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Clinical Management of Papillomatosis in Bovines.

Introduction

Cutaneous papillomatosis is a chronic proliferative disease caused by DNA virus belonging to family papillomaviridae. Twelve different genotypes or strains of bovine papilloma virus (BPV) have been identified. BPV-1 and BPV-2 are associated with fibropapillomas in cattle and shows affinity for epithelial and dermal tissue (Pangty et al., 2010 and Somvanshi, 2011). Cutaneous papillomatosis is more common in bovine than other animals (Ozsoy et al., 2011).

Cattle are the main source and natural reservoir of infection by virus however halters, ropes and instruments can also serve as potential and indirect source of infection. Other factors that play a significant role in occurrence of disease are contaminated milking machines, malnutrition, hormonal imbalance and long term exposure to sunlight if there is immunodeficiency (Araibi et al., 2004). It can be transmitted from inapparent carrier to susceptible calf. Calves (young animals) are more susceptible and easily infected with papilloma virus through cut or abraded skin. However, takes 1-12 months to manifest as papilloma (Tan et al., 2012). The immunosuppressive factor plays an important role in progression of bovine papillomatosis as stated by Radostitis et al, (2007).

The incidence of cutaneous papillomas is reported to be much higher in crossbred than indigenous cattle (Eisa et al., 2000). Considering the above mentioned economic losses suffered by farmers, the present study was conducted with the objective to describe the incidence, clinical, haematological and histopathological aspects of bovine papillomatosis as well as to evaluate the efficacy of different treatments in cattle and buffaloes.

Materials and Methods

A total number of 1496 cattle and buffaloes of either sex of different age groups were screened for presence of cutaneous papilloma from December' 2012 to June' 2013 and 18 cases were included for the study.

The affected animals were clinically examined and data regarding location of lesion/papilloma, duration of illness (brief history), visual examination of lesion (i.e. ulcerative wound etc if any) were recorded. The haematological examination Two ml blood were carried out on 0th, 7th, 14th, 21st and 28th day. Gross lesions were recorded and representative tissue samples of growth were collected for histopathological examination. The cases were followed for at least three months for recurrence, if any.

The DNA from papillomatous tissue samples was isolated as per method of Sambrook et al. (1989). These extracts were tested by polymerase chain reaction (PCR) using primers specific to BPV1 (forward: 5'-ggagcgcctgctaactatagga-3' and reverse: 5'-atctgttgtttgggtggtgac-3'), BPV2 (forward: 5'-gttataccacccaaagaagaccct-3' and reverse: 5'-ctggttgcaacagctctctttctc-3'). The expected product sizes were 301 and 165 bp for BPV-1 and BPV-2, respectively. Cycling conditions were as follows:1 cycle of 94[degrees]C for 3 min followed by 30 cycles of 94[degrees]C for 40 sec, 52[degrees]C for 40 sec, 72[degrees]C for 1 min and final extension of 72[degrees]C for 10 min. The amplicons were visualized by 1 percent ethidium-bromide-stained agarose gel under UV transilluminator.

Results and Discussion

A total of eighteen (18) animals affected with papillomas were included in the present study, out of the 1496 screened animals. The average age of papilloma affected animals ranged from 2 months to 10 years. The papillomas grossly appeared in form of rough nodules and cauliflower like growth with horny papillae or dome shaped with a smooth outer surface (0.2-3 cm in diameter). The number varied from one to seventeen and size varying from pea nut to orange. The animals had single or multiple, hard, pedunculated or non-pedunculated horny warts which were present on head, neck, thorax, teats/udder, back and limbs. However, warts over the teats were of filiform like pinkish projections from the surfaces.

The highest incidence of 61.11 percent was recorded amongst young calves below 1 year of age. The higher incidence of papilloma in young animals could be due to undeveloped immune system. Ozsoy et al. (2011) also reported higher incidence of papilloma in young animals. The females (72.22 percent) were affected more than males (22.77 percent). The clinical examination of animals revealed that maximum number of papillomas/warts were on head region (44.44 percent) followed by thorax region (22.22 percent), neck region (16.66 percent), leg (5.55 percent), teat and udder (5.55 percent) and generalized papilloma (5.55 percent). Similar observations on incidence were recorded by Atasever et al. (2005) and Salib and Farghali (2011).

In present study, 919 cattle and 577 buffaloes were screened and higher incidence was observed in cattle (1.6%) than buffaloes (0.51%) which was in accordance with Sharma et al. (2004).

