A Comparative pharmacological study on moxidectin and propolis ointment in rabbits naturally infested with Psoroptes cuniculi

A Comparative pharmacological study on moxidectin and propolis ointment in rabbits naturally infested with Psoroptes cuniculi

M.F. Eldakroury¹ and A.A. Darwish²

¹Department of Pharmacology, Faculty of Veterinary Medicine, Matrouh University, ² Animal and Poultry Health Department, Desert Research Center, Cairo, Egypt

[email protected], 0000-0001-9383-1433

[email protected], 0000-0002-8675-6313

2020-08-25

2021-01-01

Abstract

Ear mange is an annoying problem for all animal breeders including rabbit's breeders. This study aimed to compare the efficacy of moxidectin and 10% propolis ointment on rabbits infested with Psoroptes cuniculi with especial reference to some related biochemical parameters diagnostic and prognostic value. For this purpose, a total of thirty-two rabbits: 8 non-infested rabbits (control group (CG) and 24 naturally infested rabbits were used. Infested rabbits were equally divided into three groups: 1^st group remained without treatment (diseased group (DG)), 2^nd group was treated with moxidectin ((0.2 mg/kg body weight) S/C (MG)). Propolis ointment (10%) was applied topically to rabbits of the 3^rd group (PG) once daily. Ear scrapes and blood samples were taken on 0, 7, 14 and 21 days. The ear scraping was examined microscopically and the biochemical parameters were estimated and statistically analyzed. The results cleared that moxidectin achieved the complete healing (14^th day) faster than propolis ointment (21^st day), but MG suffered from a significant (P<0.05) hypoalbuminemia, increased liver and kidney function tests and cortisol levels when compared to PG. Liver and kidney function tests and cortisol yielded good values of the area under the curve, sensitivity%, specificity%, likelihood ratio, PPV%, NPV%, accuracy rate% in DG, MG (except creatinine) and PG. The study concluded that propolis had a prominent antipsoroptic effect without inducing hepatic or renal toxicity. Liver and kidney function tests and cortisol may be useful markers for rabbit mange diagnosis and prognosis and following up its treatment.

Keywords: Moxidectin, Propolis, Ear mange, Rabbits, Biochemical parameters

دراسة دوائیة مقارنة على عقار موکسیدیکتین ومرهم البروبولیس (صمغ النحل) فی الأرانب المصابة طبیعیاً بجرب الأذن

محمد فهمی الدکرورى¹ و أسماء عبدالله رجب درویش²

¹ فرع الأدویة، کلیة الطب البیطری، جامعة مطروح، ² فرع صحة الحیوان والدواجن، مرکز بحوث الصحراء، القاهرة، مصر

الخلاصة

یعد جرب الاذن مشکلة مزعجة لکل مربى الحیوانات بما فیهم مربى الأرانب. هدفت هذة الدراسة الى المقارنة بین کفاءة کل من عقار موکسیدیکتین ومرهم البروبولیس (صمغ النحل) فی القضاء على جرب الأذن فى الأرانب المصابة طبیعیا مع اشارة خاصة إلى القیمة التشخیصیة والتنبؤیة لبعض المتغیرات البیوکیمیائیة المرتبطة بالمرض. لهذا الغرض تم استخدام 32 أرنب: 8 أرانب سلیمة ظاهریا وبدون أی إصابات جلدیة (مجموعة ضابطة) و 24 أرنب مصابین بجرب الأذن. قسمت الأرانب المصابة الى ثلاث مجموعات: الاولى لم تتلقى اى علاج (المجموعة المریضة) والثانیة تم علاجها بعقار موکسیدیکتین (0.2 ملل/کجم تحت الجلد) والثالثة تم علاجها باستخدام مرهم البروبولیس موضوعیا بصورة یومیة. تم أخذ مسحات من الأذن وعینات دم فی الأیام 0، 7، 14، 21. تم فحص المسحات بالمیکرسکوب وتقدیر المتغیرات البیوکیمیائیة وتحلیلها إحصائیا. أوضحت النتائج قدرة عقار موکسیدیکتین على تحقیق الشفاء التام (الیوم 14) أسرع من مرهم البروبولیس (الیوم 21). ولکن عانت مجموعة الموکسیدیکتین من نقص معنویا فى ترکیز الزلال وزیادة معنویة فی وظائف الکبد والکلى بالمقارنة بمجموعة مرهم البروبولیس. کما حققت وظائف الکبد والکلى والکروتیزول قیم جیدة من المساحة تحت المنحنى، الحساسیة، المعنویة، نسب الاحتمالیة، القیم التنبؤیة الموجبة والسالبة ومعدلات الدقة فی المجموعة المریضة ومجموعة الموکسیدیکتین (ماعدا الکریاتینین) ومجموعة البروبولیس. هذا وخلصت الدراسة إلى أن مرهم البروبولیس له تاثیر فعال فى القضاء على الجرب بدون احداث تسمم فى الکبد او الکلى. وظائف الکبد والکلى والکروتیزول کواشف جیدة یمکن أن تساعد فی تشخیص الجرب والتنبؤ باستجابة الحیوان لخطط علاجه.

