Status of canine demodicosis by using different diagnostic methods and its correlation with endoparasitic infections in dogs of Mumbai and thane region of Maharashtra state

Introduction

In today's veterinary field, small animal dermatology practice has become an important aspect. The skin, being the largest organ of the body, often provides information to the clinician that reflects both local and systemic changes, providing valuable clues for early diagnosis (Barlaskar et al., 2023; Marchegiani et al., 2020) [5,15] . In canines, these changes are an important indicator as they reflect the general health of an animal. Over the past decade, veterinary clinics have seen a noticeable rise in dermatological consultations due to increased owner awareness and the growing prevalence of skin-related diseases. Dermatitis, defined as inflammation of the skin, may be produced by a myriad of agents, including parasites, bacteria, fungi, external irritants, burns, and allergens The contribution of ectoparasites viz. ticks, fleas, lice and mites causing dermatitis is conspicuous. One of the causes of parasitic dermatitis in dogs is the follicular mite, Demodex canis or 'red mange'. Canine demodicosis is a refractory skin disease caused by the excessive Proliferation of demodectic mites in the animal's hair follicles and sebaceous glands (Kumari, 2017) [12] . Clinically, affected dogs may show lethargy, reduced appetite, anaemia, and secondary bacterial infections, especially when additional endoparasitic or ectoparasitic burdens are present (Abel et al., 2023; Sousa et al., 2020) [1,39] . Severe or chronic cases may require months of therapy before full recovery is achieved. Although Demodex mites are considered to be commensals on dog skin, disease develops when the immune system fails to regulate their numbers, allowing unchecked multiplication. Prompt and accurate diagnosis is crucial because inappropriate treatment can worsen the condition (Silva et al., 2021) [33] Corticosteroids, for instance, may be useful in sarcoptic mange but can aggravate demodicosis by promoting mite proliferation (Almeida et al., 2021; Swe et al., 2014) [2,42] . Tentative diagnosis is usually based on the presence of signs such as alopecia, erythema, crusting, pruritus, follicular pustules, furunculosis, ulceration, malodour, and, in cases of pododemodicosis, marked interdigital swelling (Sharma, et al. 2018; Sousa et al., 2020) [29,31,39] . Secondary bacterial infections are also common in advanced disease Deep skin scraping remains the most widely used diagnostic technique and is regarded as the gold standard, as the finding of even a single mite confirms demodicosis (Mueller et al., 2020) [16] . However, it is invasive, uncomfortable for the animal, may cause capillary bleeding, and is often poorly tolerated by both dogs and their owners (Singh, 2019) [35] . In addition, the procedure requires sample processing, which can delay diagnosis (Suartha et al., 2018) [41] . Other methods such as acetate tape impressions and trichograms are available, but they generally offer lower sensitivity or are better suited for generalized lesions (Madureira and Brum, 2017; Mueller et al., 2020) [13,16] . Superglue Slide Impressions (SSI), or cyanoacrylate skin surface biopsies, offer a less invasive, quick, and economical alternative. Well-established in human dermatology for identifying Demodex spp. and Sarcoptes scabiei, this method has recently been adapted for use in dogs. In this technique, cyanoacrylate adhesive is applied to the skin to extract mites, which can subsequently be examined directly under a microscope without additional processing. SSI also allows collection of viable mites for monitoring the effectiveness of treatment (Singh, 2019) [35] . Given the clinical relevance of demodicosis and the limitations of existing diagnostic approaches, the present study was designed with the following objectives From all suspected cases, deep skin scrapings, Superglue Slide Impressions (SSI), faecal samples, and blood samples were collected. Deep skin scrapings were obtained by scraping the skin with a sterile blade dipped in liquid paraffin until capillary oozing of blood was observed. The samples were processed according to the method described by Soulsby (1982) [40] . The scraping was mixed with 5-10 ml of 10% KOH, then heated for 10-15 minutes. After discarding the supernatant, 2-3 drops of sediment were mounted on a slide with a coverslip and examined under low power for the presence of mites SSI samples were collected according to the technique described by Singh (2019) [35] . A drop of cyanoacrylate adhesive was placed on the centre of a clean glass slide; the affected skin was gently squeezed between the thumb and index finger, and the slide was pressed to the area for approximately 30 seconds. The slide was then removed against the direction of hair growth. A drop of immersion oil was added to the impression and was covered with another slide and examined under low magnification for live mites (Fig. 3,4). Mite Index for Demodex spp. was calculated by counting all life stages of Demodex canis (eggs, larvae, nymphs, and adults) in the ratio of immature to mature stages.

