Development of multi-rotor localised surveillance using multi-spectral sensors for plant biosecurity

This report describes a proof of concept for multi-rotor localised surveillance using a multi-spectral sensor for plant biosecurity applications. A literature review was conducted on previous applications using airborne multispectral imaging for plant biosecurity purposes. A ready built platform was purchased and modified in order to fit and provide suitable clearance for a Tetracam Mini-MCA multispectral camera. The appropriate risk management documents were developed allowing the platform and the multi-spectral camera to be tested extensively. However, due to technical difficulties with the platform the Mini- MCA was not mounted to the platform. Once a suitable platform is developed, future extensions can be conducted into the suitability of the Mini-MCA for airborne surveillance of Australian crops.

Ecology of uncultivated oscillospira species in the rumen of cattle, sheep, and reindeer as assessed by microscopy and molecular approaches

The ecology of the uncultured, but large and morphologically conspicuous, rumen bacterium Oscillospira spp. was studied. Oscillospira-specific 16S rRNA gene sequences were detected in North American domestic cattle, sheep from Australia and Japan, and Norwegian reindeer. Phylogenetic analysis of the sequences obtained allowed definition of three operational taxonomic units within the Oscillospira clade. Consistent with this genetic diversity, we observed atypical smaller morphotypes by using an Oscillospira-specific fluorescence in situ hybridization probe. Despite the visual disappearance of typical large Oscillospira morphotypes, the presence of Oscillospira spp. was still detected by Oscillospira-specific PCR in the rumen of cattle and sheep. These observations suggest the broad presence of Oscillospira species in various rumen ecosystems with the level, and most likely the morphological form, dependent on diet. An ecological analysis based on enumeration of the morphologically conspicuous, large-septate form confirms that the highest counts are associated with the feeding of fresh forage diets to cattle and sheep and in two different subspecies of reindeer investigated.

Toxoplasmosis in Indo-Pacific humpbacked dolphins (Sousa chinensis) from Queensland

Objective: To describe the clinical signs, gross pathology, serology, bacteriology, histopathology, electron microscopy and immunohistochemistry findings associated with toxoplasmosis in four Indo-Pacific humpbacked dolphins (Sousa chinensis) that stranded in Queensland in 2000 and 2001. Design: Clinical assessment, gross necropsy, and laboratory examinations. Procedure: Necropsies were performed on four S chinensis to determine cause of death. Laboratory tests including serology, bacteriology, histopathology and transmission electron microscopy were done on the four dolphins. lmmunohistochemistry was done on the brain, heart, liver, lung, spleen and adrenal gland from various dolphins to detect Toxoplasma gondii antigens. Results: Necropsies showed all of four S chinensis that stranded in Queensland in 2000 and 2001 had evidence of predatory shark attack and three were extremely emaciated. Histopathological examinations showed all four dolphins had toxoplasmosis with tissue cysts resembling T gondii in the brain. Tachyzoite stages of T gondii were detected in the lungs, heart, liver, spleen and adrenal gland, variously of all four dolphins. Electron microscopy studies and immunohistochemistry confirmed the tissues cysts were those of Tgondii. All four dolphins also had intercurrent disease including pneumonia, three had peritonitis and one had pancreatitis. Conclusion: Four S chinensis necropsied in Queensland in 2000 and 2001 were found to be infected with toxoplasmosis. It is uncertain how these dolphins became infected and further studies are needed to determine how S chinensis acquire toxoplasmosis. All four dolphins stranded after periods of heavy rainfall, and coastal freshwater runoff may be a risk factor for T gondii infection in S chinensis. This disease should be of concern to wildlife managers since S chinensis is a rare species and its numbers appear to be declining.

Use of Petrifilm(R) 3M to assess coliform numbers on lamb carcasses.