The non significant changes were recorded in Hb, TEC, TLC, PCV and DLC during haematological examinations in all animals suffering with cutaneous papilloma at the scheduled intervals.

The histopathological study of the section of cutaneous papilloma revealed varying degrees of hyperplasia of epidermis with irregular finger like papillary projections into dermis as a common texture (Fig. 6-8). The growths were categorized as fibropapilloma. Salib and Farghali (2011) reported similar papillate epidermal hyperplasia with patchy area of erosions, ulceration and neutrophil proliferation.

In epidermis, moderate to severe acanthosis, mild to severe hyperkeratosis, hydropic degeneration of keratinocytes and many koilocytes with variably sized keratohyalin granules were observed. Few cases revealed deposition of melanin pigment in large quantity (Fig. 9). Rare presence of intranuclear inclusion bodies was observed only in basal cells of epidermis. Dermis showed mild to moderate hyperplasia of connective tissue that consisted of blood vessels, fibroblasts, focal hemorrhage and mild infiltration of lymphocytes. The presence of mitotic figure in tissue was meager. Ozsoy et al. (2011) observed various degrees of acanthosis and hyperkeratosis.

Polymerase Chain Reaction (PCR) was done to detect BPV-1 and BPV-2 viruses in wart tissues of eighteen animals (Fig. 10). 165 bp amplified product corresponding to expected L-2 gene fragment of BPV-2 segment was observed in five samples out of eighteen samples. However, there was no amplification of BPV-1 segments from DNA of all eighteen samples. Singh and Somvanshi (2010) have also recorded presence of BPV-2 segment in papillomatous growth. Therefore it could be concluded that BPV-2 strain of bovine cutaneous papilloma was found prevalent in and around Nagpur region.

Assessment of Surgico-Therapeutic Regimen Ligation with Nylon - A total of six cases with small to medium sized pedunculated or stalked papillomas were tied using suitable ligature like nylon around the base tightly (Fig.1). In all cases, the papillomas started regressing in two week after being tied with nylon and sloughed off in 33.33% and 50.00% cases in five and seven weeks (Fig. 5), respectively. However in one case, the papilloma didn't slough off even after twelve weeks. It was then removed surgically after the observation period.

Treatment with Anthiomaline and Thuja (200x)

Total of six animals suffering with moderate to severe papillomatosis or generalized papillomatosis (Fig. 2) were treated with Anthiomaline @ 10-15 ml intramuscular at 48 hours interval for 3 consecutive doses and homeopathic drug Thuja - 200x @ 10-15 globule per day orally for thirty days. The extended doses of Anthiomaline were again administered in non responding cases. The papillomas started regressing after third dose of Anthiomaline and Thuja in all animals (Fig. 3). In 66.66% animals, papillomas regressed completely after 5th dose, however in 16.66% cases the complete regression was observed after completion of 6thdose on 20th day. In this case, Thuja was continued up to 20th day. In 16.66% case of papillomas on teat and udder, only 50% regression was observed even after 6th dose of Anthiomaline and Thuja. This case was further treated with same therapy for three more consecutive doses. The case responded positively and complete recovery after 25th day of administration was observed. Similar observations were recorded by Wadhwa et al. (1992) in cattle with same dose rate of Anthiomaline and Kumar et al. (1984) with Thuja.

Surgical Excision of Cutaneous Papilloma

Total of six animals were subjected to surgical intervention for removal of moderate to large sized or large solitary cutaneous papilloma in cattle and buffalo. The routine pre-operative preparation, pre-medication and anaesthetic protocol (Lignocaine hydrochloride 2%) was followed for all cattle and buffaloes subjected to surgical therapy. The operative procedure was completed within 15-25 minutes with minimum bleeding in five cases. However in one case, bleeding had to be controlled using electrocautery. Postoperative healing occurred uneventfully within 10-12 days in all animals (Fig. 4) and no recurrence was noted in follow up period.

Therefore from the observations of present study it could be concluded that all three treatment regimen were found have effect and has specific utility for papillomatosis in bovine. The small sized pedunculated or stalked papillomas could be well treated by tight ligation with nylon at the base, however, moderate to severe papillomatosis or generalized papillomatosis can be treated by combination of Anthiomaline and Thuja, whereas; the moderate to large sized or large solitary cutaneous papilloma of various location could be removed successfully by surgical excision.

