An automated, multiplex-tandem PCR platform for the diagnosis of gastrointestinal nematode infections in cattle: An Australian-European validation study.

Detecting the genera and species of gastrointestinal (GI) nematode infections in faecal samples obtained from cattle requires the incubation of faeces ('larval culture') followed by identification of the third-stage larvae that are harvested after 10-14days. Substantial research in the development of PCR-based methods for the rapid and specific identification GI nematodes has been conducted for small ruminants, whilst only few such assays have been developed for cattle. In the present paper we describe the development of an automated, robotic PCR platform for the detection and genus and/or species-specific identification of GI nematodes from bovine faecal samples. This test was then validated using samples from different regions of three countries (Australia, Belgium and Scotland). The PCR platform was found to be highly sensitive and specific for the identification of the important GI nematodes in naturally infected cattle (both estimates >90%). The PCR platform can also estimate the percentage of genera or species present in a mixed-species infection, and was found superior to larval culture in terms of speed (1-2days versus 1-2 weeks for culture), sensitivity and specificity. The PCR was simple to use and the operator requires no knowledge or experience to identify the nematodes present, compared to larval culture where even experienced operators can make substantial errors due to considerable overlap in the published characteristics of key species.

Cattle rabies vaccination--A longitudinal study of rabies antibody titres in an Israeli dairy herd.

In contrast to many regions of the world where rabies is endemic in terrestrial wildlife species, wildlife rabies has been controlled in Israel by oral rabies vaccination programs, but canine rabies is re-emerging in the northern area of the Golan Heights. From 2009 to 2014 there were 208 animal rabies cases in Israel; 96 (46%) were considered introduced primary cases in dogs, triggering 112 secondary cases. One third (37/112) of the secondary cases were in cattle. Rabies vaccination is voluntary for cattle in Israel, except those on public exhibit. Rabies vaccination schedules for cattle vary based on farm practices and perception of risk. In this study 59 cattle from a dairy farm which routinely vaccinates against rabies were assigned into six groups according to age and vaccination histories. Four groups contained adult cows which had received one previous rabies vaccination, one group of adults had received two previous vaccinations, and one group was unvaccinated calves. Serum samples were collected and the cows were vaccinated with a commercial rabies vaccine. Sera were again collected 39 days later and the calf group re-vaccinated and re-sampled 18 days later. Sera were analyzed for the presence of rabies virus neutralizing antibodies using the rapid immunofluorescent antibody test. Cattle with antibody titres ≥ 0.5 IU/ml were considered to be protected against rabies. Twenty-six of 27 adult cattle (96%) vaccinated once at less than five months old did not have protective titres. Sixty percent (6/10) cattle vaccinated once at around six months of age did have adequate titres. Cattle previously vaccinated twice (n=10; 100%) with an 18 month interval between inoculations, had protective titres and protective antibody titres following booster vaccination (n=51; 100%). The anamnestic response of cattle to a killed rabies vaccine was not affected by the time interval between vaccinations, which ranged from 12 to 36 months. These results suggest that calves from vaccinated cows should not be vaccinated before six months old to avoid maternal antibody interference. Whilst most cattle older than six months old will be protected after a single inoculation, a second inoculation ensures a higher antibody levels for improved protection. Cattle receiving an effective priming dose responded well to a booster up to 36 months later. Such results demonstrate the effectiveness of rabies vaccination in cattle and the added value of a second dose to ensure a prolonged immune response against rabies.

The cat flea (Ctenocephalides f. felis) is the dominant flea on domestic dogs and cats in Australian veterinary practices.

This study was undertaken to determine the flea diversity on urban dogs and cats in Australia in 2009-2010. A total of 2530 fleas were recovered from 291 animals (151 dogs, 69 cats and 71 uncategorised dogs or cats) from veterinary clinics across five states of Australia. The majority of specimens were from coastal areas. The cat flea (Ctenocephalides felis felis) was the most frequent flea species identified (98.8%, 2500/2530). The only other flea species identified was the stickfast flea (Echidnophaga gallinacea) from Western Australia. Sequencing of the cytochrome oxidase subunit II mtDNA revealed a single haplotype across Australia within a subset of C. f. felis (n=19). Our study demonstrated dominance and haplotype homogeneity of C. f. felis on dogs and cats. Although Ctenocephalides canis was recovered from a feral fox, it was not identified from the sample of fleas analysed. This suggests that, under current conditions, it is unlikely that foxes are reservoirs of C. canis for domestic dogs or cats residing in coastal Australia, as previously speculated.