Veterinary Research Forum
Volume:13 Issue: 2, Spring 2022

Due to the high prevalence of multi-drug resistant bacteria, combination therapy is an efficient choice for treatment of infections caused by highly resistant strains. In this study, the efficacy of ceftiofur plus danofloxacin combination was investigated against resistant Escherichia coli. The interaction between the two drugs was determined by checkerboard tests and time-kill assays. The combination was defined as bactericidal or bacteriostatic based on the minimum bactericidal concentration test results. Mutant prevention concentration test was used to evaluate the resistance tendency suppression potential of the combination. The combination had a synergistic effect against 83.00% of the isolates as verified by the checkerboard and time-kill assays. The combination was defined as bactericidal against all E. coli strains, since minimum bactericidal concentration: minimum inhibitory concentration ratios were below four thresholds and also markedly reduced mutant prevention concentration values of ceftiofur up to 4000-fold compared to its single use. Ceftiofur plus danofloxacin combination inhibited growth of E. coli strains which were resistant to ceftiofur or newer generation of fluoroquinolones. Our results suggest that ceftiofur plus danofloxacin combination has a bactericidal characteristic and can be an important alternative for the treatment of infections caused by resistant E. coli.

Lamb enteritis constitutes an economic burden on sheep production worldwide. We aimed to estimate the prevalence of Shiga toxin-producing Escherichia coli (STEC) and Salmonellae among diarrheic lambs at Kafrelsheikh Governorate, Egypt and to detect the associated clinical, hematologic, biochemical, and antioxidant parameters. Fifty diarrheic and twenty apparently healthy control lambs were examined clinically, and hematologically. Diarrheic lambs had a significant elevated body temperature, respiratory and pulse rate, most of hemogram para-meters, total proteins and albumin, oxidative stress markers malonaldiahyde and nitric oxide levels, liver enzymes, urea and creatinine than control group. On the other hand, these diarrheic lambs had significant reduction in total leukocyte count and lymphocytes, antioxidant biomarkers super oxide dismutase activities and reduced glutathione than control lambs. E. coli and Salmonella spp. were isolated from 32.00% and 16.00% of diseased lambs, respectively. Serotyping and biochemical tests of examined samples identified 16 E. coli isolates belonged to 10 different serotypes; O6, O8, O26:H11, O75, O84:H21, O103:H2, O114:H4, O121:H7, O128:H2 and O163:H2. All isolates are STEC as they harbor either Shiga-toxin 1 or Shiga-toxin 2 genes or both. One isolate carries intimin gene (eaeA) and classified as EHEC; O26:H11. The obtained nine isolates of Salmonella carry enterotoxin (Stn) genes, eight of them carry hyper-invasive locus (hilA) gene, all isolates belonged to six serotypes; S. Enteritidis, S. Heidelberg, S. Tsevie, S. Typhimurium, S. Essen, and S. Infantis. Lamb diarrhea was prevalent in the studied area and might constitute a veterinary and public health threat. Alteration in hemato-biochemical para-meters and oxidative–anti-oxidant balance could help adopt appropriate treatment regimens.

In order to prepare reductive polypeptides from the placenta of dairy cows，fresh placentas from healthy Chinese Holstein cows were obtained and homogenized. Response surface model was established to optimize the hydrolysis condition for the extraction of the placental polypeptides. Specifically, the placental tissue homogenate was treated with both trypsin and pepsin for 348 min and 329 min; at 35.00% and 35.75% of substrate concentration; with anenzyme-substrate ratio of 3.33% and 3.92%, respectively, based on the models. The treated samples were then demineralized and freeze-dried to obtain the hydrolyzed polypeptides. In order to identify the molecular mass distribution and reducibility of polypeptides, matrix-assisted laser desorption ionization (MALDI) and Prussian blue methods were used. The concentrations of placental polypeptides after hydrolysis by trypsin or pepsin were 5.52% and 5.97%, respectively; the vitamin C (Vit C) equivalents were 36.26 μg mg-1 or 61.15 μg mg-1, respectively. Both groups showed intensity peaks of MALDI patterns in the range of 300 - 400 Da, and polypeptides hydrolyzed by pepsin had higher Vit C equivalent anti-oxidant activity than trypsin hydrolyzed polypeptide, suggesting that the proteins in the placental tissues were hydrolyzed to di-peptides and tri-peptides completely. In conclusion, both trypsin and pepsin hydrolysis performed well in preparation of reductive polypeptides from the fresh placentas of dairy cows; while, pepsin is more effective than trypsin. The primary reductive ingredients may be the oligopeptides with molecular mass less than 1000 Da.

