The canine hair cycle - a guide for the assessment of morphological and immunohistochemical criteria

The hair follicle has a lifelong capacity to cycle through recurrent phases of controlled growth (anagen), regression (catagen) and quiescence (telogen), each associated with specific morphological changes. A comprehensive classification scheme is available for mice to distinguish the cycle stages anagen I-VI, catagen I-VIII and telogen. For dogs, such a classification system does not exist, although alopecia associated with hair cycle arrest is common. We applied analogous morphological criteria and various staining techniques to subdivide the canine hair cycle stages to the same extent as has been done in mice. Of all the staining techniques applied, haematoxylin and eosin stain, Sacpic, Masson Fontana and immunohistochemistry for vimentin and laminin proved to be most useful. To evaluate the applicability of our criteria, we investigated skin biopsies from healthy beagle dogs (n=20; biopsies from shoulder and thigh) kept in controlled conditions. From each biopsy, at least 50 hair follicles were assessed. Statistical analysis revealed that 30% of the follicles were in anagen (12% early and 18% late), 8% in catagen (2% early, 5% late and 1% not determinable) and 27% in telogen. Thirty-five per cent of hair follicles could not be assigned to a specific cycle stage because not all follicles within one biopsy were oriented perfectly. In conclusion, this guide will not only be helpful for the investigation of alopecic disorders and possibly their pathogenesis, but may also serve as a basis for research projects in which the comparison of hair cycle stages is essential, e.g. comparative analysis of gene expression patterns.     

Oestrogen receptor evaluation in Pomeranian dogs with hair cycle arrest (alopecia X) on melatonin supplementation

The role of oestrogen receptors in dogs with hair cycle arrest (alopecia X) was investigated by immunohistochemistry. The purpose of this study was to determine if hair regrowth in dogs with hair cycle arrest treated with melatonin was associated with a decrease in follicular oestrogen receptors. Fifteen Pomeranians (excluding intact females) with hair cycle arrest were enrolled. Two biopsies were obtained from alopecic areas of the trunk before and after 3 months on melatonin. Haematoxylin and eosin-stained tissues were examined and oestrogen receptor-alpha was demonstrated immunohistochemically. Common histopathological findings included hyperkeratosis, follicular keratosis, excessive tricholemmal keratinization (flame follicles), thin epidermis, few small anagen bulbs, epidermal pigmentation and melanin aggregates within follicular keratin. Melanin aggregates within basal cells and hair were an occasional finding. After 3 months, 40% (six) dogs had mild to moderate hair regrowth. Biopsies from six dogs showed histological evidence of an increase in anagen hairs and eight dogs had a decrease in epidermal pigmentation. Moderate to marked staining intensity of oestrogen receptor-alpha was noted in all sebaceous gland basal cells, all small hair bulbs and follicular epithelium of telogen hairs. There was no oestrogen receptor-alpha staining of nuclei within the epidermis, apocrine glands or dermal fibroblasts. Large anagen hair bulbs had minimal to no oestrogen receptor staining. Hair regrowth was not associated with a change in oestrogen receptor-alpha staining.     

Influence of inflammation and coat type on oestrogen receptor immunohistochemistry

The purpose of this study was to evaluate oestrogen receptor alpha staining in a variety of breeds and skin conditions. The influence of inflammation and coat type on the presence and intensity of oestrogen receptor alpha staining was evaluated. Approximately 1700 haematoxylin and eosin (H&E)-stained slides of skin biopsies were screened for presence of small hair bulbs. Slides from 94 cases were submitted for oestrogen receptor alpha immunohistochemistry. H&E-stained skin biopsy tissues were examined for inflammation and hair follicle stages. Oestrogen receptor alpha staining characteristics of telogen follicles, flame follicles, large anagen bulbs, small hair bulbs and early anagen hairs (capped bulbs) were recorded. To assess the influence of inflammation and coat type on oestrogen receptor staining of hair follicle types, chi-square tests, Fisher's exact tests and logistic regression models were performed. Slides were classified as inflammatory (65) and noninflammatory (29). There were no statistically significant differences in oestrogen receptor staining when comparing inflammatory to noninflammatory skin biopsies or skin biopsies from dogs with different coat types. A subset of 13 noninflammatory biopsies from alopecic skin was identified. There was a statistically significant increase in the number of flame follicles in this subset as compared to all others. Comparison of oestrogen receptor staining of hair follicle types from these biopsies and all other biopsies revealed a statistically significant increase in the number of mature telogen hair follicles stained in this subset. No statistical difference in staining of early follicle stages was noted. Therefore, the oestrogen receptor is unlikely to be the controlling factor for the transition from telogen to anagen in the dog.     

