THE CHANGES IN PROTEIN CONCENTRATION AND BACTERIA OF FLEECE AND SKIN DURING THE DEVELOPMENT OF FLEECE-ROT AND BODY STRIKE IN SHEEP

The sequence of events in the development of fleece-rot and body strike in sheep is described. When sheep were wetted by rain, vast increases in the numbers on microorganisms on the fleece and skin surface occured. Frequently, only one bacterial genus, Pseudomonas, proliferated and produced a bacteriostat to inhibit the growth of other resident flora. The development of fleece-rot lesions was characterised by a sudden substantial increase od soluble protein, presumably plasma protein, onto the skin surface. Pseudomonas spp proliferated on the skin surface and frequently produced extracellular, green pigment. First instar blowfly larvae were observed subsequently in such lesions.     

THE SIGNIFICANCE OF CERTAIN SKIN CHARACTERS OF SHEEP IN RESISTANCE AND SUSCEPTIBILITY TO FLEECE-ROT AND BODY STRIKE

Micro-anatomical differences in skin structure associated with resistance and susceptibility of sheep to fleece-rot and body strike were identified, and found to be of similar magnitude in 2 genetically divergent flocks of medium-woolled Merino ewes. Susceptible sheep were characterised by smaller follicle groups, resulting in higher densities of follicle populations with greater concentrations of the primary follicles (and sudoriferous glands) than in the resistant sheep. From these smaller follicle groups of susceptible sheep, thicker wool fibres grew than were found in the resistant sheep.
The significance of these findings is discussed in relation to the pathogenesis of fleece-rot and body strike of sheep, and the value of the measured skin characters as a method for identifying resistant sheep.     

Immunisation of sheep against experimental Pseudomonas aeruginosa dermatitis and fleece-rot associated body strike

Sheep immunised with an experimental Pseudomonas aeruginosa vaccine and unvaccinated control sheep were challenged by induction of experimental dermatitis with the homologous strain. All of 6 control sheep developed ulcerative dermatitis, and 2 of the 6 challenge sites were struck by larvae of Lucilia cuprina. Neither severe dermatitis nor strike occurred in 6 vaccinated sheep. These results were confirmed in an experimental challenge using 3 different serotypes of P. aeruginosa on each of 3 vaccinated and control sheep, although fly-strikes did not occur. In a field trial of the same vaccine, none of 26 vaccinated sheep developed severe exudative, fleece-rot lesions nor were any fly-struck, whereas 61 of 115 control sheep developed severe, exudative, fleece-rot lesions, 21 of these becoming struck by L. cuprina. The isolates of P. aeruginosa recovered from the field challenge experiment were a different serotype to that used in the vaccine.     

Dermal and serological response against Pseudomonas aeruginosa in sheep bred for resistance and susceptibility to fleece-rot

Genetically select lines of Merino sheep have been bred at Trangie (NSW Agriculture and Fisheries) for resistance (R) or susceptibility (S) to fleece-rot and flystrike. It is believed that fleece characters are primarily responsible for the R or S phenotype. When transferred to the wetter coastal environment of Sydney, R and S sheep with no more than 6 weeks wool cover, continued to show significant differences in the incidence and severity of fleece-rot dermatitis. To test the hypothesis that these sheep might also exhibit differences in their local skin reactions and immune responsiveness, 3 intradermal injections of killed Pseudomonas aeruginosa were administered at monthly intervals. After primary intradermal challenge, R sheep had a higher incidence of skin induration and a stronger inflammatory response (increased induration diameter) than S sheep. Compared to S sheep, R sheep also developed higher levels of circulating antibodies against whole cell antigen and both inner and outer membrane proteins of P. aeruginosa. These responses were maintained in R sheep with each consecutive challenge while S sheep showed a decline in their immune responsiveness. Differences in antibody response against outer membrane proteins were also detected when antigenically naive sheep from each genetic line were sensitised by epicutaneous challenge with P. aeruginosa under experimental wetting conditions. Intradermal challenge of these animals 6 months later with outer membrane proteins, revealed a late maximum (72 h) in the development of induration diameters for R sheep while S animals showed maximal induration diameters by 24 h. However, there was no significant difference in induration response between 24 h and 72 h within each group of sheep.(ABSTRACT TRUNCATED AT 250 WORDS)     

Experimental production of dermatitis in sheep with Pseudomonas aeruginosa

SUMMARY The attachment, to sheep skin, for 4 days, of control wool pads saturated with sterile culture medium which contained a bacteriostat, induced only a mild dermatitis, whereas wool pads saturated with Pseudomonas aeruginosa culture induced a subacute dermatitis characterised by scaling, microabscess formation and seropurulent exudate. Changes similar to the latter were observed in skin affected by natural fleece-rot which developed spontaneously after 7 days of artificial wetting and in which P. aeruginosa was the predominant species of bacteria. An exacerbatory, if not causal, role for this organism is suggested in the development of the dermatitis associated with fleece-rot and in the exudation of seropurulent material, a step essential in the development of body strike.     