$Mite Index=\frac{Number of immature mites}{Number of mature mites}$ Faecal samples from suspected or confirmed demodicosis cases were examined for concurrent endoparasitic infection using the sedimentation method (Soulsby, 1982) [40] . Approximately 2 g of faeces was mixed with 10 ml of water and strained through cotton gauze. The filtrate was transferred into a sedimentation flask, topped with water, and left undisturbed for 30 minutes. After decanting the supernatant, a drop of the sediment was placed on a slide and examined under 10× magnification for parasite eggs or larvae. Peripheral blood was collected from the cephalic vein of dogs confirmed positive for Demodex spp. for haematological analysis. Parameters evaluated included haemoglobin concentration, packed cell volume (PCV), erythrocyte indices, Total Leukocyte Count (TLC), and Differential Leukocyte Count (DLC). Thin blood smears were also prepared to screen for haemoprotozoan parasites, following the method described by Rathore and Sengar (2005) [21] . All data were analyzed statistically following the methods of Snedecor and Cochran (1994) [36] .

Results

Prevalence of canine demodicosis

During the study period, 143 dogs with clinical signs of dermatitis such as alopecia, pruritus, and erythema were examined for demodicosis using deep skin scraping and Superglue Slide Impressions (SSI). Of these, 50 dogs were confirmed positive for canine demodicosis, resulting in an overall prevalence of 34.96%.

3.1.1

Clinical forms of canine demodicosis Canine demodicosis was categorised into localised and generalised forms based on lesion distribution. Among the 50 affected dogs, 9 (18%) had localised demodicosis and 41 (82%) showed generalised involvement

3.1.2

Age-wise prevalence Based on the age of the dog, demodicosis can be classified as juvenile onset or adult onset demodicosis. Out of 50 dogs positive for canine demodicosis, nine dogs (18%) showed juvenile onset, while 41 dogs (82%) had adult onset of the disease (Table 1). The difference between both groups was statistically highly significant (p<0.05).

Breed-wise prevalence

Examined dogs were broadly classified as pedigree or nondescript. Among the 50 demodectic dogs, non-descript dogs accounted for 60% of cases, while pedigree dogs constituted 40%. Within pedigree dogs, Labradors (14%) were most affected, followed by Pomeranian (6%), American Bulldog, Pug, and Shih Tzu (4% each), and Doberman, Lhasa Apso, Pitbull, and Rottweiler (2% each).Statistical analysis showed no significant difference in prevalence between pedigree and non-descript dogs (p>0.05).

Sex-wise prevalence

Of the 50 affected dogs, 31 (62%) were males and 19 (38%) were females. Although males were numerically more affected, the difference between sexes was statistically nonsignificant (p>0.05).

Clinical findings

Dogs with demodicosis showed a variety of dermatological lesions and systemic signs. The most common lesions observed were alopecia (100%), followed by pruritus (76%), pustules (62%), crusting (56%), erythema (52%), scaling and lichenification (48% each), ulceration (26%), comedones (24%), pododemodicosis (24%), and foul odour (22%). (Table 2, Fig. 5678)

Distribution of lesions

Lesions were either confined to specific regions or distributed over multiple body sites. Generalized demodicosis, defined as involvement of five or more body regions, was noted in 41 dogs (82%).The most frequently affected areas were the head and dorsum (68% each), followed by hindlimbs (62%), forelimbs (58%), neck (56%), abdomen (50%), and tail (24%) (Table 3).

Diagnosis of canine demodicosis

Diagnosis was based on deep skin scraping and Superglue Slide Impressions (SSI). Out of the 50 affected dogs, skin scraping detected 47 positives (94% sensitivity), whereas SSI detected 48 positives (96% sensitivity)

Skin scraping

Deep skin scraping (DSS) detected Demodex canis in 47 of 50 dogs (94%). All developmental stages such as eggs, larvae, nymphs, and adults were observed (Fig. 9) Concurrent Sarcoptes spp. infection was detected in 3 dogs (6%), while no cases of Otodectes cynotis infestation were recorded. (Fig. 10)

Superglue slide impressions (SSI)

Using SSI, 48 of 50 dogs (96%) were found positive for D. canis. All life stages of the mite were observed, and live mites were frequently detected, indicating that SSI can also be used to collect viable mites for assessing miticidal efficacy (Fig. 1112). No other mange mites were detected on SSI in the present study.