Alopecia areata in Eringer cows

Alopecia areata is a hair loss disorder in humans, dogs and horses with a suspected autoimmune aetiology targeting anagen hair follicles. Alopecia areata is only sporadically reported in cows. Recently, we observed several cases of suspected alopecia areata in Eringer cows. The aim of this study was to confirm the presumptive diagnosis of alopecia areata and to define the clinical phenotype and histopathological patterns, including characterization of the infiltrating inflammatory cells. Twenty Eringer cows with alopecia and 11 Eringer cows without skin problems were included in this study. Affected cows had either generalized or multifocal alopecia or hypotrichosis. The tail, forehead and distal extremities were usually spared. Punch biopsies were obtained from the centre and margin of alopecic lesions and normal haired skin. Histological examination revealed several alterations in anagen hair bulbs. These included peri‐ and intrabulbar lymphocytic infiltration, peribulbar fibrosis, degenerate matrix cells with clumped melanosomes and pigmentary incontinence. Mild lymphocytic infiltrative mural folliculitis was seen in the inferior segment and isthmus of the hair follicles. Hair shafts were often unpigmented and dysplastic. The large majority of infiltrating lymphocytes were CD3⁺ T cells, whereas only occasional CD20⁺ lymphocytes were present in the peribulbar infiltrate. Our findings confirm the diagnosis of T‐cell‐mediated alopecia areata in these cows. Alopecia areata appears to occur with increased frequency in the Eringer breed, but distinct predisposing factors could not be identified.     

Alopecia in a black Labrador retriever associated with focal sub‐follicular panniculitis and sebaceous adenitis

A 6‐year‐old entire male black Labrador retriever was presented with nonpruritic multicentric, well‐demarcated alopecia of 12‐weeks duration. Skin biopsies from the margins of alopecic regions showed sebaceous adenitis and sub‐follicular panniculitis. Biopsies from alopecic areas showed severe follicular atrophy with residual fibrous tracts, loss of sebaceous glands and lymphohistiocytic panniculitis beneath individual atrophic hair follicle groups. These features differed from previous reports of pilosebaceous diseases of dogs and appeared to extend the spectrum of inflammatory patterns in presumed immune‐mediated adnexal diseases of this species. During the 12‐month follow‐up, there was partial hair regrowth without treatment but alopecia was permanent in the centre of larger lesions.     

The effects of thyroid hormones on the skin of beagle dogs

The effects of hypothyroidism on canine skin were determined by comparing morphologic, morphometric, and hair cycle differences in skin biopsy samples from 3 groups of age- and gender-matched Beagle dogs: (1) euthyroid dogs; (2) dogs made hypothyroid by administration of 131I; and (3) dogs made hypothyroid and maintained in a euthyroid state by treatment with synthetic thyroxine. After 10 months of observation, there was slower regrowth of hair 2 months after clipping in the untreated-hypothyroid dogs. Untreated-hypothyroid dogs had a greater number of follicles in telogen and fewer hair shafts (ie, a greater number of hairless telogen follicles) than did the control group. The control dogs had a greater number of telogen follicles but the same number of hair shafts as the treated-hypothyroid group. Treated-hypothyroid dogs had the greatest number of follicles in the growing stage of the hair cycle (anagen). This study suggests that, at least in Beagles, induced hypothyroidism does not affect the pelage as dramatically as has been described in naturally occurring disease. This is because normal Beagles retain hair shafts in follicles for long periods, and the alopecia of hypothyroidism appears to evolve slowly because of the prolongation of this haired telogen stage. The evaluation of thyroxine-treated hypothyroid dogs demonstrates that thyroid hormone supplementation of Beagle dogs with induced hypothyroidism stimulates hair growth.     

A new type of lesion associated with severe fur damage in Canadian ranch foxes and an investigation of possible causes.

