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.     

AN IN-VITRO TECHNIQUE FOR STUDYING FLEECE-ROT AND FLY STRIKE IN SHEEP

Fleece-rot was experimentally induced in-vitro by wetting and incubating Merino wool samples embedded in serum-agar. Gravid Lucilia cuprina were readily-attracted to these wool culture plates to oviposit. Where serum was freely available to newly-hatched larvae, fly strike and larval development ensued. Using this technique, fleece-rot and oviposition were found to be markedly influenced by the availability of protein and by bacterial activity, particularly that of Pseudomonas spp. The results indicated that odours emanating from wool culture plates containing the latter species played an important role in oviposition. Furthermore, these events varied according to the type of fleece selected, and could be prevented by the addition of a bactericide.     

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)     

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.     

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.     

Patch test reactions to house dust mites in dogs with atopic dermatitis

The purpose of this study was to determine the percentage of dogs with spontaneous atopic dermatitis that show a positive patch test reaction to a commercially available 20% house dust mites mixture containing equal parts of Dermatophagoides farinae and Dermatophagoides pteronyssinus in white petrolatum. In addition, we evaluated whether skin reactions induced after the epicutaneous application of house dust mites were clinically and histologically similar to naturally developed skin lesions of dogs with atopic dermatitis. Furthermore, we investigated if the reactions induced by house dust mites were true allergic reactions by comparing them to atopic lesional skin and to patch test reactions induced by an irritant substance (sodium lauryl sulphate). White petrolatum alone and nonlesional skin sites were used as negative controls. Macroscopic and microscopic evaluations of the patch test and control sites were performed in a blinded fashion at 48 and 72 h after patch test application. Microscopic results were evaluated in a qualitative and quantitative manner. A chi‐square test for homogenicity was used for the quantitative analysis to compare the proportion of each dermal inflammatory cell type among positive histopathological tested sites. P values ≤ 0.05 were considered significant. The study included 12 healthy nonatopic dogs and 13 dogs with nonseasonal atopic dermatitis. None of the nonatopic dogs reacted to house dust mites and white petrolatum. Ten (77%) of the 13 atopic dogs reacted macroscopically and histopathologically to house dust mites. Macroscopic reactions induced by house dust mites were characterized by erythema, oedema and papules. The macroscopic reactions induced by house dust mites were identical to lesional skin in 20% of the dogs and identical to reactions induced by sodium lauryl sulphate in 40% of the dogs. Qualitative histopathological findings showed that the reactions induced by house dust mites were similar to atopic lesional skin in 80% of the dogs and were similar to sodium lauryl sulphate in 20% of the dogs. Quantitative analyses showed that the proportion of neutrophils in reactions induced by sodium lauryl sulphate was significantly higher (P < 0.05) compared to house dust mites reactions, which could be a differentiator factor between an allergic and an irritant reaction. These results showed that the epicutaneous application of house dust mites in dogs with atopic dermatitis induced histopathological lesions similar to spontaneous atopic lesions in dogs. Therefore, this study demonstrated that house dust mites penetrated the skin of dogs with atopic dermatitis and induced an inflammatory response that resembled a true allergic reaction. Funding: Small Companion Animal Grant, University of Minnesota.     

EFFECT OF DISRUPTION OF THE SEBACEOUS LAYER OF THE SHEEP''S SKIN ON THE INCIDENCE OF FLEECE-ROT

It was postulated that integrity of the sebaceous layer on the skin is important in maintaining resistance to fleece-rot. This hypothesis was tested in an experimenta of a 2 x 2 factorial design in which sebaceous layer disruption and wetting were the 2 treatments. The sebaceous layer was disrupted by applying a light petroleum solvent to the skin of the sheep. Fleece-rot occurred in all sheep (20) that were both wetted and had the sebaceous layer disrupted while 9/19 of those wetted but without the sebaceous layer disrupted developed fleece-rot. The fleece-rot lesions observed were more severe in sheep in the disrupted wetted treatments than in the intact wetted treatment. Interpretation of these differences was complicated by a possible reaction of the skin to the solvent used. The differences were not due to an effect of treatment on fleece wettability. No fleece-rot developed in the sheep not wetted and application of the solvent alone did not induce fleece-rot.     