OBSERVATIONS ON FIBRE DIAMETER VARIATION OF SHEEP IN RELATION TO FLEECE-ROT AND BODY STRIKE SUSCEPTIBILITY

SUMMARY: A comparative study of various fleece properties known to influence fleece-rot susceptibility was made in a merino flock consisting of sheep which were found to be either resistant or susceptible to fleece-rot and body strike following heavy rains. The fleece properties measured were fibre diameter, fibre diameter variation, wax content, suint content, wax to suint ratio, suint pH, insoluble nitrogen content, wool colour and wettability. Fibre diameter variation, due mainly to the presence of coarse, secondary fibres in the staple, was the only fleece property which differed significantly (p < 0.001) between resistant and susceptible animals. The coefficient of variation of fibre diameter was lowered from a mean value of 22.7 ± 0.3% in susceptible sheep to 20.0 ± 0.3% in resistant sheep. A causal relationship between high fibre diameter variation and fleece-rot susceptibility is suggested. Sheep with irregular fibre size may retain free moisture in the fleece for longer, and thereby become more susceptible to fleece-rot than sheep with uniform fibre diameter, other predisposing factors being equal.     

A morphometric analysis of the changes with age in the skin surface wax and the sebaceous gland area of Merino sheep

SUMMARY Skin samples were collected from the midside of Merino ewes at 6 weeks, one year, and 3 years of age. The thickness of the surface wax and the number and area of sebaceous glands per unit length of skin were measured with an image analyser. Three-year-old sheep had a significantly (P < .05) thicker skin surface wax layer, (14.4μ± 1.45), than they did as lambs, (7.85μ± 0.88). Both the number (2.91 ± 0.15 per mm) and area (24.27 μ²/μ± 1.54) of sebaceous glands per unit length of skin of 3-year-old sheep were significantly less than in lambs, which measured 4.10 ± 0.15 per mm and 39.30 ± 4.14 μ²/μ respectively and one-year-old sheep, which measured 3.99 ± 0.21 per mm and 35.43 μ²/μ 3.65 respectively.     

THE OVIPOSITIONAL RESPONSE OF THE AUSTRALIAN SHEEP BLOWFLY, LUCILIA CUPRINA, TO FLEECE-ROT ODOURS

SUMMARY The ovipositional response of Lucilia cuprina flies to odours emanating from fleece-rot lesions of greasy wool in which Pseudomonas aeruginosa bacteria proliferated, was studied. Fractionation of the fleece-rot odours was carried out by bubbling the volatile components through hydrochloric acid and sodium hydroxide solutions to remove basic odours and acidic odours respectively. It was found that the acidic/neutral odours of fleecerot wool, when perfused into wet, greasy wool, stimulated L. cuprina to oviposit. On the other hand, the basic/neutral odours of fleece-rot wool were virtually unattractive to the gravid fly. Similarly, the acidic/neutral odours emanating from fleece-rot lesions of clean wool from which the non-fibre components, wax, suint and epithelial debris, had been removed by scouring, were found to be unattractive to the gravid fly in choice tests.     

SHH在美利奴羊与小尾寒羊背部皮肤表达差异的研究

为了研究音猬因子(sonic hedgehog, SHH)在不同品种绵羊背部皮肤中的表达差异,以探索SHH与羊毛弯曲形成的关系,选取美利奴羊和小尾寒羊作为研究对象,利用HE染色法观察2种绵羊背部皮肤的组织结构,采用免疫组织化学技术、实时荧光定量PCR和Western blot方法探究2种绵羊背部皮肤中SHH蛋白和基因水平的相对表达量差异。结果显示:HE染色显示美利奴羊与小尾寒羊背部皮肤毛囊的组织结构存在毛囊数量及弯曲程度等差异;免疫组织化学试验显示,SHH蛋白在美利奴羊和小尾寒羊背部皮肤中均有表达,在美利奴羊背部皮肤中的毛乳头、毛基质、内外根鞘、皮脂腺和表皮细胞中显著表达,在小尾寒羊背部皮肤中的毛基质、内外根鞘和皮脂腺中显著表达;光密度分析显示,美利奴羊背部皮肤组织中SHH蛋白的表达量显著高于小尾寒羊(P<0.001);实时荧光定量PCR显示,美利奴羊SHH mRNA相对表达水平显著高于小尾寒羊(P<0.01);Western blot显示,美利奴羊SHH蛋白相对表达水平显著高于小尾寒羊(P<0.01)。提示:羊毛弯曲的形成与其组织结构有关,并受SHH的调控。     