Petrifilm(R) (6410) was used directly on lamb carcasses to enumerate coliforms. 10 sites on 30 carcasses were sampled at each of 4 separate meat processing establishments (works). Coliform counts obtained by this technique were statistically analysed using analysis of variance (ANOVA) to select the optimum sampling sites on the carcass and to assess contamination of the carcass by gut flora at a particular establishment. There was a large variation between sites and between works. In general, works 3 and 4 produced cleaner carcasses than works 2, which in turn was cleaner than works 1. Works 1, 2 and 4 used conventional dressing techniques and works 3 used the inverted dressing method, therefore, the coliform counts found at works 3 and 4 are achievable regardless of dressing technique. Coliform bacteria were most concentrated around the posterior pelvic rim and less prevalent at the carcass extremities. The posterior pelvic rim (sites 3 and 4) had higher (P < 0.05) coliform counts than the exterior ventral flank area (sites 5, 6, 7 and 8), which in turn had higher (P < 0.05) counts than the proximal hind and proximal fore limbs (sites 1, 2, 9 and 10) across all works. With in-line routine testing it is recommended that the majority of carcasses sampled should give coliform counts of <50 cfu/20 cm2 for sites 4 and 8. Reprinted with permission from Journal of Food Protection. Copyright held by the International Association of Food Protection, Des Moines, Iowa, USA. Authors affifiation. J.A.Guthrie & K.J.Dunlop International Food Institute of Queensland, Department of Primary Industries, Rockhampton and G.A.Saunders Veterinary Public Health Division, Livestock and Meat Authority of Queensland, Emerald.

Composition of chemical attractants affects trap catches of the Australian sheep blowfly, lucilia cuprina, and other blowflies

Numbers of Lucilia cuprina (Australian sheep blowfly), Chrysomya spp., and Calliphora spp. blowflies caught on sticky traps baited with various synthetic attractants or a standard liver/sodium sulfide attractant in western Queensland were recorded. Numbers of each genus collected were influenced by the composition of the chemical attractants. Attractant mixtures based on 2-mercaptoethanol, indole, butanoic/pentanoic acid, and a sodium sulfide solution gave 5- to 20-fold higher L. cuprina catches than the liver standard. These blends attracted similar numbers of Chrysomya spp. (0.85–2.7× ) and fewer Calliphora spp. (0.02–0.2× ) compared to the liver standard. These synthetic attractants were more effective and selective for L. cuprina than the standard liver/sodium sulfide attractant, and they can be packaged in controlled-release dispensers to generate constant, prolonged release of the attractant.

Coriza infecciosa: revisión de métodos de diagnóstico y vacunas

La coriza infecciosa es una enfermedad respiratoria aguda de las gallinas domesti- cas causada por la bacteria Haemophilus parugallinarum. Excepcionalmente pueden enfermarse tambien los faisanes y gallinas de Guinea. El H. paragallinarum infecta al ave por via respiratoria y luego de un cor- to periodo de incubation, que varia entre 1 a 3 dias, produce una enfermedad que se manifiesta por inflamacion catarral de los senos paranasales. Este cuadro puede estar asociado a inflamacion de los barbillones, conjuntivitis o queratitis. Los casos de neu- nionia y aerosaculitis son menos frecuentes pero tambien suelen ocurrir en las infeccio- nes puras por estos hemofilos. En las gallinas en produccion causa alta morbilidad, baja o nula mortalidad y una importante perdida en la produccion de huevos, la que generalmente oscila entre 10% y 40%. En pollos parrilleros puede cau- sar un cuadro descrito como «cabeza hin- chada» y ocasionalmente tambien producir septicemia y muerte (48). Esta bacteria ge- neralmente se asocia con otros agentes bacterianos, viricos o parasitarios y cuan- do esto ocurre se agrava el curso de la en- fermedad. Entre los agentes bacterianos mas comunes deben mencionarse los mycoplasinas y las pasteurelas. Cuando H . paragallinarum se asocia con otros agentes esta enfermedad se denomina .«coriza infec- ciosa complicada» (48). En esta recopilacion se aportaran deta- lles sobre la clasificacion, identificacion y serotipificacion del agente causal. Tambien se resumira la informacion disponible sobre nuevos metodos de diagnostico y programas de vacunacion para prevenir esta enferme-dad. A lo largo de esta revision se hara re-ferencia a los hemofilos aviarios que, para el proposito de este trabajo, seran definidos como organisnios gram negativos aislados de aves y que necesariamente requieren factores de crecimiento in vitro. Los dos factores que pueden ser requeridos por los hemofilos para su crecimiento in vitro son hemina (factor X) y/o nicotin-adenin-dinucleirtido (NAD o factor V).

