Academic literature on the topic 'Animal protection (incl. pests and pathogens)'

Productivity of crops grown for human consumption is at risk due to the incidence of pests, especially weeds, pathogens and animal pests. Crop losses due to these harmful organisms can be substantial and may be prevented, or reduced, by crop protection measures. An overview is given on different types of crop losses as well as on various methods of pest control developed during the last century.Estimates on potential and actual losses despite the current crop protection practices are given for wheat, rice, maize, potatoes, soybeans, and cotton for the period 2001–03 on a regional basis (19 regions) as well as for the global total. Among crops, the total global potential loss due to pests varied from about 50% in wheat to more than 80% in cotton production. The responses are estimated as losses of 26–29% for soybean, wheat and cotton, and 31, 37 and 40% for maize, rice and potatoes, respectively. Overall, weeds produced the highest potential loss (34%), with animal pests and pathogens being less important (losses of 18 and 16%). The efficacy of crop protection was higher in cash crops than in food crops. Weed control can be managed mechanically or chemically, therefore worldwide efficacy was considerably higher than for the control of animal pests or diseases, which rely heavily on synthetic chemicals. Regional differences in efficacy are outlined. Despite a clear increase in pesticide use, crop losses have not significantly decreased during the last 40 years. However, pesticide use has enabled farmers to modify production systems and to increase crop productivity without sustaining the higher losses likely to occur from an increased susceptibility to the damaging effect of pests.The concept of integrated pest/crop management includes a threshold concept for the application of pest control measures and reduction in the amount/frequency of pesticides applied to an economically and ecologically acceptable level. Often minor crop losses are economically acceptable; however, an increase in crop productivity without adequate crop protection does not make sense, because an increase in attainable yields is often associated with an increased vulnerability to damage inflicted by pests.

Farmers and growers, already under pressure to produce food under tighter constraints, face the omnipresent threat of new pests and diseases caused by new strains of fungal, bacterial or viral pathogens or their vectors, such as insects. The whitefly Bemisia tabaci is an example of such a threat in Europe (Bedford and Markham, 1993). It is feared that over-use of pesticides, fertilizers and growth regulators could create major environmental problems, but to survive without them will require other strategies, broadly referred to as ‘sustainable agriculture ’. Traditionally, durable disease resistance has required ingenious breeding programmes to introduce natural resistance genes from crop-related wild species into crops. However, we can now exploit the power, flexibility and specificity of recombinant DNA techniques (genetic manipulation) either to identify existing natural resistance genes and transfer them between unrelated crop species, or to create new and more effective resistance genes against one or more pathogens or pests. The rationale behind many of these ‘designer genes ’ has arisen from empirical field observations or from our deeper understanding of molecular and genetical aspects of the natural life-cycle of the pathogen.

"Soybean is an important economical crop used for human consumption, animal feed and industrial raw material. Also, soybean is succesfully used in crop roatations with the main cereal crops because it’s biological nitrogen-fixing capabilities. A great diversity of pests and diseases including nematodes, insects and phytopathogenic fungi are known to affect soybean crop. From plant emergence to grain maturity, pests Tetranychus urticae and Etiella zinckenella, pseudofungi Peronospora manshurica and fungi Fusarium sp. and Botrytis cinerea can cause economic damage. In this study was evaluated the influence of tillage systems, different types of fertilizers and pesticide treatments on the most economical important diseases and pests of soybean crop in a field experiment at Agricultural Research and Development Station (ARDS Turda) in the climatic conditions of 2020. Teo TD, an early maturing soybean variety created at ARDS Turda, was used for the experiment. Based on the assessments made, soil tillage system, fertilization and control of pests and diseases have differently influenced the downy mildew and T. urticae attack. To reduce the attack of pathogens in soybean crop, the best technological option is plowing, balanced fertilization and application of fungicides, either without or in combination with an insecticide. The T. urticae populations developed on mineral, organic and green fertilized plants. Integrated control methods must combine agro-technical measures with the application of chemical treatments in accordance with the warning of the appearance of the first adults. "

We review interactions between crop protection practices (developed to control plant pathogens and invertebrate pests) and human fungal infectious diseases. Unlike viral, bacterial and parasitic infections, fungal infections in humans are usually only superficial in healthy individuals, but can become invasive and pose serious risks to immunosuppressed individuals. Although their global impact is less than that of other infectious diseases, human fungal infections still pose serious public health issues. For instance, the use of synthetic agricultural fungicides, particularly the azole class, under conventional intensive, or efficiency improvement-based crop protection practices, is at risk as far as antimicrobial resistance is concerned, due to cases of cross-resistance to clinical azoles used to treat pulmonary aspergillosis, candidiasis and cryptococcocis. In this respect, the One Health approach, originally designed for other types of human pathogens, looks relevant for human pathogenic fungi. Additionally, some entomopathogenic fungi used as biocontrol products against crop pests in a substitution-based approach, may be potentially pathogenic to humans. Very few examples of redesign-based practices (i.e. Agroecological Crop Protection) emerged from our analysis on human fungal diseases. However, discontinuing agricultural azole fungicides (as practiced on organic farms, and which may to some extent be related to the redesign strategy) appears to be the best way to reduce selection pressure and hence the level of azole-resistant human pathogenic fungal strains in the environment.