Piglets suffer from diarrhea caused by the Shiga toxin-producing Escherichia coli (STEC) and can be carriers of the bacteria, with public health consequences in developing countries. The aim of the present study was to study the prevalence of STEC O157 in feces of 465 piglets and 54 food mixes from backyard systems, the antimicrobial susceptibility of STEC and the frequency of genes encoding extended-spectrum β-lactamases. The E. coli was isolated from 75.90 % of the evaluated feces. The STEC strains were identified in 33.11% of the sampled population and in 43.60% of the piglets carrying E. coli. Among STEC strains, the stx1 gene was the most frequent (22.30%). The rfbO157 gene was amplified in 47.40% of the STEC strains. High frequencies of STEC strains were not susceptible to ampicillin, carbenicillin and tetracycline. The blaTEM gene (52) was the most frequent among strains not susceptible to ampicillin. Class 1 integrons were the most frequent in those strains. Of the identified STEC strains, 48.70% were considered as multi-drug resistant and 1.90% were considered extensively drug resistant. In the supplied food, STEC O157 strains were identified in 25.00% of the STEC strains. We conclude that the piglets from backyard systems are carriers of STEC O157 strains not susceptible to common antibiotics, including penicillins and tetracyclines. In addition, supplied food is a source of this type of pathogenic bacteria. Through their direct contact with humans, the piglets and food represent a potential source of bacterial dissemination capable of producing gastrointestinal infections in humans.

The study was designed to explore anatomical and histological vasculature changes in the female genital system of camel and serological aspects that might be responsible to maintain thermoregulation. Twenty-four adult female camels were sampled during breeding (November-April) and non-breeding (May-October) season. Blood was collected for estrogen, progesterone and cortisol level estimation. Genital organs were sampled and described after slaughtering. Samples were taken from the ovarian artery (OA), vein (OV) and arterio-venous complex (AVC), for light and scanning electron microscopy. Sections were stained with Hematoxylin and Eosin. (H&E), Masson’s trichrome, Weigert’s elastic and toluidine blue. Temperature and relative humidity were used to calculate stress indicator. Stress indicator was higher in non-breeding season (NBS). Anatomical and histological vasculature (OA, OV, AVC) dynamics were significantly higher in breeding season (BS) especially diameter of left OA. Parameters of OA were positively associated with estrogen level. Collagen, elastic, smooth muscles and mast cells were recorded least in BS compared to NBS. Unique venous structure, intra-mural venules (IMV), was discovered in tunica intima of OA, seen positively and negatively associated with estrogen and cortisol level in BS, respectively. Scanned electron-micrograph exhibited penetration and wrapping of OA by small thinned-walled venules that may form IMV. The AVC was too tightly packed to differentiate due to the collapse of the wall. Hormonal, seasonal, stress indicator and vascular dynamic of female genital system are interlinked and IMV in association with OA and OV may be proposed as the site of counter-current exchange in female reproductive system of the camel.