A case of Trichophyton mentagrophytes infection in a fennec fox (Vulpes zerda)

A 2‐year‐old male fennec fox presented with a 4 month history of nonpruritic, crusty skin lesions on the forehead, the pinnae and the tail tip. Initial investigations, including routine haematology, biochemistry profile, multiple skin scrapings, trichoscopic examination, Wood’s lamp examination and fungal culture, failed to reveal any abnormalities. Histopathological examination of a first set of skin biopsies showed an interface dermatitis pattern, with lymphocyte infiltration in the basal layer, a significant lymphocytic exocytosis and occasional apoptotic basal epidermal keratinocytes; periodic acid Schiff stain did not reveal any fungal elements. On further biopsies, there was a pustular neutrophilic dermatitis, with numerous crusts containing high numbers of arthrospores and fungal hyphae. Trichophyton mentagrophytes infection was confirmed on fungal culture and PCR. The fennec fox received oral itraconazole (5 mg/kg once daily for 6 weeks) combined with a miconazole and chlorhexidine shampoo applied on affected areas once weekly, followed with an enilconazole dip. The fox improved dramatically, and a fungal culture performed at 6 weeks was negative. Unfortunately, a few days later the fennec fox developed anorexia, icterus and died. To the authors’ knowledge, this is the first report of Trichophyton infection in a fennec fox and, although a postmortem examination was not performed, this is possibly the first report of fatal acute liver failure associated with itraconazole in a canid.     

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.     

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.     

Combined moxidectin and environmental therapy do not eliminate Chorioptes bovis infestation in heavily feathered horses

Chorioptes bovis infestation is a common cause of pastern dermatitis in the horse, with a predilection in draft horses and other horses with thick hair ‘feathers’ on the distal limbs. The treatment of this superficial mite is challenging; treatment failure and relapse are common. Furthermore, C. bovis infestation may affect the progression of chronic pastern dermatitis (also known as chronic proliferative pastern dermatitis, chronic progressive lymphoedema and dermatitis verrucosa) in draft horses, manifesting with oedema, lichenification and excessive skin folds that can progress to verruciform lesions. An effective cure for C. bovis infestation would therefore be of great clinical value. In a prospective, double‐blind, placebo‐controlled study, the efficacy of oral moxidectin (0.4 mg/kg body weight) given twice with a 3 week interval in combination with environmental treatment with 4‐chloro‐3‐methylphenol and propoxur was tested in 19 heavily feathered horses with clinical pastern dermatitis and C. bovis infestation. Follow‐up examinations over a period of 180 days revealed significantly more skin crusting in the placebo group than in the treatment group. However, no other differences in clinical signs or the numbers of mites detected were found between the two groups. The results of this study suggest that moxidectin in combination with environmental insecticide treatment as used in this study is ineffective in the treatment of C. bovis in feathered horses.     

Induction of feline flea allergy dermatitis and the incidence and histopathological characteristics of concurrent indolent lip ulcers

The objectives of this study were to characterize the role of intermittent vs. continual flea exposure in the development of flea allergy dermatitis (FAD) in cats, assess the accuracy of intradermal skin testing (IDST) and in vitro testing, and document the incidence and histopathological features of indolent lip ulcers. Ten flea‐naive cats were divided into two groups. One group received intermittent flea exposure for 120 days. Thereafter, both groups of cats received continuous flea exposure for 120 days. In vitro testing for flea salivary antibody and IDST utilizing both whole flea antigen and flea salivary antigen were performed. Eight of 10 cats developed clinical signs of FAD within 3 months and five of these eight cats developed lip ulcers which where characterized histopathologically by ulceration with predominantly neutrophilic inflammation and surface bacterial colonization. There was no association between the presence or absence of clinical signs and positive IDST or in vitro results, and no difference in the development of clinical signs was noted between the two groups of cats.     

Dermatomycosis caused by Paranannizziopsis australasiensis in five tuatara (Sphenodon punctatus) and a coastal bearded dragon (Pogona barbata) in a zoological collection in New Zealand

CASE HISTORY: Health monitoring of tuatara (Sphenodon punctatus) at Auckland Zoo between 2001 and 2009 showed that 58/93 tuatara had been affected by dermatitis of unknown origin. From 2011 onwards, cases of suspected fungal dermatitis underwent extensive diagnostic investigations.