Insects and ectoparasites are causes for major concern throughout the world due to their economic and welfare impacts on livestock agriculture. Current control measures involve chemicals such as acaricides which pose challenges like chemical resistance and longer withholding periods. To enable more sustainable agriculture practices, it is important to develop technologies that combine targeted effectiveness with minimal environmental footprint. RNA interference (RNAi) is a eukaryotic process in which transcript expression is reduced in a sequence-specific manner. This makes it a perfect tool for developing efficient and effective biological control against pests and pathogens. Double-stranded RNA (dsRNA) is the key trigger molecule for inducing RNAi; this concept is widely studied for development of RNA-based biopesticides as an alternative to chemical controls in crop protection for targeting pests and pathogens with accuracy and specificity. In this review, we discuss key advances made using RNAi technology and how they can be applied to improve health in livestock industries. This includes research focused on different delivery mechanisms of dsRNA, important developments in regulatory frameworks, and risk identification, that will enable the future adoption of RNAi technologies to improve animal health.

Asexual Epichloë spp. fungal endophytes have been extensively studied for their functional secondary metabolite production. Historically, research mostly focused on understanding toxicity of endophyte-derived compounds on grazing livestock. However, endophyte-derived compounds also provide protection against invertebrate pests, disease, and other environmental stresses, which is important for ensuring yield and persistence of pastures. A preliminary screen of 30 strains using an in vitro dual culture bioassay identified 18 endophyte strains with antifungal activity. The novel strains NEA12, NEA21, and NEA23 were selected for further investigation as they are also known to produce alkaloids associated with protection against insect pests. Antifungal activity of selected endophyte strains was confirmed against three grass pathogens, Ceratobasidium sp., Dreschlera sp., and Fusarium sp., using independent isolates in an in vitro bioassay. NEA21 and NEA23 showed potent activity against Ceratobasidium sp. and NEA12 showed moderate inhibition against all three pathogens. Crude extracts from liquid cultures of NEA12 and NEA23 also inhibited growth of the phytopathogens Ceratobasidium sp. and Fusarium sp. and provided evidence that the compounds of interest are stable, constitutively expressed, and secreted. Comparative analysis of the in vitro and in planta metabolome of NEA12 and NEA23 using LCMS profile data revealed individual metabolites unique to each strain that are present in vitro and in planta. These compounds are the best candidates for the differential bioactivity observed for each strain. Novel endophyte strains show promise for endophyte-mediated control of phytopathogens impacting Lolium spp. pasture production and animal welfare.

