No abstract

Tilapia are an important group of farmed fish that serve as a significant protein source worldwide. In recent years, substantial mortality of wild tilapia has been observed in the Sea of Galilee and in commercial ponds in Israel and Ecuador. We have identified the etiological agent of these mass die-offs as a novel orthomyxo-like virus and named it tilapia lake virus (TiLV). Here, we provide the conditions for efficient isolation, culturing, and quantification of the virus, including the use of susceptible fish cell lines. Moreover, we describe a sensitive nested reverse transcription-PCR (RT-PCR) assay allowing the rapid detection of TiLV in fish organs. This assay revealed, for the first time to our knowledge, the presence of TiLV in diseased Colombian tilapia, indicating a wider distribution of this emerging pathogen and stressing the risk that TiLV poses for the global tilapia industry. Overall, the described procedures should provide the tilapia aquaculture industry with important tools for the detection and containment of this pathogen.

Using transmission electron microscopy (TEM), the presented work expands on the ultrastructural findings of an earlier report on "syncytial hepatitis," a novel disease of tilapia (SHT). Briefly, TEM confirmed the presence of an orthomyxovirus-like virus within the diseased hepatocytes but not within the endothelium. This was supported by observing extracellular and intracellular (mostly intraendosomal), 60-100 nm round virions with a trilaminar capsid containing up to 7 electron-dense aggregates. Other patterns noted included enveloped or filamentous virions and virion-containing cytoplasmic membrane folds, suggestive of endocytosis. Patterns atypical for orthymyxovirus included the formation of syncytia and the presence of virions within the perinuclear cisternae (suspected to be the Golgi apparatus). The ultrastructural morphology of SHT-associated virions is similar to that previously reported for tilapia lake virus (TiLV). A genetic homology was investigated using the available reverse transcriptase polymerase chain reaction (RT-PCR) probes for TiLV and comparing clinically sick with clinically normal fish and negative controls. By RT-PCR analysis, viral nucleic acid was detected only in diseased fish. Taken together, these findings strongly suggest that a virus is causally associated with SHT, that this virus shares ultrastructural features with orthomyxoviruses, and it presents with partial genetic homology with TiLV (190 nucleotides).

We studied risk factors and characteristics of canine transmissible venereal tumours (TVTs) in Grenada. We abstracted data for 38 TVT cases and 114 TVT-free dogs submitted to a veterinary diagnostic laboratory between 2003 and 2006. Occurrence profiles, odds ratios (ORs), and logistic regression models for TVT were determined using a significance level of alpha = 0.05. TVT was found in 20 (52.6%) female and 18 (47.4%) male dogs. Of the TVT cases, 32 (84.2%) were between 1 and 7 years old, 20 (52.6%) were mixed breeds of dogs, 14 (36.8%) were Grenadian pothounds, while 4 (10.6%) were pure-bred dogs. Characteristic TVT lesions were genital growths [OR = 96.7; 95% CI (27,461), P < 0.001], genital bleeding [OR = 12.7; 95% CI (4.6, 39.2), P < 0.001] and secondary inflammation of TVT lesion [OR = 4.3; 95% CI (2, 10), P < 0.001]. Extragenital TVT lesions were observed in 23% (9/38) of dogs. An increased risk for TVT was associated with age as adult (1-7 years) dogs [OR = 12; 95% CI (1.6, 94), P < 0.001] and status as a Grenadian pothound [OR = 8.6; 95% CI (3, 25), P < 0.001]. Clinicians should educate dog owners about increased risk of TVT for Grenadian pothounds and consider TVT as a possibility for some extragenital tumours.

Here, we report the complete coding sequences of tilapia lake virus (TiLV) associated with syncytial hepatitis of tilapia (SHT). The TiLV strain was sequenced from the liver RNA extract of a moribund Nile tilapia (Oreochromis niloticus) fingerling from an Ecuadorian aquaculture facility in 2012.