CLINCAL FINDINGS: Six cases of dermatomycosis were attributed to Paranannizziopsis australasiensis, five in tuatara and one in a coastal bearded dragon (Pogona barbata). Cases presented typically as raised, yellow to brown encrustations on the skin. Severe cases progressed to necrotising ulcerative dermatitis, and in the bearded dragon to fatal systemic mycosis. Following topical and systemic treatments, lesions resolved in all five tuatara.

LABORATORY FINDINGS: Histopathological examination of skin biopsy samples revealed dermatitis with intralesional septate branching hyphae. Fungal culture yielded isolates morphologically resembling Chrysosporium species, and isolates were submitted for molecular confirmation and sequencing of DNA.

DIAGNOSIS: All six cases were confirmed as dermatitis due to infection with P. australasiensis, on the basis of fungal culture and DNA sequencing of isolates.

CLINICAL RELEVANCE: These are the first reported cases of dermatomycosis associated with P. australasiensis infection in tuatara, and the first cases in which systemic therapeutic agents have been used in the treatment of such disease. Tuatara at the Auckland Zoo are now routinely examined every 3 months and tissue samples from any lesions sent for histopathology and fungal culture. Further work to elucidate the epidemiology and significance of P. australasiensis infections in reptiles in New Zealand is important for both welfare and conservation purposes.     

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.     

P‐87 Alopecia in two guinea pigs due to hypopodes of Acarus farris

Mites are frequent agents of skin diseases in veterinary dermatology, particularly in rodents. The authors report an original case of infestation in two guinea pigs of the same household. Two adult guinea pigs were presented with a 3‐month history of alopecia, matted hair, erythema, scales and mild pruritus. The first guinea pig showed almost complete truncal alopecia with mild scaling, whereas the other one had only recently developed mild alopecia of the abdomen and mild pruritus. Numerous live, long‐legged mites were present both in adhesive tape and skin scrapings. Mycology (Wood's lamp and DTM agar culture) was negative. Histopathological examination of a skin biopsy did not reveal any significant inflammatory disease and morphological changes were limited to hair follicles in the telogen phase. Mites were identified as hypopodes of Acarus farris (Acaridae), but no adult stages were present. Hypopodes were also found in the hay. Control was obtained with acaricidal treatment with Phoxim 0.05% once weekly for 4 weeks, and the elimination of contaminated hay. The lesions and the alopecia resolved rapidly. Hypopodes are additional nymphal stages of some mites of the suborder Astigmata. They develop for dispersing or dormancy under disadvantageous conditions. They can be active or inert, and some can be phoretic on the skin surface or intrafollicular. Although nonparasitic, they may cause dermatitis. Acarus farris have been rarely mentioned in the literature as a cause of dermatitis. This condition could be underdiagnosed or misdiagnosed in veterinary dermatology. Funding: Self‐funded.     

Ectoparasites are the major causes of various types of skin lesions in small ruminants in Ethiopia

Ectoparasites are the major causes of skin lesions in animals. Clinical, skin scraping examination, and histopathological studies were conducted to identify and characterize skin lesions in small ruminants caused by ectoparasites. Mange mites, lice, sheep keds, and ticks were collected from the skin of affected animals for species identification. Skin biopsies were collected from affected part of the skin and fixed in 10% neutral buffered formalin for histopathology. Of 1,000 sheep and 600 goats examined, 815 (81.50%) sheep and 327 (54.5%) goats were infested with one or more types of ectoparasites. Sarcoptes scabiei var ovis, Demodex ovis, Psoroptes ovis, Bovicola ovis, Melophagus ovinus, and Amblyomma variegatum and other tick species were identified from sheep. S. scabiei var caprae, Demodex caprae, Linognathus stenopsis, and A. variegatum and other tick species were identified from goats. Gross skin lesions or defects observed on the skin include stained and ragged wool, loss of wool/hair, nodules, crusts, lichenification, and fissuring. Microscopic evaluation of H and E stained skin sections revealed lesions in the epidermal layer such as hyperkeratosis, acanthosis, and melanin inconsistency on the basal cells of the epidermis. Follicular keratosis, perifolliculitis, frunculosis, perivasculitis, and aggregates of inflammatory cells (of acute and chronic type) with fibrosis were experiential in the dermal layer of the skin. Most of the skin lesions caused by ectoparasites are overlapping. Thus, ectoparasites control program should be executed to reduce skin lesions as skins are the major export commodity of the country.     