In the silver fox, as in its wild ancestor, the red fox (Vulpes vulpes L.), the annual growing phase (anagen) of guard hair follicles occupies at least four months. Severe damage to the hair coat near the end of this growing period was reported in 1985 on many ranches in New Brunswick and Nova Scotia. A histological analysis of serial sections of skin biopsies showed a marked increase in nuclear aberrations in the hair matrix of anagen guard hair follicles. These nuclear aberrations indicated that cells were undergoing apoptosis, a controlled form of cell death. Tissues from affected and unaffected foxes for histological and toxicological analysis, as well as other data, were obtained during visits to 26 ranches in 1986 and 34 ranches in 1987. Histological sections of the 1987 skin samples showed the mean percentage of nuclear aberrations in 43 unaffected foxes to be 0.08 +/- 0.01 (SEM), while that for 49 affected foxes was 0.51 +/- 0.23. The four foxes with the most severe coat damage also had the highest incidences of guard hair matrix cells with nuclear aberrations, ranging from 20 to 100 times greater than the mean for unaffected foxes. The mitotic index of the hair matrix, which normally remains fairly constant during the hair growth phase, was similar for unaffected and affected foxes (1.83 +/- 0.06 and 1.97 +/- 0.07 respectively). Although our analyses of field data have not established a specific environmental factor associated with increased nuclear aberrations, the possible involvement of toxic agents in follicle damage may warrant further investigation.     

Comparison of hair follicle histology between horses with pituitary pars intermedia dysfunction and excessive hair growth and normal aged horses

Background –  Pituitary pars intermedia dysfunction (PPID) in older equids is commonly recognized by a long hair coat that fails to shed. Objective –  The aim of this study was to compare hair follicle stages in PPID‐affected horses with excessively long hair coats with the stages of normal aged horses (controls) and to compare hair follicle stages in PPID‐affected horses after 6 months of treatment with pergolide mesylate with those of control horses. Animals –  Eight PPID‐affected horses and four normal, age‐matched, control horses. Methods –  Skin biopsies were collected from the neck and rump of PPID‐affected and control horses. A diagnosis of PPID was established based on hair coat changes and supportive overnight dexamethasone suppression test results. Skin biopsies were repeated after 6 months of treatment with pergolide. The number of hair follicles in anagen (A) or telogen (T) was counted for each skin biopsy using transverse sections. Results –  Pretreatment biopsies had a greater percentage of A follicles (neck 96%, rump 95%) and a lower percentage of T follicles (neck 4%, rump 5%) in PPID‐affected horses than in control horses (A, neck 15%, rump 25%; and T, neck 85%, rump 75%). After treatment with pergolide, all PPID‐affected horses had improved shedding, and the percentages of A follicles (neck 69%, rump 70%) and T follicles (neck 31%, rump 30%) were not different from untreated control horses (A, neck 68%, rump 82%; and T, neck 32%, rump 18%). Conclusions –  These findings document that excessive hair growth (hypertrichosis) in PPID‐affected horses is due to persistence of hair follicles in A. Furthermore, treatment with pergolide improved shedding and reduced the percentage of A follicles in PPID‐affected horses.     

Alopecia areata with lymphocytic mural folliculitis affecting the isthmus in a thoroughbred mare

A 13-year-old, thoroughbred mare was presented with an 8-year history of multifocal, generalized, noninflammatory alopecia and a 3-month history of alopecia, erythema and scaling of the white star on the forehead and muzzle. Histopathological examination of biopsy samples from multiple sites on the body (mane, neck, shoulder, flank and gluteal region) showed a subtle lymphocytic inflammatory infiltrate affecting and surrounding the anagen hair bulbs, consistent with a diagnosis of alopecia areata. The biopsy sample from the star on the forehead showed atrophic hair follicles with perifollicular and mural mononuclear folliculitis affecting the isthmus. Immunohistochemical staining with a CD3 marker confirmed the T-lymphocytic origin of the inflammatory infiltrate in all the samples. The concurrent presence of lymphocytic infiltration at the bulbar and isthmic level of the hair follicles in the same horse is unusual. This finding may represent a variation of the histological appearance of alopecia areata.     