SHH在美利奴羊与小尾寒羊背部皮肤表达差异的研究

为了研究音猬因子(sonic hedgehog, SHH)在不同品种绵羊背部皮肤中的表达差异,以探索SHH与羊毛弯曲形成的关系,选取美利奴羊和小尾寒羊作为研究对象,利用HE染色法观察2种绵羊背部皮肤的组织结构,采用免疫组织化学技术、实时荧光定量PCR和Western blot方法探究2种绵羊背部皮肤中SHH蛋白和基因水平的相对表达量差异。结果显示:HE染色显示美利奴羊与小尾寒羊背部皮肤毛囊的组织结构存在毛囊数量及弯曲程度等差异;免疫组织化学试验显示,SHH蛋白在美利奴羊和小尾寒羊背部皮肤中均有表达,在美利奴羊背部皮肤中的毛乳头、毛基质、内外根鞘、皮脂腺和表皮细胞中显著表达,在小尾寒羊背部皮肤中的毛基质、内外根鞘和皮脂腺中显著表达;光密度分析显示,美利奴羊背部皮肤组织中SHH蛋白的表达量显著高于小尾寒羊(P<0.001);实时荧光定量PCR显示,美利奴羊SHH mRNA相对表达水平显著高于小尾寒羊(P<0.01);Western blot显示,美利奴羊SHH蛋白相对表达水平显著高于小尾寒羊(P<0.01)。提示:羊毛弯曲的形成与其组织结构有关,并受SHH的调控。     

Symmetrical alopecia, scaling and hepatitis in a rabbit

A 5-year-old rabbit with inappetence, symmetrical alopecia and skin lesions was examined. No mites or Malassezia were found in skin scrapings and tape impressions and dermatophyte culture was negative. Trial therapy with ivermectin did not reduce skin lesion severity, and euthanasia was performed because of anorexia after 1 month. Histopathology of the skin showed hyperkeratosis, lymphocytic exocytosis, cell-poor interface dermatitis (lymphocytic infiltration and apoptotic cells in basal layer of epidermis), absence of sebaceous glands and lymphocytic mural folliculitis comparable to sebaceous adenitis and thymoma-associated exfoliative dermatitis previously described in rabbits. The liver exhibited an interface hepatitis, comparable to autoimmune hepatitis in man. The occurrence of morphological similarities to exfoliative dermatitis and sebaceous adenitis in rabbits, in association with an autoimmune hepatitis, has not been described before.     

Immunohistochemical evaluation of substance P and calcitonin gene-related peptide in skin and periosteum after extracorporeal shock wave therapy and radial pressure wave therapy in sheep

OBJECTIVE: To evaluate the effect of focused extracorporeal shock wave therapy (ESWT) and radial pressure wave therapy (RPWT) on immunohistochemical staining for substance P and calcitonin gene-related peptide (CGRP) in the skin and periosteum of sheep. ANIMALS: 36 sheep. PROCEDURES: All 4 limbs of 36 sheep were treated with ESWT, RPWT, or a sham treatment. For 14 days after treatment, at least 2 sheep were euthanized daily and tissue was harvested for histologic evaluation of nerves via staining for substance P and CGRP in the skin and periosteum. RESULTS: No effects of ESWT or RPWT were observed on the number of nerves with stain uptake for substance P or CGRP in the skin or periosteum. CONCLUSIONS AND CLINICAL RELEVANCE: Substance P- and CGRP-containing nerve fibers are not disrupted by EWST or RPWT. Further studies are needed to identify the mechanism of analgesia observed in association with these treatment modalities.     