Comparison of mite counts

Total mite counts (adult and immature stages) from both tests are presented in Table 4. Although numerical differences were seen between methods, the differences were statistically non-significant (p>0.05), indicating comparable quantitative performance. Qualitatively, SSI was easier to perform, required less time, and was more acceptable to both dogs and owners, and no adverse reaction to cyanoacrylate adhesive was observed.

Mite index

The mite index was calculated as the ratio of immature to mature stages of D. canis in each case. Among the 50 dogs, 39 (78%) had a mite index greater than 1, indicating a predominance of immature stages and suggesting an active, progressive infection. The remaining 11 dogs (22%) had mite indices ≤1, consistent with recovering or less active infections.

Concurrent infections 3.4.1 Concurrent ectoparasitic infestation

Out of 50 demodectic dogs, 8 (16%) had flea infestation, 6 (12%) had ticks, 3 (6%) had sarcoptic mange, and 3 (6%) had maggot-infested wounds (Table 5). These concurrent infestations were associated with additional skin irritation, self-trauma, and hypersensitivity reactions.

Concurrent endoparasitic infection

Fecal examination of the 50 demodectic dogs revealed endoparasitic infection in 14 dogs (28%) (Table 6). Hookworm eggs were detected in 6 dogs (12%), Dipylidium caninum egg capsules in 4 dogs (8%), and mixed infections with both hookworm and D. caninum in 4 dogs (8%). A higher proportion of endoparasitic infections was seen in non-descript dogs compared to pedigree dogs.

Blood smear examination

All 50 demodectic dogs were negative for haemoprotozoan parasites on blood smear examination.

Hormonal imbalance

Serum thyroxine evaluation revealed hypothyroidism in 6% of the demodectic dogs. These findings suggest that thyroid dysfunction may be a contributing factor in a subset of cases.

Haematological findings

Haematological values of demodectic dogs and corresponding reference ranges are summarized in Table 7.

Haemoglobin and packed cell volume

Haemoglobin values ranged from 7.1 to 13.3 g%, with a mean of 10.27±0.22 g/dL, indicating a mild to moderate reduction compared to the reference range. Packed cell volume (PCV) ranged from 20.1 to 38.76%, with a mean of 30.13±0.63%, also lower than normal reference values.

Erythrocyte indices

The mean MCV, MCH, and MCHC were 65.88±1.86 fl, 22.52±0.70 pg, and 34.2±0.36 g/dl, respectively, which are compatible with normocytic, normochromic anaemia.

Total and differential leucocyte counts

Total leucocyte counts were elevated, ranging from 13 to 56.6 × 10³/µL, with a mean of 24.73±1.12 × 10³/µL. Differential counts showed neutrophilia (84.2±0.70%), lymphocytopenia (13.44±0.67%), eosinopenia (1.26±0.22%), and monocytopenia (1.2±0.06%).