Characterization of canine filaggrin: gene structure and protein expression in dog skin

Background – Filaggrin (FLG) is a key protein for skin barrier formation and hydration of the stratum corneum. In humans, a strong association between FLG gene mutations and atopic dermatitis has been reported. Although similar pathogenesis and clinical manifestation have been argued in canine atopic dermatitis, our understanding of canine FLG is limited. Hypothesis/Objectives – The aim of this study was to determine the structure of the canine FLG gene and to raise anti‐dog FLG antibodies, which will be useful to detect FLG protein in dog skin. Methods – The structure of the canine FLG gene was determined by analysing the publicly available canine genome DNA sequence. Polyclonal anti‐dog FLG antibodies were raised based on the canine FLG sequence analysis and used for defining the FLG expression pattern in dog skin by western blotting and immunohistochemistry. Results – Genomic DNA sequence analysis revealed that canine FLG contained four units of repeated sequences corresponding to FLG monomer protein. Western blots probed with anti‐dog FLG monomer detected two bands at 59 and 54 kDa, which were estimated sizes. The results of immunohistochemistry showed that canine FLG was expressed in the stratum granulosum of the epidermis as a granular staining pattern in the cytoplasmic region. Conclusions and clinical importance – This study revealed the unique gene structure of canine FLG that results in production of FLG monomers larger than those of humans or mice. The anti‐dog FLG antibodies raised in this study identified FLG in dog skin. These antibodies will enable us to screen FLG‐deficient dogs with canine atopic dermatitis or ichthyosis.     

Effects of eight vehicles on transdermal lidocaine penetration in sheep skin in vitro

This study investigated the effects of vehicles on penetration and retention of lidocaine applied to sheep skin in vitro. Thoracic skin from two sheep was clipped of wool and stored at −20 °C, until used. Skin samples were defrosted and mounted in Franz‐type diffusion cells, and then one of the following formulations, each saturated with lidocaine, was added: sodium lauryl sulphate (SLS) 0.5% in water, SLS 1% in water, dimethyl sulphoxide (DMSO) 50% in water (wt/wt), DMSO 100%, isopropyl myristate 100% (IPM), water alone, diethylene glycol monoethyl ether (DGME) 50% in water (wt/wt) and DGME 100%. The penetration of lidocaine in each skin sample was measured over 8 h. Significantly greater lidocaine skin concentrations and flux (J_SS) were achieved with the nonaqueous vehicles, DMSO 100% (P < 0.00001 and P < 0.01, respectively), followed by DGME 100% and IPM (P < 0.00001 and P < 0.01, respectively). The lag time (t_lag) for lidocaine penetration in the DMSO 100% vehicle was significantly shorter (P < 0.01) compared with all other vehicles except water. Improved transdermal penetration of lidocaine in the DMSO 100% vehicle was likely due to skin barrier disruption, as determined by differences in pre‐ and post‐treatment transepidermal water loss (TEWL). This study has shown that nonaqueous vehicles enhanced penetration of lidocaine in sheep skin to a greater extent than aqueous vehicles, which has implications for topically applied local anaesthesia in sheep.     

Dual deficiency of copper and cobalt in Hawke's Bay

The comparative efficacy of 13 of the sheep dips currently registered in New Zealand was investigated using sheep infested with the louse Bovicola ovis and carrying wool which was about 10 cm long at the shoulder.

With the exception of one synthetic pyrethroid “pour-on” formulation, all products were able to effect a significant reduction in louse populations, relative to untreated controls, for 37 days after treatment. Only four products proved capable of “eradicating” lice and preventing their re-establishment up to 37 days after treatment.

Variations in manufacturers' recommendations relating to the length of wool at dipping, and mode of application of dips are discussed in relation to the results.