Untreated Sheep Research Articles

Full-Length cDNA Cloning, Molecular Characterization and Differential Expression Analysis of Lysophospholipase I from Ovis aries.

Lysophospholipase I (LYPLA1) is an important protein with multiple functions. In this study, the full-length cDNA of the LYPLA1 gene from Ovis aries (OaLypla1) was cloned using primers and rapid amplification of cDNA ends (RACE) technology. The full-length OaLypla1 was 2457 bp with a 5′-untranslated region (UTR) of 24 bp, a 3′-UTR of 1740 bp with a poly (A) tail, and an open reading frame (ORF) of 693 bp encoding a protein of 230 amino acid residues with a predicted molecular weight of 24,625.78 Da. Phylogenetic analysis showed that the OaLypla1 protein shared a high amino acid identity with LYPLA1 of Bos taurus. The recombinant OaLypla1 protein was expressed and purified, and its phospholipase activity was identified. Monoclonal antibodies (mAb) against OaLypla1 that bound native OaLypla1 were generated. Real-time PCR analysis revealed that OaLypla1 was constitutively expressed in the liver, spleen, lung, kidney, and white blood cells of sheep, with the highest level in the kidney. Additionally, the mRNA levels of OaLypla1 in the buffy coats of sheep challenged with virulent or avirulent Brucella strains were down-regulated compared to untreated sheep. The results suggest that OaLypla1 may have an important physiological role in the host response to bacteria. The function of OaLypla1 in the host response to bacterial infection requires further study in the future.

Application of a body condition score index for targeted selective treatment in adult Merino sheep—A modelling study

This study aimed to establish whether sheep flock production losses due to nematode (worm) infections are typically greater in mature sheep selected for anthelmintic treatment at random compared to sheep selected for treatment based on low (poorer) body condition score (BCS). The study also examined the proportion of sheep in flocks that could be left untreated before production losses became evident, and projected worm egg pasture contamination. Sheep were monitored at two experimental sites in Western Australia (Mediterranean climate). Sheep were stratified for BCS, liveweight and faecal worm egg count (WEC) and allocated into treatment groups (treated or untreated), with equal numbers for each. Liveweight, BCS and WEC measurements were taken on 6 occasions at Farm A and 10 occasions at Farm B. Comparisons of sheep production (liveweight and BCS change) and pasture contamination potential (WEC) were conducted by generating “virtual flocks” of varying proportions sheep untreated (10%, 20%, 30%, 40%, and 50% untreated). For the comparison of the selection mode of sheep for treatment, the untreated sheep were either selected at random, or as the highest BCS animals at the commencement of observations. Univariate general linear models with least square difference post-hoc tests were used to examine differences between flocks for liveweight, BCS and WEC, and regression analysis was used to examine relationships between BCS and WEC, and liveweight and WEC. No difference in body weights was observed between flocks with varying proportions of ewes notionally left untreated at Farm B, and until more than 30% were left untreated at Farm A. There was no difference in BCS between flocks with varying proportions of ewes left untreated at either site. At no point were there differences in cumulative liveweight change or BCS between selection methods (BCS versus random) where the same proportion of sheep in virtual flocks were left untreated, suggesting that effort committed to individual BCS assessment would be of no benefit under these circumstances except for identifying low BCS sheep at risk of falling below critical limits associated with health or welfare risks. No consistent relationship between WEC and BCS or bodyweight was observed, indicating that BCS selection would have no lesser or greater impact on worm pasture contamination compared to random selection. Summer treatments based on a random selection index (with a minimum BCS limit), with up to 30% of adult sheep untreated can be expected to delay the development of anthelmintic resistance, with minimal adverse effect on sheep health or production.

Qualitative assessment of the entry of capripoxviruses into Great Britain from the European Union through importation of ruminant hides, skins and wool