References

Araibi, E.H., Marchetti, B., Ashrafi, G.H. and Campo, M.S. (2004). Down regulation of major histocompatibility complex class I in bovine papillomas. J. Gen. Virol. 85: 2809-14.

Atasever, A., Cam, Y. and Atalay, O. (2005). Skin papillomatosis in a cattle herd. Ankara Univ. Vet. Fak. Derg. 52: 197-200.

Eisa, M.I., Kandeel, A., El-Sawalhy, A.A. and El-Fetouh, M.S.A. (2000). Some studies on bovine papilloma virus infection in cattle with trials of its treatment. Vet. Med. J. Giza. 48: 47-55.

Kumar, A., Tanwar, R.K., Yadav, J.S. and Sharma, S.N. (1984). Treatment of bovine papillomatosis with some homeopathic drugs. Ind. J. Vet. Med. 4: 87-89.

Ozsoy, S.Y., Ozyildiz, Z. and Guzel, M. (2011). Clinical, pathological and immunohistochemical findings of bovine cutaneous papillomatosis. Ankara Univ. Vet. Fak Derg. 58: 161-65.

Pangty, K., Singh, S., Goswami, R., Saikumar, G. and Somvanshi, R. (2010). Detection of BPV-1 and -2 and quantification of BPV-1 by real-time PCR in cutaneous warts in cattle and buffaloes. Transbound Emerg. Dis. 57: 185-96.

Radostitis, O.M., Gay, C.C., Hincheliff, K.W. and Constable, P.D. (2007). Veterinary Medicine - A Textbook of the diseases of Cattle, Horse, Ship, Pig and Goats. Saunders Elseviers Publ. Newyork, USA. p. 1423.

Salib, F.A. and Farghali, H.A. (2011). Clinical, epidemiological and therapeutic studies on bovine papillomatosis in Northern Oases, Egypt in 2008. Vet. World 4: 53-59.

Sambrook, J. and Russell, D.W. (2001). Molecular cloning - A laboratory manual. Cold spring, Harbor Laboratory Press, New York. www.gobook.net/sambrook and russell/DNA isolation.

Sharma, S., K.B. Singh and B.K. Bansal (2004). Observation on teat papillomas in bovines. Ind. Vet. J. 81: 20-21.

Singh, V. and R. Somvanshi (2010). BPV-2 associated papillomatosis in Indian water buffaloes. Ind. J. Ani. Sci. 80: 956-60.

Somvanshi, R. (2011). Papillomatosis in buffaloes: a less-known disease. Trans. Emerg. Dis., Blackwell Verlag GmbH Publ. 58: 327-32.

Tan, M.T., Yildirim, Y., Sozmen, M., Dagalp, S.B., Yilmaz, V., Kirmizigul, A.H. and Gokce. E. (2012). A histopathological, immunohistochemical and molecular study of cutaneous bovine papillomatosis. Kafkas Univ. Vet. Fak. Derg. 18: 739-44.

Wadhwa, D.R., Rao, V.N., Mandial, R.K. and Prasad, B. (1992). Efficacy of anthiomaline and autogenous wart vaccine in bovine cutaneous papillomatosis. Ind. J. Vet. Med. 12 : 21.

Vijay Tailor (1), S.B. Akhare, M.G. Thorat, B.M. Gahlod and S.P. Salvekar

Department of Surgery and Radiology

Nagpur Veterinary College

Maharashtra Animal and Fishery Sciences University (MAFSU)

Seminary Hills

Nagpur - 440006 (Maharashtra)

(1.) Corresponding author. E-mail: sandeepakhare@yahoo.co.in
Table - 1: Age, sex, location of papillomatosis in bovines

                                  Number     Percent

Age        0-6 months               6        33.33
           6-12 months              5        27.77
           1-2 years                0         0.00
           2-4 years                4        22.22
           5 and above              3        16.66
Sex        Female                  13        72.22
           Male                     5        27.77
Location   Head                     8        44.44
           Thorax                   4        22.22
           Neck                     3        16.66
           Limb                     1         5.55
           Teat and udder           1         5.55
           Generalized papilloma    1         5.55
Species    Cattle                  15        83.33
           Buffalo                  3        16.67
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Title Annotation:Clinical Article
Author:Tailor, Vijay; Akhare, S.B.; Thorat, M.G.; Gahlod, B.M.; Salvekar, S.P.
Publication:Intas Polivet
Article Type:Report
Date:Jul 1, 2017
Words:1986
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