The tomato is one of the most nutritious, economically important, and delicate vegetables grown in the world. It is highly susceptible to insect pests and microbial pathogens. The tomato leafminer moth, Tuta absoluta Meyrick, is the current impediment to tomato production in the world. The insect showed invasive and notorious behavior and was affecting tomato production. To control this insect, the application of synthetic insecticides is seen as the primary solution. However, during the feeding stage, larvae hide within mined leaf mesophyll and bored fruits from chemical spray, besides fast developing resistance to several insecticides. Such characteristics of the insect reduced the effectiveness of the chemical control efforts. Currently, the natural, or ecofriendly pest control method is gaining the momentum to minimize the application of synthetic insecticide against this devastating insect. Studies showed that botanical extracts (phytochemicals) and natural enemies such as parasitoids, predators, entomopathogenic nematodes, entomopathogenic fungi, and entomopathogenic bacteria are effective for controlling T. absoluta. As a result, the basic attributes of the above-mentioned natural agents and their potential to control T. absoluta have been briefly discussed in this review. However, due to disease (pests), the expected outcome for the subsectors is still low. Therefore, the pinpointing of major diseases and pests and their control measures would help to significantly improve the crop production technology used by smallholder farmers and thereby sustainably improve tomato production in Ethiopia.

The introduction of intensive technologies in modern agricultural production involves the widespread use of chemicals to protect plants and animals, including insecticides and acaricides. However, the development of arthropods resistance to the used drugs significantly reduces the efficiency of the chemical method of controlling insects and mites. The article describes the basic mechanisms of the development of resistance on which the study of a comprehensive strategy of combating pests and pathogens in plants and animals based on. Multiple resistance in case of revealed resistance of target objects to multiple substances is of particular importance. Necessity for the development of new insecticidal and acaricidal preparations as well as the improvement of the principles of their application to slow down the development of resistance in arthropods is required. In animal husbandry, the problem is compounded by developing parallel resistance of infectious agents to antimicrobial agents. This requires the development of a scientific-based methodology of pharmacological prevention and treatment of infectious and parasitic diseases of animals, as well as chemical protection of plants against pests and diseases.

AbstractBiological control is defined broadly as the “use of natural or modified organisms, genes, or gene products” to reduce the effects of pests and diseases. Physical control is the use of tillage, open-field burning, heat-treatment (pasteurization), and other physical methods, usually to eliminate pests or separate them from the crop. Chemical control is the use of synthetic chemical pesticides to eliminate pests or reduce their effects. The many approaches to biological control can be categorized conceptionally into 1) regulation of the pest population (the classical approach), 2) exclusionary systems of protection (a living barrier of microorganisms on the plant or animal that deters infection or pest attack), and 3) systems of self-defense (resistance and immunization). The agents of biological control include the pest- or disease-agent itself (sterile males or avirulent strains of pathogens), antagonists or natural enemies, or the plant or animal managed or manipulated (immunized) to defend itself. The methods range from 1) conserving and making maximum use of indigenous (resident) biological control through cultural practices, 2) making one-time or occasional introductions of genes or natural enemies that are more or less self-sustaining and 3) making repeated introductions of a biocontrol agent (e.g. a microbial pesticide). Biological, physical, and chemical treatments and pest controls can be integrated into holistic plant-health care also known as integrated crop and pest management. Eight principles of plant health care are offered: 1) know the production limits of the agroecosystem; 2) rotate the crops; 3) maintain soil organic matter; 4) use clean planting material; 5) plant well-adapted, pest-resistant cultivars; 6) minimize environmental and nutritional stresses; 7) maximize the effects of beneficial organisms; and 8) protect with pesticides as necessary.

With a projected population of 10 billion by 2050, an immediate priority for agriculture is to achieve increased crop yields in a sustainable and cost-effective way. The concept of using a transgenic approach was realized in the mid-1990s with the commercial introduction of genetically modified (GM) crops. By 2010, the global value of the seed alone was US $11.2 billion, with commercial biotech maize, soya bean grain and cotton valued at approximately US $150 billion. In recent years, it has become evident that insect-resistant crops expressing δ -endotoxin genes from Bacillus thuringiensis have made a significant beneficial impact on global agriculture, not least in terms of pest reduction and improved quality. However, because of the potential for pest populations to evolve resistance, and owing to lack of effective control of homopteran pests, alternative strategies are being developed. Some of these are based on Bacillus spp. or other insect pathogens, while others are based on the use of plant- and animal-derived genes. However, if such approaches are to play a useful role in crop protection, it is desirable that they do not have a negative impact on beneficial organisms at higher trophic levels thus affecting the functioning of the agro-ecosystem. This widely held concern over the ecological impacts of GM crops has led to the extensive examination of the potential effects of a range of transgene proteins on non-target and beneficial insects. The findings to date with respect to both commercial and experimental GM crops expressing anti-insect genes are discussed here, with particular emphasis on insect predators and parasitoids.