Sheep pox and goat pox (SPGP) virus and lumpy skin disease (LSD) virus belong to the genus Capripoxvirus and cause disease with economic impacts in sheep/goats and cattle respectively. In 2013/14, outbreaks of SPGP were reported in sheep in Greece and Bulgaria and LSD outbreaks were reported in cattle in Turkey, Egypt and some countries in the Middle East. Clinical signs for both diseases include pox lesions, papules and scabs on the skin which may contain virus. This, together with the fact that Great Britain (GB) currently imports cattle hides, sheep skins and wool from European Union (EU) countries without the requirement for treatment prior to export, raises concern that capripoxviruses could be introduced into GB. A qualitative assessment presented here concluded that the current risk of entry of SPGP virus into GB through the importation of one untreated sheep skin, hide or wool bale from an EU Member State (MS) with similar flock prevalence to that in sheep in Greece in 2013/14 is low. In terms of SPGP virus levels, those infected sheep skins/hides entering GB are more likely to be from infected animals with normal skin (i.e., not showing lesions) and hence carrying lower levels of virus than those from animals showing papules and scabs which contain very high virus levels and are easier to detect. The predicted risk of importation of LSD virus per cattle hide/skin is also low (assuming LSD were to emerge in an EU MS with similar herd prevalence to that reported for SPGP in Greece in 2013/14). The levels of LSD virus on an infected cow's hide, if imported, may be very low. It is recommended that the risks for entry of capripoxviruses are recalculated if outbreaks occur elsewhere within the EU.

Effect of mesenchymal precursor cells on the systemic inflammatory response and endothelial dysfunction in an ovine model of collagen-induced arthritis.

Background and AimMesenchymal precursor cells (MPC) are reported to possess immunomodulatory properties that may prove beneficial in autoimmune and other inflammatory conditions. However, their mechanism of action is poorly understood. A collagen-induced arthritis model has been previously developed which demonstrates local joint inflammation and systemic inflammatory changes. These include not only increased levels of inflammatory markers, but also vascular endothelial cell dysfunction, characterised by reduced endothelium-dependent vasodilation. This study aimed to characterise the changes in systemic inflammatory markers and endothelial function following the intravenous administration of MPC, in the ovine model.MethodsArthritis was induced in sixteen adult sheep by administration of bovine type II collagen into the hock joint following initial sensitisation. After 24h, sheep were administered either 150 million allogeneic ovine MPCs intravenously, or saline only. Fibrinogen and serum amyloid-A were measured in plasma to assess systemic inflammation, along with pro-inflammatory and anti-inflammatory cytokines. Animals were necropsied two weeks following arthritis induction. Coronary and digital arterial segments were mounted in a Mulvaney-Halpern wire myograph. The relaxant response to endothelium-dependent and endothelium-independent vasodilators was used to assess endothelial dysfunction.Results and ConclusionArthritic sheep treated with MPC demonstrated a marked spike in plasma IL-10, 24h following MPC administration. They also showed significantly reduced plasma levels of the inflammatory markers, fibrinogen and serum amyloid A, and increased HDL. Coronary arteries from RA sheep treated with MPCs demonstrated a significantly greater maximal relaxation to bradykinin when compared to untreated RA sheep (253.6 ± 17.1% of pre-contracted tone vs. 182.3 ± 27.3% in controls), and digital arteries also demonstrated greater endothelium-dependent vasodilation. This study demonstrated that MPCs given intravenously are able to attenuate systemic inflammatory changes associated with a monoarthritis, including the development of endothelial dysfunction.

Abstract 9480: Persistent MR Reduction With Stabilization of LV Remodeling and Following Chronic Polymer-Mesh Device on Infarcted Myocardium in a Chronic Ischemic MR Model

Background: Mitral valve repair with ring annuloplasty for ischemic mitral regurgitation (IMR) is associated with significant recurrent IMR due to continued left ventricular (LV) remodeling as it does not directly address LV and mitral valve tethering. We aimed to assess chronic implantation of novel polymer based mesh device to reduce IMR reduction and reverse LV remodeling. Methods: Five Dorset sheep underwent ligation of second and third circumflex obtuse marginal branches to produce chronic IMR after 8 weeks. Poly-mesh was attached to the infarcted myocardium underlying the papillary muscle with a further 8 weeks follow-up. In addition, chronic IMR was produced in 5 untreated control sheep. IMR severity was assessed by 2D vena contracta width (VCW) and 3D vena contracta area (VCA). LV end-diastolic volume (EDV), end-systolic volume (ESV), ejection fraction (EF) by 2D echo and hemodynamic data were also obtained at each baseline, chronic MR, and chronic poly-mesh stages. Results: In all sheep, after poly-mesh attachment, IMR decreased and this reduction persisted over 8 weeks (VCW 0.42±0.09 to 0.08±0.12cm; VCA 0.13±0.08 to 0.05±0.08cmsq, p<0.01) with significant increase in Emax (1.1±0.5 to 2.9±0.7mmHg/ml, p<0.05). There were no significant changes in LVEF (45.4±6.7 vs 42.6±3.5%, p=NS). Importantly, LV EDV and ESV at chronic poly-mesh stage were reduced compared to the chronic MR stage (EDV 110±15 to 88±18ml, p<0.05; ESV 61±14 to 51±12ml, p=NS) and % increases of both EDV and ESV from chronic MR to chronic poly-mesh stage were significantly lower than those of untreated control sheep (%EDV change -22±11 vs 11±10%, p<0.01; %ESV change -10±14 vs 9±10%, p<0.05) (FIGURE). Conclusions: Chronic follow up after polymer-mesh device attachment shows the stabilization of mitral-LV complex in a chronic IMR model, causing persistent reduction of IMR and preventing continued LV remodeling.