This study aimed to evaluate the protective effect of cinnamic acid (CA) on orthophenyl-phenol (OPP)-induced oxidative stress in rats. Thirty-two Sprague-Dawley male rats were divided into four groups as control, OPP, CA and OPP + CA groups. The animals in control, OPP and CA groups were received corn oil, OPP (700 mg kg-1 dissolved in corn oil) and CA (200 mg kg-1) by gavage for 21 days, respectively. The animals in OPP + CA group were received CA for 3 days and from day 4; OPP and CA were applied together daily until day 25. Blood and liver samples were collected at the end of experiment for measurement of aminotransferases, creatinine (CREA), catalase (CAT), superoxide dismutase (SOD), glutathione peroxidase (GSH-Px), reduced glutathione (GSH) and malondialdehyde (MDA). The OPP-induced high serum activities of aminotransferases and level of CREA were reduced significantly by CA administration. The OPP induced significant increases of CAT activities and MDA levels in serum and liver tissue samples. Moreover, OPP significantly decreased GSH levels as well as GSH-Px and SOD activities. Pre-treatment with CA ameliorated the GSH levels along with GSH-Px and SOD activities compared to OPP-receiving rats. On the other hand, CAT activities and MDA levels significantly decreased following the pre-treatment with CA compared to OPP-receiving rats. It has been determined that OPP causes oxidative stress and lipid peroxidation in blood and liver tissues and creates changes in anti-oxidant defense enzymes. Pre-treatment with CA reduces lipid peroxidation and provides protective effect against oxidative stress.

Bone-marrow derived stem cells (BMSCs) can differentiate into several mesenchymal cell lines and are suitable candidates for bone and dental tissue engineering. This study aimed to assess the efficacy of cell therapy in direct pulp capping (DPC) of canine teeth using autologous BMSCs along with collagen/hydroxyapatite hybrid scaffold in terms of the quantity and quality of calcified bridge formation. The teeth were randomly divided into three groups of DPC with mineral trioxide aggregate (MTA), hydroxyapatite/collagen hybrid scaffold alone and BMSCs with hydroxyapatite/collagen hybrid scaffold; DPC was performed under general anesthesia in cavities prepared on the buccal surfaces of mandibular and maxillary premolars of the same dogs from which, stem cells had been isolated. All cavities were then restored with light-cure resin modified glass ionomer cement. Histomorphometric assessments after 12 weeks showed formation of dentinal bridge following DPC with BMSCs and MTA. The efficacy of MTA for calcified bridge formation following DPC was significantly higher than that of BMSCs plus hybrid scaffold (P<0.0001). According to present study, we concluded DPC using BMSCs and hybrid scaffold does not provide clinically noticeable results in canine patients.

Phthalates are environmental contaminants mostly used as plasticizers and additives in different products. Having endocrine-disrupting properties, phthalates are known as potential reproductive toxicants. The present study was conducted to evaluate the reproductive toxicity of di-n-butyl phthalate (DBP) in pregnant rats and their offspring and also to assess the ability of vitamin E in the elimination or reducing reproductive toxicity of DBP. Sixty-six pregnant Wistar rats were exposed to 100, 500 or 1,000 mg kg-1 per day DBP or 500 mg kg-1 per day DBP along with 100 mg kg-1 per day vitamin E during gestation. After delivery, they were divided into two groups. In one group gavage was finished after litter while in the other DBP administration was continued till weaning. The results showed that DBP affected many aspects of reproductive performance in pregnant rats and their offspring. It could be suggested that vitamin E could ameliorate the adverse effects of DBP, especially in male pups.

Limited knowledge is available on acute intoxication with environmental toxicants in birds. This experimental study determines features of acute caffeine intoxication and clinical outcome of different treatments in pigeons. The experiment was performed in three phases. Toxicity tests were performed in phases one and two while phase three was dedicated to comparative evaluation of fluid therapy and activated charcoal with or without diazepam and/or propranolol on clinical outcome of birds. Calculated LD50 was 366 mg kg-1 although presence of regurgitation compromised the accuracy of LD50 application. The dose-response (death) curve was sharp with a slope of 8.41. Clinical signs included renal, neurological, gastrointestinal and respiratory symptoms that generally initiated in a few minutes and lasted for few hours. The approximate toxic dose (ATD) was 294 mg kg-1. Serum and brain concentrations after administration of ATD followed a normal distribution in a range of 14.6 - 83.3 mg mL-1 and 1.04 - 7.81 µg g-1, respectively. Fluid therapy and activated charcoal with or without propranolol did not affect the clinical outcome of intoxicated birds while adding diazepam and intensive therapy with all of these agents even worsened the situation. In conclusion, caffeine is a potential source of intoxication in pigeons with a fast onset of clinical signs and a sharp increase in death rates by increasing doses. Symptoms are similar to mammals with prominent neurological signs although the ATD and serum concentrations are relatively high. Intensive therapy with above mentioned drugs is not recommended. Most birds survive after resolution of transient clinical signs without any special treatment.