Discussion

The overall prevalence of canine demodicosis (34.96%) recorded in this study aligns with earlier reports from Maharashtra and Jammu where similar levels of occurrence have been described (Pawar, 2021 and Berian et al., 2018) [4,18] . Conversely, much lower prevalence values ranging from 2.7% to 6% have been observed in other regions of India, including Eastern Maharashtra and Andhra Pradesh (Raut et al., 2006 and Sowjanya, 2019) [14,22] , while Sarma et al. (2013) [27] reported a substantially higher prevalence of 51.92%. These discrepancies likely reflect differences in climate, seasonal conditions, owner awareness, management practices, and diagnostic sampling strategies Generalised demodicosis was significantly more common than the localised form in the present study, a pattern consistent with findings from Assam, Tirupathi and Himachal Pradesh (Pradhan, 2016; Kumari, 2017; Sharma et al., 2018) [12,20,29,31] . Owners may overlook mild localised lesions or delay veterinary consultation, resulting in the presentation of more advanced cases. In contrast, studies from Gujarat and Kerala have reported more localised cases (Nair, 2019; Solanki, 2007) [17,37] possibly due to spontaneous resolution of mild disease. Adult-onset demodicosis predominated in this investigation, a finding consistent with reports from Gujarat (Solanki, 2007) [37] and Himachal Pradesh (Sharma et al., 2018) [29,31] . Adult-onset disease may result from relapse of juvenile demodicosis or from underlying triggers such as chemotherapy, hormonal imbalance, endoparasitism, or compromised immunity (Kumar et al., 2018) [11] . Conversely, Gunaseelan, 2011 [8] reported higher susceptibility in juveniles, likely reflecting immature immunity and close neonatal contact with carrier dams. Nondescript dogs were more frequently affected, probably due to poorer nutrition, inadequate housing, and delayed veterinary care consistent with observations in Palampur (Sharma et al., 2018) [29,31] . On the other hand, a higher prevalence among purebred dogs has also been documented (Nair, 2019) [17] , suggesting breed predisposition and possible hereditary immune defects (Pradhan, 2016) [20] . Although more males were affected, the difference was not statistically significant. Testosteronerelated immunomodulation has been suggested as a factor (Klein, 2004 and Roberts et al., 2004) [10,24] , but many studies, including those from Gujarat and Assam, also reported no meaningful sex association (Pradhan, 2016; Solanki, 2007) [20,37] The clinical signs noted were alopecia, pruritus, pustules, crusting, erythema, scaling, lichenification, ulceration, comedones, pododemodicosis, and foul odour, which reflect classical presentations described across India (Chakraborty, 2012; Sivajothi et al., 2015) [7,34] . These manifestations result from folliculitis, inflammation and secondary bacterial infection caused by Demodex proliferation. The distribution of lesions on the head, dorsum, limbs and neck mirror trends (Sood et al., 2012) [38] . Deep skin scraping remained highly sensitive (94%) and detected all life stages of the mite, reinforcing its role as a standard diagnostic tool. However, Superglue Slide Impressions (SSI) demonstrated slightly higher sensitivity (96%) and have several advantages like faster performance, less invasiveness, higher owner acceptability, and the ability to recover viable mites. Comparable findings were reported in humans using cyanoacrylate biopsies (Bunyaratavej et al., 2016; Zorbozan et al., 2020) [6,44] , and by Singh (2019) [35] in dogs. Mite counts between both methods did not differ significantly, confirming the quantitative reliability of SSI.

Concurrent flea infestation, tick infestation, sarcoptic mange and maggot wounds were relatively common, and similar coinfections have been noted in Nepal (Sharma and Pokharel, 2019) [30] . These additional skin irritants are likely to aggravate pruritus, self-trauma, and immune dysregulation, contributing to worsening disease. Endoparasitic infections were observed in 28% of demodectic dogs, mainly hookworms and Dipylidium caninum. Endoparasites are recognized contributors to immunosuppression and demodicosis (Shaw and Foster, 2000) [32] , and juvenile demodicosis has been linked to hookworms and coccidia (Plant et al., 2011) [19] . Higher prevalence in earlier studies from West Bengal (Chatterjee, 2007; Chakraborty, 2012) [7] may reflect differences in deworming practices. Hypothyroidism (6%) also contributed in some dogs, as low thyroxine is known to impair cell-mediated immunity and facilitate mite proliferation (Sakhare, 2017) [25] . Haematological changes showed mild to moderate anaemia, low PCV, and normocytic-normochromic indices supporting the presence of anaemia of chronic disease, which is commonly reported in demodicosis (Sakhare, 2017; Sharma et al., 2018) [25,29,31] . Leucocytosis and pronounced neutrophilia indicate chronic antigenic stimulation and secondary bacterial infection, as previously documented (Reddy et al., 2015; Boda, 2016) [23] . Lymphocytopenia aligns with earlier suggestions of T-cell suppression (Scott et al., 1974; Sakina et al., 2012) [26,28] . Eosinopenia, although counterintuitive, has been reported internationally (Hillier & Desch, 2002) [9] and may reflect stress-mediated glucocorticoid release (Benjamin, 1978) [3] . Monocytopenia, although observed, is not clinically significant (Valenciano et al., 2014) [43]