Fasciola hepatica: A comparative survey of adult fluke resistance to triclabendazole, nitroxynil and closantel on selected upland and lowland sheep farms in Northern Ireland using faecal egg counting, coproantigen ELISA testing and fluke histology

In order to investigate the incidence and distribution of adult fluke resistance to the fasciolicide tricalbendazole (TCBZ) amongst populations of Fasciola hepatica in sheep flocks in Northern Ireland (NI), individual rectal faeces samples were collected from 3 groups of 20 sheep, before (pre-dose), and 21 days after (post-dose) treatment of the animals with TCBZ, nitroxynil or closantel, on each of 13 well-managed sheep farms distributed across the province. The efficacy of each flukicide was determined for each farm, using faecal egg count reduction (FECRT) and F. hepatica coproantigen ELISA testing. In certain flocks, 2 sheep with high pre-dose faecal egg counts (FEC) were killed 3 days and 21 days respectively after TCBZ treatment, and the histology of the fluke reproductive organs was compared with that of flukes from untreated sheep, and from sheep treated with nitroxynil or closantel 2 days prior to death, using haematoxylin and eosin (H&E) staining and an in situ hybridisation method (TdT-mediated dUDP nick end labelling [TUNEL]) to demonstrate apoptosis. Results from FECRT revealed that in all flocks with a high fluke burden, TCBZ was ineffective in treating chronic fasciolosis, and this finding was generally supported by the results of the coproantigen reduction test (CRT). The histology of reproductive organs of flukes from TCBZ-treated sheep in these flocks was normal, when compared with untreated flukes, and this, together with the FECRT and CRT findings, indicated a likely diagnosis of TCBZ resistance in all the flocks with a high fluke burden. In contrast, nitroxynil and closantel were found to be fully effective against TCBZ-resistant flukes in each of the flocks bearing a high chronic fluke burden. All of the flocks with a high fluke burden and TCBZ resistance were managed on lowland in the South and East of NI. Upland flocks, in the North and West, had low fluke burdens, or were clear of infection; and FECs were too low to allow valid resistance testing. The study highlights the high level of penetration of TCBZ resistance throughout F. hepatica populations in areas of intensively managed sheep production with a high level of fluke challenge. Further, it emphasises the importance of pre-emptive chemotherapeutic action against chronic fasciolosis, using flukicides effective against the egg-producing adult flukes to minimise pasture contamination for the next season's lamb crop. This study also exemplifies the use of several complementary methods (FECRT; CRT; fluke histology; comparative anthelmintic efficacy testing) for confirmation of a diagnosis of fluke drug resistance.

Effects of hygienic treatments during slaughtering on microbial dynamics and contamination of sheep meat