A reduction in voluntary feed intake is a major factor in the lost productivity of grazing lambs infected by gastrointestinal parasites yet the mechanisms involved are poorly understood. Potential pathways involved in parasite-induced feed intake depression were investigated in lambs with minimal previous exposure to parasites and artificially infected by the small intestinal parasite Trichostrongylus colubriformis. Six in vivo experiments were conducted on lambs housed in individual pens or metabolism crates with similar feeding and experimental procedures. In Experiment 1 (Chapter 4) the effect of T. colubriformis infection on short term feed intake in lambs and of some pharmacological agents on feed intake depression were investigated. Prior to and for the duration of infection, lambs were fed once per day and feed intake recorded at regular intervals over the day (8 h). Following the onset of feed intake depression in the infected group (9 weeks after commencing dosing), all animals were treated with an analgesic (codeine phosphate per os), an anti-inflammatory agent (indomethacin per os), a CCK antagonist (L364-718 by subcutaneous injection) or saline (control) in a replicated Latin square design (n = 8). Although the pattern of feed consumption was similar in infected and non-infected lambs, average daily intake was reduced 32 % and short term intake (recorded at 10 minute intervals for the first hour of feeding, 15 minute intervals for the second hour and hourly for the next 6 hours of feeding) reduced 40 % by infection. This identified the key component by which intake was depressed and enabled the use of a short term intake model and short duration of action compounds to identify the pathways involved in intake depression in this sequence of experiments. None of the pharmacological treatments increased intake in the infected group. These results suggest a reduction in the rate of consumption due to reduced hunger signals, rather than change of meal eating patterns, is the major cause of feed intake depression. Specific conclusions about the pathways investigated using the pharmacological agents could not be obtained. Experiment 2 (Chapter 5) was designed to investigate the roles of pain and osmolality on feed intake depression. Digesta samples collected prior to and during parasite infection and before and after feeding had similar osmolalities (240-260 mosmol/l) which indicated that feeding or infection had no effect on osmolality of digesta. Following the onset of feed intake depression in infected animals, all animals were treated in a Latin square design (n = 4) with no treatment, saline, local anaesthetic (xylocaine) or analgesic (codeine phosphate) solution 15 minutes before feeding, by slow injection into the duodenum. There was no effect of these treatments on food intake. In the second part of the experiment, hyperosmotic solutions (mannitol and NaCI) markedly depressed short term intake in non-infected animals, suggesting a role for osmoreceptors in intake regulation. However these effects were not blocked by local anaesthetic so the depressed intake may have resulted from generalised malaise rather than from specific osmoreceptor effects. In Experiment 3 (Chapter 6) the role of peripheral CCK on intake depression was examined by a dose-response study utilising the CCK antagonist, loxiglumide. Intravenous injection of 5, 10 or 20 mg/kg LW of loxiglumide to infected lambs 10-15 minutes before feeding (n = 6) had no effect on feed intake at any of the dose levels. In experiment 4 (Chapter 7) loxiglumide was infused intravenously for 10 minutes (30 mg/kg/h) before feeding and for the first 2 h (10 mg/kg/h) after feed was offered to minimise any effect of the rate of clearance of loxiglumide on the lack of feed intake response. As well, the rate of marker disappearance from the abomasum was recorded in both infected and non-infected animals. Continuous infusion of loxiglumide did not attenuate parasite induced intake depression nor did it have any effect on abomasal emptying. Abomasal volume was reduced by infection (66.3 vs 162 ml) as was the fractional outflow rate (2.2 vs 2.8 ml/min) but these differences were accounted for by the lower level of feed intake in the infected animals. In Experiment 5 (Chapter 8) brotizolam, a benzodiazepine appetite stimulant, thought to act on the hypothalamus, was administered in a dose-response study to infected and non-infected animals (n = 4) immediately prior to feeding or following termination of the first meal (45 minutes after feeding) and the feed intake response recorded. Brotizolam elevated both the short term (0-0.75 h), daily (22 h) intake and all time intervals in the first 5 h after feeding in infected and non-infected animals when administered after the first meal but when administered prior to feeding elevated intake only over the first 6 h of feeding. In both cases the magnitude of the response was greater in infected