References

1. Abel RN, Soma IG, Batan IW. Demodicosis with ticks infestation and babesiosis in a mixed dog. Vet Sci Med J. 2023;5(11). doi:10.24843/vsmj.2023.v5.i11.p05.
2. Pereira Salça de Almeida G, Ribeiro Campos D, Barbour Scott F, Ribeiro Correia T. Successful treatment of feline demodicosis with oral sarolaner: case report. Braz J Vet Med. 2021. https://bjvm.org.br/BJVM/article/view/1134
3. Benjamin MM. Outline of veterinary clinical pathology. 3rd ed. Ames: Iowa State University Press; 1978.
4. Berian S, Gupta SK, Sharma V, Bhardwaj RK, Ali S. Prevalence of canine parasitic dermatitis in and around Jammu. Int J Curr Microbiol Appl Sci. 2018;7(8):24202426.
5. Barlaskar SA, Pradhan S, Mondal S, Mukhopadhayay SK, Hoque R, Sri CPS, et al. Prevalence of dermatophytosis in canines in and around Kolkata metropolitan area. J Anim Res. 2023;13(5):797-807. doi:10.30954/2277-940X.05.2023.22
6. Bunyaratavej S, Rujitharanawong C, Kasemsarn P, Boonchai W, Muanprasert C, Matthapan L, et al. Skin scrapings versus standardized skin surface biopsy to detect Demodex mites in patients with facial erythema of uncertain cause: a comparative study. Indian J Dermatol Venereol Leprol. 2016;82:519-522.
7. Chakraborty S. Dermatitis in dogs with special reference to demodicosis [M.V.Sc. thesis]. Kolkata: West Bengal University of Animal and Fishery Sciences; 2012.
8. Gunaseelan L, Bhavya S, Senthil Kumar K, Balachandran C. Influencing factors for mange mite infestation of dogs in Chennai city. Tamil Nadu J Vet Anim Sci. 2011;7(5):247-249.
9. Hillier A, Desch CE. Dermatoses associated with ectoparasites in dogs. Compend Contin Educ Pract Vet. 2002;24(9):680-692.
10. Klein SL. Hormonal and immunological mechanisms mediating sex differences in parasite infection. Parasite Immunol. 2004;26:247-264.
11. Kumar A, Das AK, Sinha M, Arya SK, Kumar A, Kumar B. Study on the prevalence of demodectic mange in dogs in and around Patna. Int J Curr Microbiol Appl Sci. 2018;7:4216-4221.
12. Kumari DB. Clinico-diagnostic and therapeutic studies on demodicosis in dogs [M.V.Sc. thesis]. Tirupati: Sri Venkateswara Veterinary University; 2017.
13. Madureira R, Brum JS. Dermatological diagnosis in small animals: what can influence? Arch Vet Sci. 2017;22(4). doi:10.5380/avs.v22i4.56944
14. Sowjanya M. Epidemiological surveillance of ectoparasitic infections in dogs in Krishna-Godavari zone of Andhra Pradesh [M.V.Sc. thesis]. Tirupati: Sri Venkateswara Veterinary University; 2019.
15. Marchegiani A, Fruganti A, Spaterna A, Dalle Vedove E, Bachetti B, Massimini M, et al. Impact of nutritional supplementation on canine dermatological disorders. Vet Sci. 2020;7(2):38. doi:10.3390/vetsci7020038
16. Mueller RS, Rosenkrantz W, Bensignor E, Karaś-Tęcza J, Paterson T, Shipstone MA. Diagnosis and treatment of demodicosis in dogs and cats. Vet Dermatol. 2020;31:4e2. doi:10.1111/vde.12806
17. Nair AS. Diagnostic and therapeutic evaluations of demodicosis in dogs [M.V.Sc. thesis]. Wayanad: Kerala Veterinary and Animal Sciences University; 2019.
18. Pawar MU, Sakhare MP, Shafi TA, Siddiqui MFMF, Chigure GM, Gayal SD. Prevalence of canine demodicosis in and around Parbhani. Pharma Innov J. 2021;10(12):548-550.
19. Plant JD, Lund EM, Yang M. A case-control study of the risk factors for canine juvenile-onset generalized demodicosis in the USA. Vet Dermatol. 2011;22(1):9599. doi:10.1111/j.1365-3164.2010.00922.x
20. Pradhan D. Current trend of demodicosis in dog: epidemiology, haemato-biochemical changes and therapeutic management [M.V.Sc. thesis]. Guwahati: Assam Agricultural University; 2016.
21. Rathore RS, Sengar GS. Diagnostic parasitology. Jaipur: Pointer Publishers; 2005. p.156.