不同日龄滩羊皮肤组织化学特征比较

为比较2、35日龄滩羊皮肤毛囊的发育特点与血管内皮生长因子（vascular endothelial growth factor，VEGF）和血管内皮生长因子受体2（vascular endothelial growth factor receptor 2，VEGFR2）的分布特征，探究出生后滩羊被毛生长发育的变化特点，试验应用常规HE染色及改良Masson胶原纤维染色、Gomori银氨法染色、磷钨酸染色等特殊染色观察2与35日龄滩羊皮肤组织结构特点；应用免疫组织化学法结合免疫荧光染色法观察VEGF及VEGFR2在2与35日龄滩羊皮肤组织中的分布定位，并用IPP图像分析软件进行定量分析。结果显示：与2日龄滩羊皮肤组织比较，35日龄滩羊表皮与真皮间界限更加清晰，毛囊结构发育完整；毛囊密度显著降低（P<0.05）；胶原纤维与弹性纤维含量增加，形成网格状分布。免疫组化及免疫荧光结果显示，VEGF及VEGFR2在滩羊皮肤表皮及毛囊外根鞘、皮脂腺上均有表达。统计表明，VEGF及VEGFR2在2日龄滩羊皮肤组织中的表达量均显著高于35日龄（P<0.05）。综合上述结果，滩羊毛囊发育过程中，胶原纤维和弹性纤维增加明显；VEGF与VEGFR2通路在毛囊角质形成中起直接调节作用。     

CORYNEBACTERIUM PSEUDOTUBERCULOSIS INFECTION OF SHEEP: ROLE OF SKIN LESIONS AND DIPPING FLUIDS

Caseous lymphadenitis (CLA) infection was established experimentally in a total of 30, 2-tooth merino wethers. In the first experiment a broth culture of the causative organism, Corynebacterum pseudotuberculosis was placed directly on to a freshly shorn area on the right shoulder of 21 sheep after pretreatment with a defatting agent, water or 4 different commercial sheep dips. Within 6–13 days focal skin lesions appeared on 7 sheep and these were followed by abscessation of the right prescapular lymph node. Lung lesions of CLA developed in 6 sheep; 5 of which had skin lesions. In the second experiment C. Pseudotuberculosis broth culture was applied over the right shoulder region of 14 sheep, 24 hours post shearing. The 7 animals had only 1 application of the culture and the remaining 7 had 4 applications on consecutive days. A total of 9 of the 14 sheep developed skin lesions and subsequent CLA in the right prescapular lymph nod but there was no significant difference between the 2 methods of application. As part of the second experiment a broth culture of C. pseudotuberculosis was added to a commercial arsenical sheep dip and 2 hours later sprayed on to the right side of sheep shorn 24 hours or 2 weeks previously. Within 6–13 days all 14 sheep treated in this way developed skin lesions from which C. pseudotuberculiosis was recovered in dense pure growth. Healing time of skin lesions in all affected sheep ranged from 17–72 days post inoculation. A serological test to detect the presence of C. pseudotuberculosis antitoxin was used at regular intervals on all experimental sheep. The earliest positive reaction in an affected sheep was recorded 4 weeks post inoculation. In 23 known affected sheep tested at 35 weeks post inoculation, 10 were positive to the Zaki test. No false positive reactors were recorded. The localisation of focal lung lesions in the interstitial tissue suggests that pulmonary abscessation caused by C. pseudotuberculosis develops via the haematogenous route.     

Efficacy against sheep lice (Bovicola ovis) and fleece wetting of six shower dip preparations

Summary The relative efficacy of 6 shower dip chemicals most frequently used for the treatment of sheep lice (Bovicola ovis) in Western Australia was examined. Groups of 20 sheep infested with lice were treated with products containing either alphamethrin, cyhalothrin, diazinon or diazinon plus piperonyl butoxide and rotenone, formulated as emulsifiable concentrates, and with products containing either coumaphos or magnesium fluorosilicate, formulated as wettable powders. All treatments were applied through a shower dip (Sunbeam model SSD). Inspections for lice were conducted until 9 months after dipping. No lice were found on sheep treated with the 4 emulsifiable concentrate products. In contrast, treatment with the wettable powders, which contained either coumaphos or magnesium fluorosilicate as the active ingredient, did not eradicate the lice infestations. The degree to which the fleece was wetted was assessed 20 minutes after dipping and showed that the wettable powder dips penetrated the fleece less than the emulsifiable concentrate dips. Less fluid was retained by wool staples in an in-vitro test when dip wash was made with the wettable powders. It was concluded that the degree of wetting attained at dipping was an important factor in achieving eradication of sheep lice.     