animals than in non-infected animals. Brotizolam appeared to increase the rate of eating without having a major impact on meal eating patterns when administered before feeding. Where administration was after the first meal, the effect was due to an "extra" meal being consumed. These findings showed that infected animals can respond to central stimulators of intake although the mechanism of the response is not known. Opioids were implicated in intake depression as the rate of intake rather than meal patterns appeared to be the major parameter depressed under parasitism. This was examined in experiment 6 (Chapter 9) where animals (n = 6) were fasted for 26 h or not fasted, then treated with saline (control), brotizolam (intake stimulant) or naloxone (opioid antagonist) immediately prior to feeding. Fasting stimulated feed intake in the short term (100 % increase in 75 min) and over the day (12 % increase) in both infected and non-infected animals. Following fasting, infected animals ate a similar amount of feed to the non-infected, fasted animals and more than the non-infected, non-fasted animals. The signals resulting from a one day fast were sufficient in the short term to override parasite induced mechanisms causing feed intake depression. Naloxone suppressed the intake stimulatory effects of a 26 h fast in both infected and non-infected animals, which supports a role for endogenous opioids as hunger signals. Where animals were not fasted, naloxone reduced intake only in the non-infected animals which suggested endogenous opioid levels may be lower in infected animals than in non-infected animals. In the final experiment (Experiment 7, Chapter 10) the role of central hunger and satiety mechanism were investigated. Infected and non-infected animals (n = 6) were treated with naloxone or saline by intravenous injection, or saline and met-enkephalinamide (an opioid analogue) by intracerebral infusion, or naloxone and the opioid analogue simultaneously to investigate the role of central opioids in feed intake depression. To determine the role of CCK induced satiety signals on feed intake at a central level, loxiglumide and CCK were infused separately and in combination for 30 minutes prior to feeding and for the first 60 minutes of feed on offer, into a lateral cerebral ventricle of the brain of infected and control animals (n = 6). The opioid analogue tended to increase intake in infected animals but the effect was not significant probably because the dose used was too low to elicit a response in sheep. Naloxone depressed intake only in the infected animals, which conflicted with the results of Experiment 4. As a consequence these results were inconclusive because of the single low dose of opioid analogue used and the conflicting naloxone responses. CCK alone depressed intake by 39-52 % only in infected animals and this effect of the 90 minute infusion was evident over the 8 h short term recording period. Loxiglumide attenuated the feed intake depressive effects of CCK in the infected animals to the extent that intake was elevated above control levels. Loxiglumide alone was an intake stimulant in both infected and non-infected animals. Intake was increased over the entire 8 h but mostly in the second hour when intake was increased by 188 % in infected animals and by 16 % in the non-infected animals and resulted in almost continuous eating. These results showed loxiglumide will temporarily block the effect of parasite infection on feed intake in sheep when administered centrally and the fact that it blocked the effects of exogenous CCK on intake indicated that the effect is mediated via CCK receptors. In conclusion GIT parasite infection reduced both short term and daily feed intake apparently by a change in rate of intake rather than any alteration in meal patterns. It was further suggested that anyone of a number of potential peripheral pathways, including changes to osmolality, gut emptying, pain and inflammation of the gut, alone is not involved in anorexia in sofar as the compounds used could block these factors and the results support the idea that intake depression is mediated via a central mechanism. Intake in infected animals responded to a much greater extent when fasting, i.c.v. loxiglumide or brotizolam were employed. Feed intake thus appears to be regulated through the same mechanisms in infected and non-infected animals. The results from compounds affecting the central mechanism suggest central CCK receptors are important in parasite induced anorexia, possibly by changing the onset of satiety or by interacting with endogenous opioids to reduce the rate of feed intake. Secondly reduced endogenous opioids may be causing the reduction in the rate of feed consumption alone or as a result of other interactions. It was concluded that intake in parasitised animals could be increased to that of control animals by employing procedures and compounds thought to act on the hypothalamus.