Introduction

 Psoroptes cuniculi is the main cause of rabbit's ear mange. It is a worldwide distributed and extensively threats caged rabbits breeding. Although, the disease first stages are non-fatal, but lowered body weights, fertility rates, treatment and labor cost associated with the disease and its control are economic losses can't be neglected (1,2). In addition, the disease complications may be lethal at chronic untreated cases. Moreover, the disease has a zoonotic importance. The major clinical signs of ear mange infestation include the presence of waxy material in external ear mixed with crusts, pain, pruritus, ulcer, hemorrhages, head shaking, scratching, and loss of body weight (2,3). Ivermectins are the first choice for rabbit mange treatment. Among them, moxidectin which is a second-generation macrocyclic lactone. It is a parasiticide active against a wide range of internal and external parasites (4). Unfortunately, macrocyclic lactone is not currently recommended for use in rabbits in many parts of the world as resistance (5), toxicity and environmental contamination and persistence(6). Interest is developing to use biocompatible products for external parasites in rabbits(7). Propolis is a natural resinous mixture produced by honeybees. Propolis means in Greek, entrance to the community this refers to its role in hive defense. Another name of propolis is bee glue due to its waxy nature and mechanical properties, bees use it for building or repairing hives and for sealing openings and cracks and smoothing out the internal walls (8). It was used in medicine by Greek and Roman for its antipyretic, antiseptic and wound healing propriety (9). It is a natural remedy for burns, sore throat, stomach ulcer, upper respiratory tract infections, common cold, and dermatological preparations for wound healing (10). Furthermore, it is effective in the treatment of dermatophytosis (11) and sarcoptic mange (6).

This work was conducted to study using 10% propolis ointment as an alternate to moxidectin in ear mange treatment with especial reference to the value of proteingram, liver and kidney function tests and cortisol in rabbit's ear mange diagnosis and prognosis and monitoring of the suggested therapeutic programs.

 Materials and methods

This study was carried out according to the ethical approval of Animal and Poultry Health Department, Desert Research Center (DRC), Cairo, Egypt.

Rabbits

Twenty-four Rabbits (average body weight 2.45 - 2.90 kg) had a history of anorexia, scratching, shaking their heads and ear and presence of crusty material in the ear canal. Microscopical examination of the lesions revealed that they were infested by with Psoroptes cuniculi. These rabbits had not been previously treated with any anti-acariasis drug. Eight non-infested rabbits (normal appetite, clean ear canal, no signs of irritation or restlessness), were used as a control group (CG). The other infested rabbits were divided into three equal groups each of 8 rabbits (according to body weight- extend of infection). The 1^st one (DG) was used as diseased group, group 2 (MG) received a single dose of moxidectin (0.2 mg/kg body weight) S/C. Propolis was applied topically to the rabbits of group 3 (PG) once daily. They were kept in individual cages, fed a balanced pelleted ration and clean source of water and libitum during the experimental period. All measures were performed to avoid re-infestation from the environment, rabbits were kept under observation (for clinical signs of mange infestation and medication toxicity). Ear scrapings were taken and examined microscopically under low power magnification according to Ulutas et al.(12) on 0^th, 7^th, 14^th and 21^st day.

Drugs

CYDECTIN^® (Contains 10 mg moxidectin/ml) is a product of Boehringer Ingelheim. 10% Propolis ointment was prepared according to Sanchez et al. (13) with some modifications.

Sampling

Blood samples were collected from ear vein of all rabbits at 0^th, 7^th, 14^th and 21^st day in clean sterilized centrifuge tubes without anticoagulant. The blood allowed to coagulate and the serum was collected and preserved at -20 ºC for determination of some biochemical parameters. Total protein (TP), albumin (Alb), liver enzymatic activity (aspartate aminotransferase (AST), Alanine aminotransferase (ALT)), kidney function tests (urea, creatinine (Cr)) were determined spectrophotometrically by using Biodiagnostic company^® commercial kits, Cairo, Egypt. While, serum cortisol levels were evaluated using commercial, coated tube radioimmunoassay kits, Pantex, Santa Monica, CA. Serum globulin (Glob) concentrations were obtained by subtracting the value of albumin from the corresponding value of total protein. A/G ratio by division of albumin on the corresponding globulin value.

