Hemagglutinating activity of Gallibacterium strains.

In the present study, the hemagglutinating activity of seven reference strains, and nine Mexican and three Danish field isolates, of Gallibacterium was investigated by using fresh erythrocytes of 19 different types including chicken (broiler, rooster, layer hen), turkey, pigeon, quail, duck, Harris's hawk (Parabuteo unicinctus), house finch (Carpodacus mexicanus), cow, sheep, horse, dog, rabbit, pig, and human (groups A, B, AB, and O; Rh+). Agglutination was observed for broiler chicken, layer hen, quail, rabbit, and pig erythrocytes with a subset of Gallibacterium strains, whereas most tested strains agglutinated rabbit erythrocytes. Transmission electron microscopic examination of a hemagglutinating strain demonstrated a close interaction between the bacterial and erythrocyte surfaces. The results indicate that some Gallibacterium strains are able to agglutinate avian or mammalian erythrocytes, or both. However, the mechanisms enabling hemagglutination are not known and will be addressed in future studies.

Development and validation of a short-time cell culture and multiplex reverse transcriptase polymerase chain reaction assay for infectious pancreatic necrosis virus in Mexican farm-sampled rainbow trout.

The infectious pancreatic necrosis virus (IPNV) affects several species of freshwater and marine fish. In Mexico, IPNV has an important impact on farming of rainbow trout Oncorhynchus mykiss; however, IPNV distribution in Mexico is unclear. The diagnosis of IPNV is laborious; usually it is based on isolation tests in cell culture followed by immunological identification using techniques of serum neutralization, immunofluorescence, or enzyme-linked immunosorbent assay. It has recently been demonstrated that reverse transcriptase polymerase chain reaction (RT-PCR) is an adequate method for the detection of aquatic birnaviruses. However, its diagnostic use is still limited because very low titers of viable virus cannot be easily detected. In this study, a combination of short-time cell culture and multiplex RT-PCR was established for the diagnosis of IPNV in rainbow trout obtained from farms in the state of Mexico. Three primer sets were used in a single reaction in the multiplex RT-PCR to increase the probability of identifying all serotypes of IPNV serogroup A as well as to help prevent a false-negative result. This approach was able to identify samples with an IPNV concentration of just 0.01 tissue culture infective dose with 50% endpoint (TCID50)/mL, and it identified more infected fish than RT-PCR alone or first-passage cell culture alone. Moreover, this technique made the same identifications as second-passage cell culture but in approximately 30% of the time needed for second-passage cell culture. Consequently, the time and cost efficiency of IPNV diagnosis were greatly reduced.

The T963C mutation of TP53 gene does not participate in the clonal origin of canine TVT.

In dogs, the canine transmissible venereal tumor (CTVT) is the only neoplasm which is not produced by neoplastic transformation of normal cells; the tumor is transmitted from the affected dog to healthy dogs by implantation of one or various clones of cancer cells. Thus, the CTVT of dogs analyzed in various countries reveals similar genetic characteristics and consequently CTVT is considered to have a clonal origin. The CTVTs obtained from dogs in Korea showed the T963C mutation on TP53 gene; this mutation was thought to be a molecular alteration which participates in the origin of the ancestral clone, CTVT. Nonetheless, this supposed mutation has not been identified in other studies which were carried out for the purpose of clarifying the clonal origin of CTVT. Thus we have considered it important to identify the role of the T963C mutation of the TP53 gene in the clonal origin of CTVT in dogs. Consequently the region which includes the mutation of the TP53 gene in twenty samples of CTVT obtained from various canine breeds was PCR amplified and afterwards its sequence of nucleotides was determined. We conclude that this mutation did not participate in the clonal origin of the tumor, but was acquired at a later stage.

Identification of a genetic variant of canine distemper virus from clinical cases in two vaccinated dogs in Mexico.

Canine distemper virus (CDV) is a highly contagious viral pathogen of worldwide distribution that can cause lethal disease in dogs and other mammals. Genetic diversity is found among reference strains and isolates of CDV, mainly in the haemagglutinin protein (H), fusion protein (F) and nucleoprotein (N), and this may be associated with the increasing incidence of distemper in dogs. CDV was identified by RT-PCR in serum samples taken from two clinically diseased, previously vaccinated Mexican dogs. Subsequently, in both samples, a fragment of the CDV N gene was sequenced revealing a 100% identity between nucleotide sequences. However, the sequence obtained was different to that found in virus strains used in vaccines and in isolates reported elsewhere, but was closely related to A75/17, 1127/Gi95, and 2495/Gi95 sequences from USA and Germany, and clustered with 1127/Gi95 and 2495/Gi95 strains. The results suggest that a novel CDV lineage may be present in Mexico.