The aims of this study were to investigate bacterial dynamics in the sheep meat chain, from fleece to meat trimmings, using both quantitative and qualitative analyses, and to study the effects on microbial load associated with the hygienic interventions of: i) shearing sheep immediately before slaughter, ii) manual steam vacuum pasteurisation, iii) hot water pasteurisation of carcasses, followed by iv) chilling. A further aim was to provide evidence to determine whether or not unshorn sheep should be handled in a processing line separate from that of shorn sheep in Norwegian abattoirs. A total of 176 surface swab samples were collected from three sites along the value chain: i) on fleeces, ii) on carcasses at the end of the slaughter line, and iii) on carcasses after chilling for 24h, and 32 samples were collected from meat trimmings. The results showed that Aerobic Plate Counts (APC) were lower for the shorn group compared to the unshorn group, both on carcasses before chilling and after chilling (difference of 0.8 and 0.9logCFU/1000cm2 (p≤0.05), respectively) and in meat trimmings (difference of 0.5logCFU/g (p≤0.05)). Hygienic treatments were used on carcasses derived from unshorn sheep, and steam vacuum treatment reduced Escherichia coli, Enterobacteriaceae, and APC before chilling by 1.2, 1.0, and 0.6logCFU/1000cm2 (p≤0.05), respectively, and hot water pasteurisation, in addition to chilling, reduced E. coli, Enterobacteriaceae, and APC by 0.7, 1.0, and 0.9logCFU/1000cm2 (p≤0.05), respectively, compared with untreated carcasses. The effect of chilling was shown by the significant reduction of number of carcasses where E. coli were detected; from 65% (13/20) of the shorn group before chilling to 35% (7/20) after chilling, and from 90% (36/40) to 45% (9/20) of the unshorn group. Sequencing of the 16S rRNA gene derived from 316 colonies of Enterobacteriaceae showed a tendency for the relative proportion of the genus Escherichia/Shigella, compared with other genera within Enterobacteriaceae, to be greater for unshorn, untreated sheep than from the other groups at the sampling locations along the meat chain. The study showed that steam vacuum and hot water pasteurisation reduced the contamination of carcasses derived from unshorn sheep, down to the level of the shorn group, and thus can replace the separate processing line for unshorn sheep. Indeed, the low microbial contamination in meat trimmings for all groups indicates that the separate processing line is unnecessary.

Effective control of a suspected cyromazine-resistant strain of Lucilia cuprina using commercial spray-on formulations of cyromazine or dicyclanil.

To demonstrate the protection of Merino sheep from flystrike by Lucilia cuprina with cyromazine or dicyclanil in an implant study and in the field. In the implant study, sheep were treated with cyromazine or dicyclanil and implanted with 1st-stage larvae from a newly isolated field strain of L. cuprina (CYR-LS) or a reference strain (DZR50), then assessed over 3 days and compared with the implants on untreated control sheep. In the field study, weaner lambs were treated with cyromazine or dicyclanil and monitored weekly for flystrike over 18 weeks of grazing on the same farm from which the L. cuprina were isolated. Implant study: cyromazine (6%) provided effective protection against CYR-LS and DZR50 L. cuprina for a minimum of 13 and 10 weeks, respectively. Dicyclanil (5%) provided at least 18 weeks' protection against both strains. Field study: only 1 of 386 lambs in the cyromazine-treated group was struck in the first 14 weeks of the trial. No strikes occurred in the 198 sheep treated with dicyclanil (5%). Rainfall, temperature and flytrap data indicated consistent fly pressure during the study. Based on the results of these studies, there was no evidence of reduced susceptibility to cyromazine or dicyclanil and the periods of protection of sheep against L. cuprina were unaffected and consistent with the registered label claims.

Field trials assessing deltamethrin (Butox®) treatments of sheep against Culicoides species

Culicoides (Diptera: Ceratopogonidae) biting midges may transmit various diseases of economic importance, including bluetongue virus (BTV) and Schmallenberg (SV) virus, which affect ruminants. During the outbreak of BTV in central and northern Europe in 2006, and in the absence of BTV vaccines, many national veterinary services recommended the treatment of susceptible livestock with pyrethroids as a first-line defense against biting midges, although these insecticides were officially not registered and authorized for use against Culicoides midges. The efficacy of Butox® pour on (7.5 mg deltamethrin/mL) against biting midges was therefore evaluated in a double-blinded GCP field trial performed in Brandenburg, Germany. Forty female Merino sheep with an average body weight of 38 kg (±7 kg) were used for the study. Twenty randomly selected sheep were treated with 10 mL Butox® pour on. The remaining 20 sheep were left untreated and served as a control group. Midge collections took place in two separate drop traps covering two crush pens with three confined treated/untreated sheep standing inside, on weekdays at 1, 7, 14, 21, 28, and 35 days post treatment. A total of 19,057 midges were collected during this period. Midges were identified as belonging to the subgenus Avaritia, Fox (84.6%) and subgenus Culicoides, Latreille (15.4%). A total of 12,031 midges were collected inside the drop trap containing untreated sheep, in comparison to 7,026 midges collected from the vicinity of the treated sheep. Significantly, more midges had fed on control compared to treated sheep with 757 and 103 engorged midges, respectively. The results indicate that treatment of sheep with Butox® pour on provided a significant decrease in Culicoides feeding rates under field conditions for at least 35 days.