Footrot is a contagious hoof disease of ruminants. It is endemic in New Zealand and throughout sheep and goat farming regions of the world. The disease results from a mixed bacterial infection, but the essential agent is Dichelobacter nodosus, a Gram-negative, anaerobic bacterium that possesses type-IV fimbriae on its surface. Genetic variation in the fimbriae of D. nodosus was investigated in this study. Using the polymerase chain reaction (PCR), the variable region of the gene encoding the fimbrial subunit (fimA) was amplified from bacterial DNA extracted from footrot lesions. Different fimA amplimers were differentiated by single-strand conformation polymorphism (SSCP) analysis. In conjunction with DNA sequencing, 15 unique sequences of D. nodosus fimA were obtained from 14 footrot samples taken from 6 farming regions throughout New Zealand. When these sequences were compared to fimA of known serogroups, it revealed that there were at least 15 D. nodosus strains, representing 8 serogroups, present on New Zealand farms. The predominant serogroup was B which contained 6 strains, followed by serogroups F, H and G. No strains from serogroups D and I were detected in this investigation. Twelve out of the 15 New Zealand D. nodosus strains had fimbriae different to those previously reported and the presence of multiple strains on a single hoof was common (86% of samples). The fimA sequences from the 12 D. nodosus strains incorporated into the footrot vaccine currently available in New Zealand were determined. A primer set targeting the relatively conserved fimA regions and based on the published sequence of serogroup M Nepalese isolates (designated M-Nep), failed to amplify fimA from the vaccine serotype M strain (designated as M-SPAHL). When the downstream primer was substituted with a primer that was specific for other serogroups of D. nodosus, the fimA gene was successfully amplified. Cloning followed by DNA sequencing, revealed that M-SPAHL fimA was different to M-Nep fimA. The predicted amino acid sequence of M-SPAHL fimA did not show homology to any known serogroups or serotypes. The most similar sequence was from serotype F1, and not M-Nep. The sequence difference between M-SPAHL and M-Nep was larger than that expected within a serogroup. The consequences of serological relatedness and sequence dissimilarity are discussed. Only eight of the 15 New Zealand field strains had fimbriae identical to those of the vaccine strains, while the remaining seven strains possessed different fimbriae. In addition, the vaccine contained two more D. nodosus strains, representing two sera groups, that were not found on the New Zealand farms investigated in this study. This may, to some extent, explain why the current footrot vaccine is at times less efficient in New Zealand. Another 17 footrot samples were screened for new or additional D. nodosus strains. Two PCR amplimers (designated X and Y) derived from footrot samples generated SSCP patterns different to those of previously identified strains. DNA sequencing revealed that these two fragments possessed novel sequences. The upstream of X (nt 1-183) was identical to serotype M1 while its downstream (nt 223-414) was identical to serotype F1; the upstream of Y (nt 1-116) was identical to serotype E1 whereas its downstream (nt 148-423) was identical to serotype F1. A 14-mer sequence consisting of two partially overlapping Chi-like sequences, 5'-GCTGGTGCTGGTGA-3', was also found in these fragments. Two primer sets with the downstream primer specific for serotype Fl and the upstream primer specific for serotype M1 or E1, produced PCR products of the expected sizes from the footrot samples from which fragments X and Y were isolated, respectively. These primer sets did not appear to amplify artificially mixed genomic DNA from serotypes M1 and F1 or E1 and F1. However, when the reactions were re-amplified, PCR recombination artefacts were observed, suggesting that PCR recombination does occur, but at a low frequency. It therefore seems more likely that fragments X and Y reflect genuine fimA sequences of D. nodosus which have resulted from in vivo DNA recombination, than from a PCR recombination artifact. The genetic capability for recombination at the fimbrial subunit locus may therefore endow D. nodosus with the ability to alter its antigenic appearance. D. nodosus strains present in footrot lesions can be genotyped using a PCR-SSCP/sequencing technique. However, this typing technique requires cloning and screening of D. nodosus fimA sequences, which is both laborious and costly. A rapid molecular typing system for D. nodosus was therefore developed in this study. A close examination of available D. nodosus fimA sequences revealed regions that appear to be specific for serogroups and serotypes. These regions were used to design a panel