Histology of the hair cycle in male beagle dogs

A detailed histologic study of the hair follicles of male Beagle dogs was made to determine changes in microscopic structure associated with the three stages in the hair cycle. Observed changes are that the bulbs of hair follicles in anagen are more closely associated with adipose tissue than they are in telogen. This is due to the deeper penetration of the follicles into the subcutaneous tissue during anagen. The hair germ cells of telogen were presumed to arise from the stratum basale of the matrix cells rather than from the outer root sheath cells. During telogen, the dermal papilla is separated from the club hair, but remains in close proximity to it. There is no connecting stalk as is reported for other animals.     

Congenital alopecia in a Bichon Frise

A Bichon Frise pup had congenital alopecia. Histologic evaluation revealed the absence of hair follicles, arector pili muscles, sebaceous glands, and sweat glands. Unlike previously described cases of congenital ectodermal defect, this alopecia was not associated with any color pattern; the pup was white until it was 4 months old, at which time normal black and brown pigmentation developed independently of the alopecic pattern.     

Alkaline phosphatase reaction in hair follicles of male beagle dogs during hair cycle stages.

The literature reviewed has revealed differences of opinion concerning the location of alkaline phosphates and its relationship to the hair cycle stages. This project was undertaken to determine alkaline phosphatase activity in the hair follicle stages: anagen, catagen, and telogen. Skin biopsy samples were collected over a period of 1 year from 9 purebred male Beagle dogs. At the beginning of the experiment 3 dogs were 2 weeks of age, 3 dogs were 12 months of age and 3 dogs were 21 months of age. The skin biopsies were taken monthly from alternate sides of each animal near the thoracolumbar region. Samples were fixed in 80% chilled alcohol and the alkaline phosphatase activity was determined by Gomori's calcium cobalt method. A total of 250 slides with seven sections per slide was examined. Each slide contained between 14–60 hair follicles. During anagen stage, the reaction of the dermal papilla for the presence of alkaline phosphatase was variable. A weak or negative reaction was observed in this study which was an unexpected finding. The reaction of the dermal papilla to alkaline phosphatase was positive in catagen stage. The cells of the dermal papillae in telogen stage were strongly alkaline phosphatase positive. The reaction was not confined to the blood vessels but occurred in all parts of the dermal papilla in both early and late stages.     

Congenital hypotrichosis in a French bulldog

The occurrence of congenital alopecia in one of a litter of four female French bulldogs is described. With the exception of the hair coat the affected bitch was clinically normal. The distribution of the sparse hair (head, tip of the tail and paws) mimicked that seen in the Chinese crested dog. Histopathological examination of skin biopsies revealed features frequently observed in congenital alopecia - a reduction in number of hair follicles, hypoplasia or absence of epidermal appendages and pronounced follicular hyperkeratosis, abnormalities of collagen and elastic fibres were not detected.     

Alopecia areata (pelade) in a cow

A case of alopecia areata (pelade) is described in a cow. Alopecia areata is a rare idiopathic dermatosis of cattle. It is characterized by asymptomatic, solitary or multiple, annular areas of noninflammatory alopecia. Scrapings and cultures are negative, and the diagnosis is confirmed by skin biopsies taken from early lesions, which reveal accumulations of lymphocytes around the bulbs of anagen hair follicles. There is no known effective and practical treatment for affected cattle. The dermatitis is a cosmetic problem, but does not seem to affect general health and production, and spontaneous remission may occur.     

Hair cycle in dogs with different hair types in a tropical region of Brazil

Hair cycle activity has been extensively studied in humans, sheep and laboratory animals, but there is a lack of information in dogs. Besides varying according to species, breed, sex and general health, hair growth is mainly affected by climatic variations. The aim of the study was to evaluate the follicle activity in three breeds of dogs with different hair types, in the city of Viçosa, Minas Gerais (latitude 20°45′S), Brazil. Twenty‐one male dogs of boxer, labrador and schnauzer breeds were trichographically analysed monthly over 12 consecutive months. Hair percentage of telogen and anagen hairs at the different stages of the hair cycle in boxers and labradors was not significantly different, but both differed from the schnauzers. A significant correlation between hair follicle cycle and environmental temperature and photoperiod was noted in the boxers and labradors. In these breeds, a larger number of telogen hairs were observed during the hottest months of the year, and an increase in anagen hairs during the coldest months. The mean percentage of telogen hairs was 93, 90 and 55.3% for boxer, labrador and schnauzer, respectively.     