22. Raut PA, Maske DK, Jayraw AK, Sonkusale VG. Ectoparasitism in dogs from the eastern zone of Maharashtra state. J Parasit Dis. 2006;30(2):138-141.
23. Reddy BS, Kumari KN, Sivajothi S. Haematobiochemical findings and thyroxin levels in canine demodicosis. Comp Clin Pathol. 2015;24(2):287-290.
24. Roberts ML, Buchanan KL, Evans MR. Testing the immunocompetence handicap hypothesis: a review of the evidence. Anim Behav. 2004;68(2):227-239. doi:10.1016/j.anbehav.2004.05.001
25. Sakhare MP. Therapeutic management of canine demodicosis with immunomodulators [PhD thesis]. Nagpur: Maharashtra Animal and Fishery Sciences University; 2017.
26. Sakina A, Mandial RK, Mudasir Q. Haematobiochemical changes in canine demodicosis. Vetscan. 2012;7:75-77.
27. Sarma K, Mondal DB, Sarvanan M, Kumar M, Vijaykumar H. Incidence of dermatological disorders and its therapeutic management in canines. Intas Polivet. 2013;14(2):186-192.
28. Scott DW, Farrow BRH, Shultz RD. Studies on therapeutic and immunologic aspects of generalized demodicosis in dogs. J Am Vet Med Assoc. 1974;207(10):1308-1310.
29. Sharma P, Wadhwa DR, Katoch A, Sharma A. Epidemiological, clinico-haematological and therapeutic studies on canine demodicosis. J Dairy Vet Anim Res. 2018;7(3):109-113. doi:10.15406/jdvar.2018.07.00200
30. Sharma S, Pokharel S. Diagnosis and therapeutic management of mixed Demodex and Sarcoptes mite infestation in dog. Acta Sci Agric. 2019;3(6):163-166.
31. Sharma K, Sharma R, Gupta S. Haematological alterations in dogs with demodicosis. Himachal J Vet Sci. 2018;7(2):32-37.
32. Silva TRM, Schimmunech MS, Pott PJK, AlvesSobrinho AV, Oliveira PG, Amaral AVC, et al. Mange skin diseases in dogs and cats in Jataí, Brazil: a retrospective study with notes on zoonotic aspects. Res Soc Dev. 2021;10(1):e10610111417. doi:10.33448/rsdv10i1.11417
33. Sivajothi S, Sudhakara Reddy B, Rayulu VC. Demodicosis caused by Demodex canis and Demodex cornei in dogs. J Parasit Dis. 2015;39:673-676. doi:10.1007/s12639-013-0405-3
34. Singh SK. Superglue slide impression (SSI) method: a novel diagnostic application for canine demodicosis. Exp Appl Acarol. 2019;79:387-393. doi:10.1007/s10493-01900444-7
35. Snedecor GW, Cochran WG. Statistical methods. 8th ed. Ames: Iowa State University Press; 1994.
36. Solanki JB, Hasnani JJ, Patel DM, Patel PV, Raval SK. Canine demodicosis in Anand. J Vet Parasitol. 2007;21(1):79-80.
37. Sood NK, Mekkib B, Singla LD, Gupta K. Cytopathology of parasitic dermatitis in dogs. J Parasit Dis. 2012;36(1):73-77. doi:10.1007/s12639-011-0083-y
38. Sousa RFV, Gasparetto ND, Almeida BPF. Clinical and immunopathology aspects of canine demodicosis. IntechOpen; 2020. Available from: http://dx.doi.org/10.5772/intechopen.88763
39. Soulsby EJL. Helminths, arthropods and protozoa of domesticated animals. 7th ed. London: Baillière Tindall; 1982.
40. Suartha IN, Sudimartini LM, Wardhita AAGJ. Comparison of scraping, trichogram, and taping techniques for diagnosis of demodicosis in dogs. Jurnal Veteriner. 2018;19(1):85-90.
41. Swe PM, Zakrzewski M, Kelly A, Krause L, Fischer K. Scabies mites alter the skin microbiome and promote growth of opportunistic pathogens in a porcine model. PLoS Negl Trop Dis. 2014;8(5):e2897. doi:10.1371/journal.pntd.0002897
42. Valenciano AC, Cowell RL, Smith A. Cowell and Tyler's diagnostic cytology and hematology of the dog and cat. 4th ed. St. Louis: Elsevier; 2014.
43. Zorbozan O, Gerçeker Türk B, Acar A, et al. Comparison of skin scraping and standard superficial skin biopsy in the laboratory diagnosis of scabies. Turkiye Parazitol Derg. 2020;44(3):164-167. doi:10.4274/tpd.galenos.2020.6828