The aim of this study was to introduce a new animal model of fecal incontinence (FI) by injecting abobotulinumtoxinA in the external anal sphincter (EAS) muscle of dogs which replaces models based on anal sphincter destructions that are invasive, mostly require surgical procedures, expensive, permanent, and painful to the animals. 4 healthy mongrel dogs were used in this study. First, they were received NaCl 0.09% (as control) injections in EAS muscle and effects were assessed by means of Electromyography (EMG) and clinically evaluated by sphincter pinch test and presence of leakage of feces for 2 weeks. Then, they received abobotulinumtoxinA in EAS muscle and reevaluated for 6 weeks to see short-term and medium-term effects of abobotulinumtoxinA injection. Saline had no significant changes in results obtained from EMG, however, there were significant decreases in amplitudes of action potentials after receiving abobotulinumtoxinA in comparison with no injection or saline injection in EAS muscle. Pinch tests were normal after saline injection assessment period, however, then started to be negative, ranging from two days after abobotulinumtoxinA injection to seven days after receiving abobotulinumtoxinA. Animals also had significant presentations of fecal incontinence (leakage of feces and cage contamination with feces) from the 1st week after receiving abobotulinumtoxinA until the 6th week after receiving abobotulinumtoxinA. AbobotulinumtoxinA caused paralysis in the EAS and producd FI conditions in dogs. This animal model was an appropriate substitute to the various invasive, expensive and also complicated procedures with an easy, feasible, noninvasive and non-painful single-stage abobotulinumtoxinA injection.

In addition to the fifty years since the test-and-slaughter program began in Iran and despite a significant reduction in the disease prevalence, positive tuberculosis cases are still being isolated from livestock farms across the country. Tests with 100% sensitivity and specificity are essential features for bovine tuberculosis diagnosis. The relationship between real-time PCR and immunohistochemistry (IHC) as two essential laboratory methods in the diagnosis of bacterial infections were aimed to evaluate single intradermal comparative tuberculin test (SICTT) results. One hundred thirty-eight cows in two groups were examined: Reactors (108 cows) and clean (as a control group; 30 cows). In the reactor group, 58(54.00%) cows were Mycobacterium bovis positive, 46(43.00%) were Mycobacterium avium subsp. paratuberculosis (MAP) positive, and 11(10.00%) were Mycobacterium tuberculosis positive. 32(55.00%) cows were co-infected with M. bovis and MAP and 5(4.55%) cows were co-infected with Mycobacterium tuberculosis and MAP in this group. Of 50 M. bovis negative cows of reactor group were 14(28.00%) MAP positive and 36(72.00%) negative, as well. Concurrent infection with all was observed in one reactor case. Comparing IHC and real-time PCR for the detection of bovine tuberculosis and Johne’s disease showed very good agreement (Kappa values 0.81 - 1.00). The results also provided further confirmation on IHC and real-time PCR as a sensitive and reliable diagnostic screening approach for detection of bovine tuberculosis. The use of one laboratory method to detect bovine tuberculosis is not sufficient alone.