Using simulation techniques to investigate methods to determine resistance of helminths to anthelmintic treatment

The widespread and increasing resistance of internal parasites to anthelmintic control is a serious problem for the Australian sheep and wool industry. As part of control programmes, laboratories use the Faecal Egg Count Reduction Test (FECRT) to determine resistance to anthelmintics. It is important to have confidence in the measure of resistance, not only for the producer planning a drenching programme but also for companies investigating the efficacy of their products. The determination of resistance and corresponding confidence limits as given in anthelmintic efficacy guidelines of the Standing Committee on Agriculture (SCA) is based on a number of assumptions. This study evaluated the appropriateness of these assumptions for typical data and compared the effectiveness of the standard FECRT procedure with the effectiveness of alternative procedures. Several sets of historical experimental data from sheep and goats were analysed to determine that a negative binomial distribution was a more appropriate distribution to describe pre-treatment helminth egg counts in faeces than a normal distribution. Simulated egg counts for control animals were generated stochastically from negative binomial distributions and those for treated animals from negative binomial and binomial distributions. Three methods for determining resistance when percent reduction is based on arithmetic means were applied. The first was that advocated in the SCA guidelines, the second similar to the first but basing the variance estimates on negative binomial distributions, and the third using Wadley’s method with the distribution of the response variate assumed negative binomial and a logit link transformation. These were also compared with a fourth method recommended by the International Co-operation on Harmonisation of Technical Requirements for Registration of Veterinary Medicinal Products (VICH) programme, in which percent reduction is based on the geometric means. A wide selection of parameters was investigated and for each set 1000 simulations run. Percent reduction and confidence limits were then calculated for the methods, together with the number of times in each set of 1000 simulations the theoretical percent reduction fell within the estimated confidence limits and the number of times resistance would have been said to occur. These simulations provide the basis for setting conditions under which the methods could be recommended. The authors show that given the distribution of helminth egg counts found in Queensland flocks, the method based on arithmetic not geometric means should be used and suggest that resistance be redefined as occurring when the upper level of percent reduction is less than 95%. At least ten animals per group are required in most circumstances, though even 20 may be insufficient where effectiveness of the product is close to the cut off point for defining resistance.

A lack of predatory interaction between rumen ciliate protozoa and Shiga-toxin producing Escherichia coli

Aims: To investigate interactions between rumen protozoa and Shiga toxin-producing Escherichia coli (STEC) and to ascertain whether it is likely that rumen protozoa act as ruminant hosts for STEC. Methods and Results: The presence of stx genes in different microbial fractions recovered from cattle and sheep rumen contents and faeces was examined using PCR. In animals shedding faecal STEC, stx genes were not detected in the rumen bacterial or rumen protozoal fractions. Direct interactions between ruminal protozoa and STEC were investigated by in vitro co-incubation. Rumen protozoa did not appear to ingest STEC, a STEC lysogen or non-STEC E. coli populations when co-incubated. Conclusions: The ruminal environment is unlikely to be a preferred habitat for STEC. Bacterial grazing by rumen protozoa appears to have little, if any, effect on STEC populations. Significance and Impact of the Study: This study indicates that ruminal protozoa are unlikely to be a major factor in the survival of STEC in ruminants. They appear as neither a host that protects STEC from the ruminal environment nor a predator that might reduce STEC numbers.

Cross-protection study of the nine serovars of Haemophilus paragallinarum in the Kume haemagglutinin scheme

The cross-protection and haemagglutination-inhibition antibodies present in chickens vaccinated with one of the nine currently recognized Kume haemagglutinin serovars of Haemophilus paragallinarum were investigated. The results confirmed the widely accepted dogma that serogroups A, B, and C represent three distinct immunovars. Within Kume serogroup A, there was generally good cross-protection among all four serovars. However, within Kume serogroup C, there was evidence of a reduced level of cross protection between some of the four serovars. The haemagglutination-inhibition antibody levels generally showed the same trend as with the cross-protection results. This study suggests that some apparent field failures of infectious coryza vaccines may be due to a lack of cross-protection between the vaccine strains and the field strains. Our results will help guide the selection of strains for inclusion in infectious coryza vaccines.