The variation of cancellous bones at lumbar vertebra, femoral neck, mandibular angle and rib in ovariectomized sheep

This study aimed to compare the variation of cancellous bones at four skeletal sites: lumbar vertebra, femoral neck, mandibular angle and rib in ovariectomized sheep. Sixteen adult sheep were randomly divided into two groups: eight sheep were ovariectomized served as experimental group; the other eight untreated sheep were served as control group. Bone mineral density was assessed by dual-energy X-ray absorptiometry on lumbar vertebrae at baseline and twelve months after ovariectomy. After 12 months, lumbar vertebrae L3 and L4, femoral necks, mandibular angles and the fourth ribs were harvested for micro-CT scanning, histological analysis and biomechanical test. The results showed that bone mineral density of lumbar vertebra decreased significantly in twelfth month (p<0.05). The results of micro-CT showed that the bone volume/total volume decreased by 45.6%, 36.1% 21.3% and 18.7% in lumbar vertebrae, femoral necks, mandibular angles and ribs in experimental group (p<0.05) respectively. The trabecular number showed the same downtrend (p<0.05). Histological analysis showed trabecular area/tissue area decreased by 32.1%, 23.2% and 20.7% in lumbar vertebrae, femoral necks and mandibular angles respectively (p<0.05), but no significant difference in ribs. Specimens elastic modulus from lumbar vertebra, femoral neck and mandibular angle were 952±76MPa (628±70MPa), 961±173MPa (610±72MPa) and 595±60MPa (444±31MPa) in control group (experimental group) respectively. These datum indicated that the sensibility of cancellous bones to oestrogen deficiency in ovariectomized sheep was site-specific on a pattern as follows: lumbar vertebra, femoral neck, mandibular angle and rib.

Carbon precursors of methane synthesis in the rumen of sheep dosed with ionophores

Rates of methane (CH4) production and the sources of carbon (C) for its synthesis were studied in four mature ewes when dosed with a CH4-mitigating ionophore ICI-111075, or monensin, or when untreated. The sheep were given 700 g/day of chaffed lucerne hay in equal portions every hour, before and during experiments in which 14C-labelled NaHCO3–, acetate, propionate, lactate and formate were infused intraruminally over 12 h and the specific radioactivity of C (SR) in each of these substrates was determined. During these infusions, the SR of material in the primary pool (the tracer infusion site) and in secondary metabolites of this material (secondary pools) approached asymptotic or ‘plateau’ values. The rate of infusion of 14CH4 (kBq/day) divided by the plateau SR value (kBq/g C) gave estimates of the rate of irreversible loss of CH4 (g C/day). These calculations indicated that CH4 production rate was reduced by 72% when sheep were dosed with ICI-111075 and by 58% when dosed with monensin. With monensin, the reduction in CH4 production was not associated with hydrogen (H2) accumulation in the rumen headspace gases whereas with ICI-111075, the decrease in CH4 production was associated with marked H2 accumulation in the headspace gases. When plateau SR were attained during the tracer infusions, the percentage ratio, (SR in any secondary pool: SR in the primary pool) gave an estimate of the fraction of C in that secondary pool derived from material of the primary pool. Calculated in this way, the percentage of CH4-C derived from rumen fluid carbon dioxide (CO2) averaged 59% in untreated sheep, and when sheep were dosed with ICI-111075, the corresponding percentage averaged 12%. These findings indicate there are sources of C for rumen CH4 synthesis other than rumen fluid CO2. However, there was no evidence that C from acetate, propionate, lactate or formate in rumen fluid were direct sources of the unidentified CH4-C. One plausible explanation for these findings is that CH4 is synthesised within naturally occurring microbial biofilms (attached to feed particles or the rumen wall) from CO2 that is produced locally by fermentation of unlabelled substrates within the biofilms. It is postulated that such pools of CO2 would be kinetically distinct and, during the infusion of 14C-labelled substrates, would exhibit a lower SR than the CO2 in the surrounding rumen fluid.