Skin flap necrosis has been remained as an unsolved problem in plastic and reconstructive surgeries. Here, we explored the effects of metformin post-treatment on random skin flap survival in rats. An 8.00 × 2.00 cm dorsal skin flap was created in 24 rats and they were then divided into three groups (n = 8) including Control, metformin (Met) 50.00 mg kg-1 and Met 100 mg kg-1. All animals were administrated orally until seven days after flap surgery. Flap survival, the number of blood vessels and mast cells in the flap tissues were analyzed. Vascular endothelial growth factor (VEGF) expression levels in flap tissues was also determined using immunohistochemical methods. The percentage of survival area in Met 50.00 mg kg-1 and Met 100 mg kg-1 groups were significantly higher compared to control. The blood vessel density and the VEGF positive cells in the viable areas of flaps showed a significant increase in Met 50.00 mg kg-1 group compared to control group. The results of this study suggested that treatment with metformin, especially with low dose following skin flap surgery was effective in improving the flap survival and increasing the neovascularization in the flaps tissues of rats.

Some pharmacological agents can be effective for peripheral nerve injuries treatments. Present study aimed to apply different agents and compare the nerve regenerative effects following crushed sciatic nerve injuries. Twenty four (n=24) adult male mice were conducted in this study. Standard unilateral left side sciatic nerve crush was performed with 2 mm width mosquito hemostat forceps. The mice were randomly divided into 4 groups with the same numbers in each group which received subcutaneously, estrogen (group I), tacrolimus (group II), the combination of estrogen and tacrolimus (group III), and saline 0.9%. Functional recovery, histopathology, and Immunohistochemistry (IHC), were assessed on days 14th and 28th. Walking track analysis on day 14th showed no significant difference between experimental groups (P>0.05) but they showed significant difference compared to the control group (P<0.05). At the same time, experimental groups showed similar results of inflammatory cell infiltration, axonal edema, and count with significant differences between the control (P<0.05). At the end of the study, group I and III showed a significant difference in functional recovery between group II and control (P<0.05). After fourth week significant histopathological difference of axonal count was observed in group III (P<0.05). On day 28th, only IHC assessment in group III increased more Glial fibrillary acidic protein (GFAP) expression compared with the same group on day 14th. This study revealed subcutaneous administration of combined estrogen and tacrolimus can be effective with acceptable results in nerve regeneration.

Cyclophosphamide leaves an undesirable effect on testes. This study was conducted to explore the effects of the Phoenix dactylifera (DP) on testes following the cyclophosphamide treatment. Thirty-six male mice were divided into six groups, one control, one cyclophosphamide, two groups of cyclophosphamide with a dose of 200 mg kg-1 and a dose of 400 mg kg-1 DP, and two of only high and low doses of DP. All groups were gavaged daily for 28 days. The animals were euthanized 24 hr after implementing the last treatment. Then, the testes and epididymis samples were removed and weighed. The main sperm characteristics such as the number of sperm and sperm viability, the morphometric changes, biochemical analysis of testes and enzyme activity were investigated. With the cyclophosphamide group, only body weight, testes weight, epididymis weight, sperm viability and the fertilization percentage were decreased significantly compared to the control group. Moreover, the spermatogenesis indices and morphometric parameters in this group indicated a significant decrease. Furthermore, the morphological changes were observed in the testicular tissue, including seminiferous tubule atrophy, vacuolation, hyperemia of blood vessels and increased space in the interstitial tissue. In the biochemical study of cyclophosphamide group, the amount of glutathione peroxidase in serum was decreased, whereas, the amount of malondialdehyde in testicular tissue showed a significant increase. The DP group included the antioxidant and anti-apoptotic properties. It seemed that the compounds in the DP would lead to the inhibition of the production of active metabolites released from the cyclophosphamide.