of sequence-specific oligonucleotide probes (SSOPs), and a rapid and accurate D. nodosus typing system using PCR and reverse dot-blot hybridisation (PCR/oligotyping) was subsequently developed. The variable region of D. nodosus fimA, amplified and labelled with digoxigenin (DIG) in a single multiplex PCR amplification, was hybridised to a panel of group- and type-specific, poly-dT tailed oligonucleotides that were immobilised on a nylon membrane strip. A mixture of positive control poly-dT tailed oligonucleotides was also included on the membrane. After hybridisation the membrane was washed to a defined specificity, and DIG-labelled fragments that had hybridised were detected. The specificity of the oligonucleotides was verified by the lack of cross-reactivity with D. nodosus fimA sequences that had a single base difference. DNA from 14 footrot samples previously genotyped by PCR-SSCP/sequencing, was assayed using the PCR/oligotyping technique. All types of D. nodosus which had been detected previously with a PCR-SSCP/sequencing method were detected by this procedure. However, for three of the 14 footrot samples, PCR/oligotyping detected additional types of D. nodosus. Further PCR amplification using type-specific primers, confirmed that these types were present in the original footrot samples. These results indicate that PCR/oligotyping is a specific, accurate, and useful tool for typing footrot samples. In combination with a rapid DNA extraction protocol, D. nodosus present in a footrot sample can be accurately genotyped in less than two days. Individual animals from the same farm, or the same paddock, were often infected by different strains of D. nodosus. This suggests a host role in mediating footrot infection, or that the interaction between the pathogen and the host is important. In order to better understand the interaction between the bacterium and the host, two polymorphic ovine class II MHC genes DQA1 and DQA2, which have been previously shown to be important in footrot infection, were also investigated in this study. PCR-SSCP/sequencing analysis of the DQA1 locus revealed ten unique ovine DQA1 sequences, with five of them being newly identified. This increases the number of known ovine DQA1 alleles from 8 to 13 (including a null allele), implying a high level of polymorphism at the ovine DQA1 locus. D. nodosus present on 20 footrot infected sheep from the same flock were genotyped, together with the ovine DQA1 and DQA2 genotypes of their hosts. Preliminary results showed that sheep with the same DQA1 and DQA2 genotypes tended to be infected by similar types of D. nodosus. Different types of D. nodosus were generally found on sheep with different genotypes at either the DQA1 or the DQA2 locus. This suggests the diversity in D. nodosus infection may be associated with the heterogeneity in the host MHC. However, as only a small number of animals from the same sire were analysed, further investigation is needed to gain a better understanding of the interaction between D. nodosus and the host MHC.

Aims to determine whether the toxins of Pimelea can be immunogenic and if so, to investigate whether antibodies raised to the toxins offered any protection against Pimelea toxicity. The study also provides opportunity to examine the in vitro contractile effects of the toxins on ovine pulmonary venule preparations.. Poisoning by Pimelea plants causes annual production losses exceeding $10 million to the Queensland cattle industry. Poisoning is caused either by ingestion of plants or by inhalation of dried plant debris. Daphnane and tigliane diterpenoid toxins found in the various Pimelea species are absorbed into the blood and cause pathological responses in target tissues through prolonged activation of protein kinase C isoenzymes. Simplexin, huratoxin and derivatives are the most toxic compounds prevalent in Pimelea species causing poisoning of cattle. Cattle are more vulnerable than sheep and horses to the toxins because the bovine pulmonary venous system has distinct sphincter -like arrangements in the smooth muscle coating of the venules. Prolonged contraction of the pulmonary system leads to oedema, jugular distension and right failure. At the present time there is no effective means of controlling pimelea poisoning in cattle, apart from removing animals from pastures infected with the plant. The main objectives of the research program described this thesis was to determine whether the toxins could be made immunogenic and if so, investigate whether antibodies raised to the toxins offered any protection against Pimelea toxicity. The study also provided the opportunity to systematically examine the in vitro contractile effects of the toxins on bovine pulmonary venule preparations. These organ bath experiments also allowed investigation of the efficacy of purified