Prophylactic and therapeutic effects of garlic extract on Nerium oleander-induced arrhythmia: a new approach to antiarrhythmic therapy in an ovine model

Context. Oleander is a potent cardiotoxic plant and is a common cause of poisoning in human and animals. There is no affordable and cost-effective treatment for oleander poisoning. Objective. To evaluate the prophylactic and therapeutic effects of garlic extract (Allium sativum) on Nerium oleander (a potent cardiotoxic plant) intoxication in sheep. Materials and methods. Eight sheep were intravenously infused with an unsterilized hydro-ethanol extract of garlic (50 mg/kg) before or after receiving a lethal dose of dried leaves (as a powder) of oleander (100 mg/kg, orally). The cardiac rhythm was continuously monitored using biopotential wireless transmitters and telemetry system. For evaluation of therapeutic effects, six sheep received the lethal dose of oleander and were administered with garlic extract after development of cardiac arrhythmias. Subsequently, the survived animals from the therapeutic study (four sheep) were administered with oleander without receiving any medication. Some blood constituents, including total antioxidant capacity, malondialdehyde, and troponin I, were compared between treated and untreated animals. Results. Pretreatment with garlic extract reversed the arrhythmia caused by oleander to its previous normal rhythm in seven sheep, but, one sheep died of ventricular fibrillation. On therapeutic treatment, four sheep survived while two died of ventricular fibrillation. Dosing with oleander without receiving garlic extract resulted in death of all sheep due to ventricular fibrillation. Blood constituents did not show any significant changes between treated and untreated sheep, and before and after intoxication. Conclusions. Garlic extract reduced the case fatality from 100% to 12.5% and 33.3% as a prophylactic or therapeutic agent, respectively. Additionally, garlic extract delayed the time of onset of arrhythmias and prolonged the interval between intoxication and death of the animals. Garlic extract could be considered to be a potential and affordable antidote in oleander poisoning. However additional studies with a larger sample size and in other species need to be performed to confirm the results in this study.

Enzymatic Formulation Capable of Degrading Scrapie Prion under Mild Digestion Conditions

The prion agent is notoriously resistant to common proteases and conventional sterilisation procedures. The current methods known to destroy prion infectivity such as incineration, alkaline and thermal hydrolysis are harsh, destructive, environmentally polluting and potentially hazardous, thus limit their applications for decontamination of delicate medical and laboratory devices, remediation of prion contaminated environment and for processing animal by-products including specified risk materials and carcases. Therefore, an environmentally friendly, non-destructive enzymatic degradation approach is highly desirable. A feather-degrading Bacillus licheniformis N22 keratinase has been isolated which degraded scrapie prion to undetectable level of PrPSc signals as determined by Western Blot analysis. Prion infectivity was verified by ex vivo cell-based assay. An enzymatic formulation combining N22 keratinase and biosurfactant derived from Pseudomonas aeruginosa degraded PrPSc at 65°C in 10 min to undetectable level -. A time-course degradation analysis carried out at 50°C over 2 h revealed the progressive attenuation of PrPSc intensity. Test of residual infectivity by standard cell culture assay confirmed that the enzymatic formulation reduced PrPSc infectivity to undetectable levels as compared to cells challenged with untreated standard scrapie sheep prion (SSBP/1) (p-value = 0.008 at 95% confidence interval). This novel enzymatic formulation has significant potential application for prion decontamination in various environmentally friendly systems under mild treatment conditions.

Evaluation of Mosquito Responses to Pyrethroid Insecticides Topically Applied to Sheep

A rise in the incidence of mosquito-transmitted Cache Valley virus (CVV) in lambs in 2011 prompted a study to evaluate on-animal pyrethroid insecticides to reduce mosquito attacks on sheep. Using enclosure traps for 1 night per wk for 6 wk, we compared engorgement rates of mosquitoes given the opportunity to feed on untreated sheep and sheep treated with 1 Python insecticide ear tag (containing 10% zeta-cypermethrin and 20% piperonyl butoxide) per animal or 2 synergized permethrin body spray treatments (containing 2.5% permethrin and 2.5% piperonyl butoxide). During the 6-wk study, 18,920 mosquitoes were collected in the animal-baited enclosure traps. Thirteen species were identified from these collections with the floodwater species Aedes increpitus and Ae. idahoensis making up 68% of the total. Potential CVV vector species, making up 25% of the samples, included Ae. vexans, Ae. dorsalis, Culex tarsalis, and Culiseta inornata. Traps baited with untreated sheep collected 9,701 mosquitoes with 65% of these engorged. Traps baited with sheep treated with Python ear tags or permethrin spray collected 4,034 and 4,555, respectively, with engorgement rates of 23% and 35%. Blood feeding on ear-tagged sheep was significantly reduced by as much as 90% compared to the untreated sheep, and protection lasted 4 wk or longer. Permethrin spray treatments were most effective within 24 h after application and provided better protection against Ae. dorsalis than the Python tag. Effectiveness of the permethrin spray diminished 1 wk after the 2nd application was made. The effect of these treatments appeared to be repellency because negligible mosquito mortality was observed at the time of collection. Further evaluation of these insecticides under conditions of natural exposure to a mosquito-borne pathogen is warranted.

