrum or diets containing porcine gamma globulin protected infected piglets. No cytopathogenic effect was noted in infected tissue cultures, nor did this agent affect neonatal guinea pigs, hamsters, mice, and rats. The agent did not agglutinate human O or A erythrocytes.

Studies on rotavirus infection and diarrhoea in young calves.

The occurrence of diarrhoea in calves was monitored during the first three weeks of life. Calves fed amounts of colostrum sufficient to produce serum Ig levels in excess of 30 mg per ml did not develop diarrhoea, whereas calves fed less colostrum did. Rotaviruses and mycoplasma-like particles were observed in the faeces of calves with and without diarrhoea. The epidemiology of rotavirus infection in calves is discussed.

Viral enteritis of calves.

Calf diarrhoea is an important cause of economic loss. The aetiology of the disease is considered with particular reference to the role of viruses. Although many microbial organisms have been associated with the disease, there is doubt concerning their true role as causative agents. Two viruses, neonatal calf diarrhoea reovirus-life agent and calf coronavirus, have been discovered recently which appear to play an important role in many cases of diarrhoea and the evidence for considering them to be primary causes of the disease is discussed.

Concentration of enteric viruses from water with lettuce extract.

A method for recovering enteroviruses, adenovirus, and reovirus from water with lettuce extract is described. Lettuce extract at pH 8.5 was added to the sample and the pH was reduced stepwise with hydrochloric acid to 4.0 to 4.5. The flocculent lettuce-extract particles, and adsorbed virus, were readily removed from solution by low-speed centrifugation. Electron microscopy suggests that, under conditions suitable for adsorption, virus particles are coated with the lettuce-extract colloid.