Rejection by pigs of mouldy grain containing deoxynivalenol

Weaner pigs on a farm near Beaudesert in south eastern Queensland refused to eat feed comprised largely of wheat and barley. Older pigs consumed small amounts and some prepubertal gilts subsequently displayed enlarged and reddened vulvas. Wheat, barley and triticale were grown on the farm during 1983, which was unusually and persistently wet. The wheat and triticale were harvested and stored for about 3 weeks with moisture contents above 14% before being fed. Samples of the wheat and triticale contained pale pink grains, which can indicate infection by the fungus Fusariurn grarninearurn Schw. On analysis 2 mycotoxins known to be produced by F. graminearurn were detected, deoxynivalenol (vomitoxin) which causes feed refusal and vomiting, and zearalenone which causes oestrogenic effects. Concentrations of deoxynivalenol in the wheat, triticale and barley were 34, 10, and <0.1 mg/kg respectively. Concentrations of zearalenone were 6.2, 2.8 and 0.1 mg/kg respectively. Subsequently, F. grarninearurn was isolated from grains and crop residues. Although the wet weather contributed to F. grarninearurn infection of the crops before harvest, most of the toxins probably developed during storage.

Zearalenone intoxication of pigs

Mycotoxicosis due to ingestion of zearalenone was detected on 2 pig farms on the Atherton Tableland in northern Queensland. In one herd of 200 pigs, this resulted from feeding maize which had been stored with a high moisture content. In the other herd of 1400 pigs, it resulted from feeding sorghum grain which was rain affected before harvest. Concentrations of zearalenone in the feeds ranged up to 8 mg/kg. Most prepubertal gilts in the herds displayed enlarged teats and signs of oestrus such as having red, swollen vulvas. In several cases both rectal and vaginal prolapses occurred. On one of the farms, 25 pigs died as a direct result of prolapses. Autopsy of a 3-monthold gilt revealed apparently enlarged ovaries and uterine horns. Sows and boars seemed to be unaffected. Four gilts failed to conceive following mating during the period of zearalenone ingestion, but apart from this and the deaths from prolapses, production of the herds appeared ti be unaffected.

Field cases of aflatoxicosis in pigs

Five cases of aflatoxicosis in pigs in southern Queensland are described. One peracute case where aflatoxin concentrations of up to 5000pg aflatoxin B,/kg were demonstrated in stomach contents was presumed to be caused by consumption of mouldy bread. High levels of toxins were also present in the livers. Two cases of acute toxicity were caused by feeding mouldy peanut screenings containing 22000~9 aflatoxin B,/kg. One case of subacute and one of chronic toxicity were caused by sorghum grain based rations with lower aflatoxin levels (4640 and 255 pg/kg). Peracute toxicity caused collapse and deaths within several hours, acute toxicity caused deaths within 12 h and with subacute toxicity deaths occured after 3 weeks on a toxic ration. Anorexia and ill thrift affecting only growing animals were seen with chronic toxicity. Extensive centrilobular liver necrosis and haemorrhage occured with peracute toxicity and in cases of acute poisoning there was hepatic centrilobular cellular infiltration, hepatocyte swelling and bile stasis. With subacute toxicity hepatocyte vacuolation together with bile stasis and bile ductule hyperplasia were seen.

Observations on the influence of repeated sampling procedures on temperament changes of weaner cattle

Experimental cattle are often restrained for repeated blood collection and faecal sampling and may baulk at entering the crush, possibly from learning that crush entry is followed by an unpleasant experience. We asked whether repeated sampling affects temperament. One measure of temperament is flight speed, which is the time, measured electronically, for an animal to cover a set distance on release from a weighing crate (Burrow et al. 1988). 22nd Biennial Conference.