Mutually-induced traumatic alopecia responsive to husbandry modification in two cohabiting donkeys

Cutaneous expression of behavioural processes remains unexplored in donkeys. This case report describes a case of traumatic alopecia in two donkeys secondary to mutual hair plucking behaviour. Two genetically unrelated, male castrated, 4-year-old Cotentin donkeys living together were presented for a 2-year history of alopecic, mainly nummular, nonpruritic lesions developing on the back, loin and croup. The general physical examination and behavioural assessment were unremarkable. Trichoscopy, scrapings and cytology were performed. Superficial bacterial infection was diagnosed but a 2-week course of antibacterial treatment showed no improvement. Histopathological examination showed features compatible with traumatic alopecia and consistent with several behavioural causes of alopecia resembling human trichotillomania or acute traction alopecia, canine traction alopecia, feline self-induced alopecia and murine barbering. Separation of the donkeys led to complete long-term remission. Clinical and histopathological findings in our case share lots of similar features with the different mechanical traumatic alopecic disorders mentioned above, making it impossible to simply classify the described affection. Whether alopecic lesions resulted from an exclusive behavioural process (either abnormal excessive mutual grooming or other social interactions) or were associated with underlying causes of pruritus is uncertain. Mechanical alopecia secondary to a behavioural process should therefore be suspected in cases of permanent patchy alopecic nummular lesions on donkeys living in groups.     

Oestrogen receptor antagonist and hair regrowth in dogs with hair cycle arrest (alopecia X)

An oestrogen receptor pathway that regulates the telogen-anagen hair follicle transition in mice has been described. The purpose of this study was to investigate whether fulvestrant, a pure oestrogen receptor antagonist, would cause hair regrowth in Pomeranian dogs with hair cycle arrest (alopecia X). Eleven Pomeranian dogs with hair cycle arrest were randomly assigned to receive two intramuscular injections of either 10 mg kg(-1) fulvestrant (n = 6) or an equal volume of saline (n = 5) 1 month apart. Complete blood count, chemistry panel, and urinalysis were monitored prior to the first injection and monthly for 2 months. Dogs were evaluated each month for degree of hair growth, percentage of body affected, and quality of new hair growth. Three control dogs received fulvestrant after the completion of the study. In addition, one control dog and one treatment dog received two subcutaneous injections of 20 mg kg(-1) fulvestrant 1 month apart. No dogs that received 10 mg kg(-1) fulvestrant had any evidence of hair regrowth. The control dog that received 20 mg kg(-1) fulvestrant had substantial hair regrowth 1 month after the first injection. No adverse effects from the treatment were noted. Fulvestrant does not appear to be a feasible treatment for dogs with hair cycle arrest (alopecia X) when administered intramuscularly at 10 mg kg(-1). A higher dose of fulvestrant requires more investigation but may be cost-prohibitive.     

The effects of thyroxine on hair growth in the dog

The effects of orally and topically administered thyroxine on hair growth were studied in normal, intact, cross-bred male dogs maintained under the same controlled conditions of temperature and photoperiod. It was found that, irrespective of the route of administration, thyroxine produced an increase in both the rate of hair growth and in the number of hair follicles entering the anagen phase of the hair cycle. With orally administered thyroxine, the changes in hair growth were most marked in the flank region.     

Resolution of paraneoplastic alopecia following surgical removal of a pancreatic carcinoma in a cat

A 13-year-old female neutered domestic longhaired cat was presented with a five-month history of progressive weight loss and bilaterally symmetrical alopecia of the ventrum, limbs and perineum. The alopecic skin had a shiny appearance and hair in the non-alopecic areas was easily epilated. Fine needle aspirate cytology of a palpable cranial abdominal mass revealed it to be of epithelial or glandular origin. A pancreatic mass was excised by left pancreatectomy during exploratory laparotomy, and histopathology and skin biopsies confirmed a diagnosis of pancreatic carcinoma with concurrent paraneoplastic alopecia. No evidence of metastases was found on liver and lymph node biopsies. At re-examination 10 weeks after surgery, the hair had fully regrown. Skin signs recurred after 18 weeks and metastatic spread of the tumour was confirmed on postmortem examination. This case confirms that paraneoplastic alopecia associated with internal malignancies is a potentially reversible process if the internal neoplasm is excised.