Differential diagnosis of dog hookworms based on PCR-RFLP from the ITS region of their rDNA

Species of Ancylostoma infecting dogs and sometimes humans are sympatric in many parts of the world. The establishment of a specific molecular diagnostic tool is important, not only to refine information for epidemiological studies, but also to evaluate the efficacy of vaccine programmes and assist in the development of specific drug treatments. The ITS region from 20 specimens of A. braziliense, collected from three separate geographical areas of Brazil, and from 10 specimens of A. caninum, collected from the same area in Brazil were sequenced and analyzed. Alignment of sequences showed that this gene is highly conserved. The intraspecific polymorphism for both species was less then 1%, whereas the interspecific polymorphism was 6.2, 7.3 and 9.4% between A. ceylanicum and A. braziliense; A. caninum and A. ceylanicum and A. ceylanicum and A. braziliense, respectively. Among the three species it was 12.3%. This revealed the ITS region as highly conserved and consequently a good molecular marker for diagnostic studies. In this work, four restriction enzymes were used in a PCR-RFLP using the ITS region of rDNA, to establish a differential diagnosis which discriminates between three Ancylostoma species, A. braziliense, A. caninum and A. ceylanicum. The best pattern was given by the HinfI enzyme, which produced different fragment sizes for each of the three species. Furthermore, the diagnostic tool differentiates DNA extracted directly from faeces of Ancylostoma-infected dogs.     

Investigation and management of a cluster of cases of equine retained fetal membranes in Highland ponies

Four at-risk Highland ponies on the same premises all retained their fetal membranes in the same breeding season. The ponies were treated with a combination of oxytocin, infusion and distension of the allantochorionic sac with fluid, gentle traction in one case, and supportive therapy. The possible causes of the condition were investigated with clinicopathological tests and analyses of the herbage for its species composition and fungal contamination. No obvious possible causative abnormalities were identified apart from a low total serum calcium level in one case. No fescue grass was found and no pathogenic fungi were detected.     

Seroprevalence of Coxiella burnetii in selected populations of domestic ruminants in Newfoundland

The seroprevalence of Coxiella burnetii among cattle, sheep, and goats in Newfoundland was determined by microimmunofluorescence. Seropositivity to phase II antigen increased in sheep from 3.1% in 1997 to 23.5% in 1999-2000 (P < 0.001). Cows (24%) had antibodies to phase I antigen; goats (15.6%) had antibodies to phase II antigen. Seroprevalence of C. burnetii is increasing among sheep.     

Use of a marker organism in poultry processing to identify sites of cross‐contamination and evaluate possible control measures

1. Nine different sites at a poultry processing plant were selected in the course of a hazard analysis to investigate the degree of microbial cross‐contamination that could occur during processing and the effectiveness of possible control measures.

2. At each site, carcases, equipment or working surfaces were inoculated with a non‐pathogenic strain of nalidixic acid‐resistant Escherichia coli K12; transmission of the organism among carcases being processed was followed qualitatively and, where appropriate, quantitatively.

3. The degree of cross‐contamination and the extent to which it could be controlled by the proposed measures varied from one site to another.     

A survey for Listeria species, motile aeromonads and Yersinia enterocolitica on bovine and ovine carcasses

One hundred ovine and 100 bovine carcasses in two abattoirs were sampled just after dressing for the presence of Yersinia enterocolitica, Listeria monocytogenes and motile aeromonads. Yersinia enterocolitica was not isolated and only two samples were positive for Listeria spp. In both cases, the Listeria species were not normally pathogenic to man. In contrast, motile aeromonads were isolated from 81% of ovine and 35% of bovine carcasses.     

Microbiological survey of five poultry processing plants in the UK

1. Neck skin samples were taken from chickens and turkeys at all the main stages of processing to monitor changes in total viable count (TVC) and counts of coliforms and pseudomonads.

2. Processing reduced TVC by up to 100‐fold. Geometric mean counts after packaging were log₁₀ 4.4 to 5.3 CFU/g whilst corresponding counts of coliforms were 2.7 to 3.8 CFU/g.

3. Increases in mean TVC or coliforms as a result of either defeathering or evisceration did not exceed 0.6 log.

4. Pseudomonads represented only a minor fraction of the initial microflora of the bird and were often reduced by scalding to a figure which could not be detected by direct plating of samples; however, subsequent contamination resulted in means between log₁₀ 2.9 and 4.0 CFU/g for packaged carcases.

5. Although Staphylococcus aureus was readily isolated from defeathering equipment, mean counts from defeathered carcases were always below log₁₀ 3.0 CFU/g.