Thermal inactivation and variability of γ-glutamyltransferase and α-l-fucosidase enzymatic activity in sheep milk

We present a study of the thermal inactivation and variation of activity of two enzymes in sheep milk: γ-glutamyltransferase and α-l-fucosidase. We find that the thermal treatment of milk causes inactivation of α-l-fucosidase between 57 °C and 68 °C, and γ-glutamyltransferase between 68 °C and 80 °C. In untreated sheep milk, the highest level of enzymatic activity (EA) for these two enzymes is in the pH range 5.5–6.0 and 8.0–8.5, respectively. Skimming treatment causes the loss of 35% of the activity of γ-glutamyltransferase. No differences in the EA were ascribable to the diets evaluated in this study. We observed EAs of α-l-fucosidase and γ-glutamyltransferase over the entire lactation cycle, noting that the EA of the former increased from 81 to 207 U mL−1 (average 159 ± 30 U mL−1) while the EA of the latter from 3.12 to 4.89 U mL−1 (average 3.83 ± 0.34 U mL−1). Relationships between both enzymes and selected milk parameters (lipids, proteins, lactose, urea and somatic cells) are highlighted by Principal Component Analysis.

Developmental Programming: Postnatal Steroids Complete Prenatal Steroid Actions to Differentially Organize the GnRH Surge Mechanism and Reproductive Behavior in Female Sheep

In female sheep, estradiol (E2) stimulates the preovulatory GnRH/LH surge and receptive behavior, whereas progesterone blocks these effects. Prenatal exposure to testosterone disrupts both the positive feedback action of E2 and sexual behavior although the mechanisms remain unknown. The current study tested the hypothesis that both prenatal and postnatal steroids are required to organize the surge and sex differences in reproductive behavior. Our approach was to characterize the LH surge and mating behavior in prenatally untreated (Control) and testosterone-treated (T) female sheep subsequently exposed to one of three postnatal steroid manipulations: endogenous E2, excess E2 from a chronic implant, or no E2 due to neonatal ovariectomy (OVX). All females were then perfused at the time of the expected surge and brains processed for estrogen receptor and Fos immunoreactivity. None of the T females exposed postnatally to E2 exhibited an E2-induced LH surge, but a surge was produced in five of six T/OVX and all Control females. No surges were produced when progesterone was administered concomitantly with E2. All Control females were mounted by males, but significantly fewer T females were mounted by a male, including the T/OVX females that exhibited LH surges. The percentage of estrogen receptor neurons containing Fos was significantly influenced in a brain region-, developmental stage-, and steroid-specific fashion by testosterone and E2 treatments. These findings support the hypothesis that the feedback controls of the GnRH surge are sensitive to programming by prenatal and postnatal steroids in a precocial species.

A method for estimating the economic value of changes in the risk of breech strike

An economic model of the costs of sheep flystrike has been modified to examine the effect of a given change in the risk of breech strike as if it applied to the entire wool-producing industry, rather than to individual wool producers. Within each region (high rainfall, sheep/cereal and pastoral zones), the model sets the proportion of the sheep population in low, medium and high risk categories and calculates the estimated level of strike for untreated sheep in those groups. The costs related to breech strike can be calculated according to the original model. The risk of breech strike can then be adjusted to any required level, the proportion of sheep in each category adjusted and the costs recalculated according to the adjusted risk. The model estimated that a 50% reduction in the risk of breech strike would decrease the number of sheep requiring preventive treatment to about one-third of previous levels, but only provide a small reduction in costs related to struck sheep, due to the large increase in the number of sheep no longer receiving preventive treatment. There would be only small reductions in costs related to crutching and mulesing sheep unless much larger reductions in strike risk occurred. The overall benefit of a 50% reduction in the risk of strike would be $0.23–0.27 per sheep. If this reduction in strike risk occurred across all regions this could provide a benefit to the Australian sheep industry of $15 million. The model found that if mulesing was not used in any of the sheep then the use of preventive treatment would increase so that 93% rather than only 60% received chemical treatment, and reduction of the risk of breech strike by 50% provided benefits of $0.40 per sheep. Therefore, the estimated benefits would be greater than $0.27 per sheep in situations where the current level of risk is very high.