anti-toxin antibodies and potential antagonists of the toxins. The family of Pimelea diterpene toxins were isolated from dried Pimelea trichostachya plant material through a combination of solvent extraction and partition, column chromatography and preparative reverse phase HPLC. The toxins were conjugated to carrier proteins (HSA or ovalbumin) through a succinylate linker. Two structurally similar and commercially available daphnane orthoesters, mezerein and resiniferinol were also conjugated to carrier protein. Two groups of rabbits were vaccinated with the Pimelea- derived protein conjugates. ELISA methodology was developed to detect the presence of specific antibodies against Pimelea toxins from the serum of the vaccinated animals. After confirming the presence of specific IgG against Pimelea toxins in vaccinated rabbits, three groups of cattle were vaccinated with vaccines prepared from Pimelea, mezerein and resiniferinol protein conjugates. Cattle were vaccinated with the mezerein and resiniferinol conjugates to investigate the possibility of using commercially available compounds for vaccine preparation. The ELISA results for the vaccinated cattle showed the presence of specific antibodies which recognised toxin-protein conjugates coated to the plates. The ELISA cross reactivity data also showed that antibodies raised in animals vaccinated with mezerein and resiniferinol conjugates also recognised the Pimelea toxins. Vaccinated and control cattle were exposed to Pimelea trichostachya plant material to establish whether the antibodies had any protective effects. Both groups of cattle developed symptoms of Pimelea poisoning over two weeks, although these animals were dosed with much higher Pimelea concentrations than would be expected under normal grazing conditions. Various blood parameters were monitored throughout this experiment and there were some trends evident under the lower dose regimes that vaccinated animals were less affected than controls.The organ bath studies of isolated bovine pulmonary venule preparations showed that the EC50 toxin concentration required to elicit half maximal contraction (relative to the 1.0 M 5-HT response) was in the sub-micromolar range. Purified rabbit anti-toxin IgG was shown to attenuate the contractibility of bovine pulmonary venules in response to Pimelea toxins (EC5O) a dose dependent manner, whereas the purified bovine anti-toxin IgG lacked any efficacy. Nux vomica CM tincture (at doses above 200 L in the 25Ml baths) completely inhibited contraction response to toxins (EC50).

Bovine respiratory disease (BRD) is an ongoing health and economic issue in the dairy and beef cattle industry. Also, there are multiple risk factors that make an animal susceptible to BRD and it's diagnosis and treatment is a challenge for producers. Four bacterial species, Mannheimia haemolytica, Pasteurella multocida, Histophilus somni, and Mycoplasma bovis have been associated with BRD mortalities. Hence, this study aims to characterize the cattle nasal microbiome as a potential additional diagnostic method to identify animals suspected to have a lung infection. Quantitative PCR and 16S rRNA gene sequencing were used to determine the bacterial load of these four bacterial pathogens in the nasal microbiome of apparently healthy (N=75) and (N=58) affected by BRD Holstein steers. We then sought to identify a value or equation that could be used to discriminate between BRD and healthy animals using a Linear Discriminant Model (LDA). Additionally, co-occurrence between commensal bacterial and BRD-pathogens were also identified. Cattle diagnosed with BRD presented lower richness, evenness and phylogenetic diversity than healthy pen-mates. Bacterial species and genera Truperella pyrogenes and Bibersteina were increased in the BRD group, and the species Mycoplasma bovirhinis and Clostridium sensu stricto increased in the healthy group. Prevalence of H. somni (98%) and P. multocida (97%) were the highest regardless of disease diagnosis in all the samples. Prevalence of M. haemolytica (81 vs. 61%) and M. bovis (74 vs. 50.7%) were higher in the BRD group. The bacterial density of M. haemolytica and M. bovis was also higher in the BRD group, whereas Histophilus somni was lower in the BRD group. Five different models were tested using LDA, and one model produced a sensitivity and specificity of 60% and 81% agreement with diagnosis based on animal symptoms. Co-occurrence analysis demonstrated that the nasal microbiome members are more likely to interact with each other than associations between BRD-pathogens and nasal microbiome members. This study offers insight into the BRD-pathogens prevalence and difference in nasal microbiome between healthy and BRD animals and provides a potential platform for future studies and potential pen-side diagnostic testing.