Statistical analysis

SPSS program version 24 was used to compare between means of different statistical parameters (one-way ANOVA test), estimate the post-hoc differences between means (a multiple comparison Tukey’s HSD test) and determine correlations between the selected parameters (Pearson's simple correlation test). A difference was considerable significant at P

Results

According to the obtained data, the clinical signs intense increased along the experiment in DG. At 21^st day, the lesions became more prominent and diffuse and the microscopical examination showed a larger number of living mites (P. Cuniculi) in DG. In accordance, the biochemical parameters of DG (compared to CG) displayeda significant increase in total protein and globulin concentrations at 0^th day then TP significantly decreased at 7^th, 14^th, 21^st days, while Glob levels non-significantly changed at 7^th day and significantly decreased at 14^th, 21^st days. On the other hand, a significant decline was observed in albumin concentrations at 0^th, 7^th, 14^th, 21^st days. A/G ratio significantly reduced at 0^th, 7^th days then significantly elevated at 14^th, 21^st days. A significant increase was also detected in the liver enzymatic activity (AST, ALT), kidney function tests (urea, Cr) and cortisol concentrations at 0^th, 7^th, 14^th, 21^st days (Figure 1,2).

On contrary, a marked gradual improvement of mange clinical signs and a prominent regression of lesions intense, were noticed in both treated groups (MG and PG) from 7^th day post-treatment till the end of the experiment. Microscopically, a gradual decline of living mites count was observed in MG and PG compared to 0^th day. By 21^st day, all treated rabbits recovered completely and clinical signs disappeared and ear scrapes were clean (no living mites). In turn, the above mentioned parameters levels noticeably increased or decreased in MG and PG till approaching the CG levels (non-significantly changed with CG). TP returned to CG levels in MG and PG at 7^th day, Alb in PG at 21^st day, A/G ratio in MG at 14^th day and in PG at 21^st day, Glob in MG at 14^th day. While, MG recorded a significant decrease in Alb at 7^th, 14^th, 21^st days and in TP concentrations at 14^th day (in relation to CG). Glob levels significantly increased in PG compared to CG at 7^th, 14^th, 21^st days (Figure 1). Liver and kidney function tests and cortisol concentrations decreased towards CG levels, they non-significantly changed with CG at 7^th day for ALT, AST, Cr, cortisol in PG, at 14^th day for urea in PG and ALT, urea in MG, and at 21^st for AST, cortisol in MG but Cr significantly increased till the end of the experiment in MG compared to CG (Figure 2).

The comparison between MG and PG exhibited that: the lesions regression was faster in rabbits treated with moxidectin at 14^th day, then rabbits treated with10% Propolis ointment at 21^st day. While, a significant reduction in Alb at 7^th, 14^th, 21^st day and TP at 14^th day was depicted in MG. MG also showed a significant increase in AST at 14^th day, in ALT, urea, cortisol at 7^th, 14^th day, in Cr at 7^th, 14^th, 21^st day. In contrast, Glob significantly elevated in PG at 14^th, 21^st days (Figures 1,2).

In connection to the estimated biochemical parameters importance in the disease diagnosis and monitoring the diseased rabbits response to the two studied proposals, table 1 clarified that AST, ALT, urea, Cr and cortisol yielded high values of AUC in DG, MG (except Cr) and PG with acceptable values of sensitivity%, specificity%, LR (moderate to high), PPV%, NPV% and accuracy rate% along the experiment. In addition, they showed a significant positive correlation with the numbers of rabbits had clinical signs of psoroptes mange infestation (Table 2).

Figure 1: Number of rabbits showing clinical signs and serum levels of protein profile in the studied groups (N=8). Columns bearing different letters for the same day, differ significantly (P<0.05). CG: control group, DG: diseased group, MG: moxidectin group, PG: propolis group.

Figure 2: serum levels of liver and kidney functions tests and cortisol in the studied groups (N=8). Columns bearing different letters for the same day, differ significantly (P<0.05). CG: control group, DG: diseased group, MG: moxidectin group, PG: propolis group.

Table 1: Area under the curve (AUC), cut off points, sensitivity %, specificity %, Likelihood Ratios (LR), PPV%, NPV%, accuracy rate% in DG, MG and PG