不同放牧率对草原植物与土壤C、N、P含量的影响

本文研究不同放牧率对草原植物土灌系统中C、N、P含量的影响。结果表明,对照区地上生物量和0～60m土层的总根量显著大于各放牧处理(P＜0.01),而放牧处理间差异不显著(P＞0.05)。对照区和1.33只羊/hm²处理植物的C含量显著大于4只羊/hm²和6.67只羊/hm²,而后两个处理的N、P含量较高。4只羊/hm²处理的C/N和C/P最小。6.67只羊/hm²处理已使根系和根系中的N素明显向表层聚集。对照区0～10cm土层土壤有机C含量显著大于各放牧处理,而各放牧处理间差异则不显著。土壤有机C,N含量与报系生物量成显著正相关。0～10cm土层中无机N在4只羊/hm²处理中最大。土壤微生物量C、P含量在6.67只羊/hm²处理中显著降低,其它处理间差异不显著。其中4只羊和6.67只羊/hm²处理的微生物N含量较高。     

Localization of the candidate genes ELOVL5 and SCD1 for 'male effect' pheromone synthesis in goats (Capra hircus)

The 'male effect' is a well-known phenomenon in female sheep and goats, whereby pheromone-induced activation of reproductive function occurs. In a previous study, we showed that the genes for elongation of long-chain fatty acids family member 5 (ELOVL5) and stearoyl-CoA desaturase 1 (SCD1) increased their expression significantly, concomitant with induction of pheromone synthesis. Therefore, these genes were considered to be prime candidate genes for pheromone synthesis. In the present study, we performed in situ hybridization to investigate where these two genes are expressed in goat skin. Strong positive signals were detected for both genes in the head skin of the male goat, which is the main site of pheromone production, and were mainly in the basal layer of the sebaceous gland cells, with the remaining cells showing negligible signals. None of the cells in the rump skin of the male goat or the head skin of the orchidectomized goat, neither of which produce pheromone, exhibited strong positive signals. The present study demonstrates that expression of these two candidate genes for pheromone synthesis is primarily localized in the sebaceous glands of the pheromone-producing skin region.     

The use of high-energy electrons to depilate the breech of sheep

The use of high-energy electrons for permanently depilating areas of sheep skin was evaluated. The most effective dose of electrons for depilation was 17.5Gy. Histological changes in skin treated at this dose were examined in one sheep over 18 months and in 5 sheep over 89 d. Effects of treatment on bodyweight gains and fleece growth were examined by comparing the productivity of a further 5 sheep treated on the breech with high-energy electrons, with that of conventionally mulesed sheep (n = 5) and untreated controls (n = 5). Electron treatment resulted in immediate death of cells in the germinative region of the wool follicle bulbs. Within 10 d of treatment the treated areas were completely depilated. Wool follicle shafts in the treated areas regressed rapidly towards the epidermis and remained quiescent for the whole trial (89 d). A sheep treated 18 months previously has remained largely depilated, although a few sparse fibres are present. Epidermal acanthosis and orthokeratosis were present at 26 d after treatment. The thickened stratum corneum then sloughed off, but the epidermis remained acanthotic for the entire trial. Sweat glands and most sebaceous glands were destroyed by the treatment and were replaced by fibrotic, avascular tissue in the dermis. In all other respects the external appearance of the electron-treated breech was similar to that of mulesed sheep. There were no apparent side-effects of treatment. Neither mulesing nor electron treatment altered weight gains or fleece growth rates.     

Feeding of the chewing louse Damalinia ovis (Schrank) (Phthiraptera:Trichodectidae) on sheep

The possibility that Damalinia ovis ingests intact epidermis could explain economically important irritation behaviour by infested sheep. To test this, feeding lice were observed on sheep by hand lens and macrophotography and on light and scanning electron microscope preparations. Aspects of feeding behaviour were described, particularly an epidermal ingestion posture. Louse ingesta were examined in paraffin and frozen sections. Lipid-covered stratum corneum squames were identified in crop, midgut, rectum and faeces. Nucleated keratinocytes from inner epidermal strata were not seen, neither were wool fibres. Vertical frozen sheep skin sections, with feeding lice cryofixed by liquid nitrogen, showed mandibles engaged in the outer stratum corneum of the epidermis. The epidermal origin of some ingesta was confirmed by confirming lice over sheep whose skin was stained with Oil Red O, whose colour was seen in crop, midgut, rectum and faeces of harvested lice. Lipase was found in louse midgut. It was concluded that sebaceous secretions may form an important component of the D. ovis diet. The relative contribution from loose scurf or from stratum corneum squames was not determined. Mechanical stimuli by feeding lice did not seem to be the source of host irritation. The role of sensitising agents in louse brei filtrates was investigated with inconclusive results. Intradermal injections of louse faeces did not elicit a demonstrable response.