Phenotypic and genotypic characterisation of Pasteurella multocida isolated from pigs at slaughter in New Zealand

AIMS: To examine pigs at slaughter in New Zealand for the presence of Pasteurella multocida, and to determine for isolates, their biochemical profi les, somatic and capsular types, and the presence or absence of the HSB and toxA genes, associated with haemorrhagic septicaemia (HS) and progressive atrophic rhinitis (PAR), respectively. METHODS: Swabs from 173 lungs, 158 palatine tonsils and 82 nasal passages of pigs at two abattoirs in New Zealand were cultured for P. multocida using conventional techniques, and isolated colonies were subjected to biochemical tests for identi- fi cation of biovars. Somatic serotyping was conducted using an agar gel immunodiffusion (AGID) test. Polymerase chain reaction (PCR) assays were used to confi rm phenotypic identifi cation of colonies using species-specifi c primers, capsule type using serogroup-specifi c primers and multiplex PCR, and to test for the presence of HSB and toxA genes. RESULTS: Pasteurella multocida was isolated from 11/173 (6.4%) lung, 32/158 (20.2%) palatine tonsil and 5/82 (6.1 %) nasal swab samples, a total of 48 isolates from 413 samples (11.6%). Isolation rates per farm ranged from 1–53% of tissue samples collected from pigs 5–6 months of age. On phenotypic characterisation, isolates were allocated to seven main biovars, viz 1, 2, 3, 5, 9, 12, and a dulcitol-negative variant of Biovar 8, the majority (30/48) being Biovar 3. Of the 42 isolates for which somatic serotyping was conducted, 10% were Serovar 1, 79% were Serovar 3, 2% were Serovar 6,1, 2% were Serovar 12, and 7% could not be typed. All 48 isolates were confi rmed as P. multocida using a species-specifi c PCR. In the capsular multiplex PCR, 92% of isolates were Capsular (Cap) type A, 2% were Cap D, and 6% could not be typed. None of the samples were positive for the HSB or toxA genes. CONCLUSION: Serovars or capsular types of P. multocida associated with HS or PAR in pigs were not detected. Establishment of species-specifi c, capsular and toxin PCR assays allowed the rapid screening of isolates of P. multocida, while serotyping provided an additional tool for epidemiological and tracing purposes.

Reclassification of Pasteurella gallinarum, [Haemophilus] paragallinarum, Pasteurella avium and Pasteurella volantium as Avibacterium gallinarum gen. nov., comb. nov., Avibacterium paragallinarum comb. nov., Avibacterium avium comb. nov. and Avibacterium volantium comb. nov.

This paper describes a phenotypic and genotypic investigation of the taxonomy of [Haemophilus] paragallinarum, Pasteurella gallinarum, Pasteurella avium and Pasteurella volantium, a major subcluster within the avian 16S rRNA cluster 18 of the family Pasteurellaceae. An extended phenotypic characterization was performed of the type strain of [Haemophilus] paragallinarum, which is NAD-dependent, and eight NAD-independent strains of [Haemophilus] paragallinarum. Complete 16S rRNA gene sequences were obtained for one NAD-independent and four NAD-dependent [Haemophilus] paragallinarum strains. These five sequences along with existing 16S rRNA gene sequences for 11 other taxa within avian 16S rRNA cluster 18 as well as seven other taxa from the Pasteurellaceae were subjected to phylogenetic analysis. The analysis demonstrated that [Haemophilus] paragallinarum, Pasteurella gallinarum, Pasteurella avium and Pasteurella volantium formed a monophyletic group with a minimum of 96·8% sequence similarity. This group can also be separated by phenotypic testing from all other recognized and named taxa within the Pasteurellaceae. As both genotypic and phenotypic testing support the separate and distinct nature of this subcluster, the transfer is proposed of Pasteurella gallinarum, [Haemophilus] paragallinarum, Pasteurella avium and Pasteurella volantium to a new genus Avibacterium as Avibacterium gallinarum gen. nov., comb. nov., Avibacterium paragallinarum comb. nov., Avibacterium avium comb. nov. and Avibacterium volantium comb. nov. The type strains are NCTC 1118T (Avibacterium gallinarum), NCTC 11296T (Avibacterium paragallinarum), NCTC 11297T (Avibacterium avium) and NCTC 3438T (Avibacterium volantium). Key characteristics that separate these four species are catalase activity (absent only in Avibacterium paragallinarum) and production of acid from galactose (negative only in Avibacterium paragallinarum), maltose (negative only in Avibacterium avium) and mannitol (negative in Avibacterium gallinarum and Avibacterium avium).