Dipping and jetting with tea tree (Melaleuca alternifolia) oil formulations control lice (Bovicola ovis) on sheep

The in vivo pediculicidal effectiveness of 1% and 2% formulations of tea tree (Melaleuca alternifolia) oil (TTO) against sheep chewing lice (Bovicola ovis) was tested in two pen studies. Immersion dipping of sheep shorn two weeks before treatment in both 1% and 2% formulations reduced lice to non detectable levels. No lice were found on any of the treated sheep despite careful inspection of at least 40 fleece partings per animal at 2, 6, 12 and 20 weeks after treatment. In the untreated sheep louse numbers increased from a mean (±SE) of 2.4 (±0.7) per 10cm fleece part at 2 weeks to 12.3 (±4.2) per part at 20 weeks. Treatment of sheep with 6 months wool by jetting (high pressure spraying into the fleece) reduced louse numbers by 94% in comparison to controls at two weeks after treatment with both 1% and 2% TTO formulations. At 6 and 12 weeks after treatment reductions were 94% and 91% respectively with the 1% formulation and 78% and 84% respectively with the 2% formulation. TTO treatment also appeared to reduce wool damage in infested sheep. Laboratory studies indicated that tea tree oil ‘stripped’ from solution with a progressive reduction in concentration as well as volume as more wool was dipped, indicating that reinforcement of active ingredient would be required to maintain effectiveness when large numbers of sheep are treated. The results of these studies suggest significant potential for the development of ovine lousicides incorporating TTO.

The comparative efficacy of abamectin, monepantel and an abamectin/derquantel combination against fourth-stage larvae of a macrocyclic lactone-resistant Teladorsagia spp. isolate infecting sheep

Anthelmintic resistance by gastrointestinal nematodes of sheep continues to be an issue of global interest. While the recent introduction in some countries of one or two new anthelmintic classes (amino-acetonitrile derivatives [AAD] and spiroindoles [SI]) has been welcomed, it is important that there is no relaxation in parasite control and the management of drug resistance. Monepantel (an AAD) was the first new anthelmintic to be approved for use (New Zealand, 2009) and was followed a year later in the same country by a combination of derquantel (a SI) and abamectin. The present study determined the efficacy of the new anthelmintic products and abamectin against fourth-stage larvae of macrocyclic lactone-resistant Teladorsagia spp. in lambs. Efficacies were calculated by comparing post-mortem nematode burdens of treated animals with those of untreated control sheep, and were 98.5, 86.3 and 34.0% for monepantel, abamectin/derquantel and abamectin, respectively. The nematode burdens of monepantel- and abamectin/derquantel-treated sheep were significantly lower than those sheep treated with abamectin and the untreated controls. Similarly, the burden of the monepantel group was significantly lower than that of the abamectin/derquantel group. These findings provide an opportunity to reinforce the recommendation that farmers and animal health advisors need to know the resistance status of nematode populations on subject farms to ensure effective control programs are designed and implemented. Such control programs should include an appropriate choice of anthelmintic(s), monitoring parasite burdens for correct timing of treatments, and pasture management to reduce larval challenge balanced with the maintenance of drug-susceptible populations in refugia.

Kidney-derived proteins in urine as biomarkers of induced acute kidney injury in sheep

Acute kidney injury (AKI) is a life-threatening condition for which an early diagnosis is problematic. The aim of the present study was to identify kidney-derived urinary proteins specific to AKI in sheep. AKI was induced in six sheep by an overdose of ketoprofen. Six untreated sheep served as controls. Urine samples were collected for up to 24 h after drug administration and pooled according to time and treatment. Tissue samples from kidney were taken immediately after euthanasia. Urinary proteins were separated by two-dimensional gel electrophoresis (2DE) and the proteins of interest were identified by mass spectrometry. Calbindin-D28k, retinol-binding protein 4 and CD1d were identified in ketoprofen-treated sheep, but not in controls. In addition, calbindin-D28k and CD1d were localized in kidney tissues by immunohistochemical staining. These preliminary results suggest that urinary calbindin-D28k and CD1d represent potential useful biomarkers of AKI, at least in sheep.