The aim of this study was to evaluate the correlation between some fetal parameters measured by ultrasonography and fetal age for developing a formula which could be used for estimation of fetal age in Baluchi sheep. Placentome diameter (PD), gestation Sac diameter (GSD), biparietal diameter (BPD), straight crown-rump length (CRL), femural length (FL), occipito-snout diameter (OSD), thoracic diameter (TD) and fetal heart rate (FHR) were measured by transrectal and/or transabdominal ultrasonography. All ewes were synchronized using progesterone sponge for 6 days. On the first day, they received PGF2 and on the last day received 400 IU of equine serum gonadotrophin (eCG). All of the parameters revealed significant correlation by increasing fetal age. Except for PD, for other values linear regression curve was illustrated. BPD (between 36 - 96 days) (R2 = 0.961), CRL (R2 = 0.935), FL (R2 = 0.950), OSD (R2 = 0.981), TD (R2 = 0.975) showed high correlation with fetal age (p < 0.001). Moderate correlation was calculated for FHR (p < 0.001, R2 = 0.883). Low correlations were assessed by measurement of GSD (p = 0.018, R2 = 0.318), BPD (between 96-138 days; p = 0.038, R2 = 0.29) and PD represented the significant non-linear correlation with age (p < 0.001), maximum correlation was assessed by measurement of mean placentome wall diameter. In conclusion, OSD was recommended as the best parameter for estimating of gestational age between days 36 - 109 in Baluchi sheep because of the lowest residuals, the highest correlation coefficient and wide period of availability for imaging in gestation.

Bovine Leukemia Virus (BLV) is an oncogenic retrovirus of the genus Deltaretrovirus. The genome of BLV encodes a cluster of 10 mature microRNAs (miRNAs). Considering the importance of miRNAs in regulating gene expression, it seems that each of the miRNAs of BLV plays a vital role in the process of pathogenesis and tumorigenesis of the virus. First, sequences of each of the miRNAs of BLV were selected and downloaded from the miRBase database. The sequences were then investigated using TargetScan and miRWalk to identify target genes of each of the mature miRNAs of the virus. Second, the expression levels of the two miRNAs with the highest number of target genes in B lymphocytes and lymphoid tissues were evaluated using qPCR and were compared between cattle with different forms of BLV infection: PL form was compared to ‎aleukemic (AL) form (Group 1) and BLV+ with normal lymph nodes were compared to lymphosarcoma form (Group 2). We identified a total of 1595 target genes of the micro RNAs. The miRNAs with the highest target genes included miR-B4-3p with 760 and B2-5p with 102 target genes. In the second phase, miRNA expression in BLV-infected animals was investigated. The Fold Change (FC) values for miR-B4-3p and miR-B2-5p in group 1 were 22 and 67, respectively. In the second group, the FCs for miR-B4-3p and miR-B2-5p were 47 and 133, respectively. The expression was significantly higher in persistent lymphocytosis (PL) cattle in group one and lymphosarcoma cattle in group two.

Otitis externa is an inflammatory disease of the external ear canal; a therapeutic alternative is the controlled-flux electrolyzed acidic solution (SAEFC) with bactericidal and bacteriostatic properties. The purpose of this study was to evaluate the clinical and cytological efficacies of SAEFCin otitis externa. A total of 30 dogs were examined. The clinical and cytological evaluations were on days 1, 7 and 14; the following was considered: Pain, erythema, exudate, inflammation, ulcers, putrid odor and pigmentation. Cytological evaluations were carried out to examine the presence of polymorphonuclear leukocytes (PMN), intra-cellular bacteria (cocci; IC), extra-cellular bacteria (EC) and yeasts on days 1, 7 and 14. A cotton swab saturated with SAEFC was applied for each dog’s ear. The values of PMN diminished having a significant difference between days. The presence of IC (cocci) had a difference in evaluations from day 1 to day 7 and from day 7 to day 14 demonstrating an important drop. The evaluation of pain showed a sustained decrease until reaching 95.00% of remission, erythema diminished 68.33%, the exudate dropped to 90.00%, inflammation was no longer present by day 14 in 78.00% of the cases and 21.67% showed a slight inflammation, the putrid odor dropped to 95.00% and 5.00% presented a mild odor and by day 14 ear pigmentation was no longer present in 51.67% of the cases and the 48.33% had slight pigmentation. The use of SAEFC is an effective alternative treatment in dogs with otitis externa.

Anti-microbial resistant genes could be passed to human via the food chain or by direct contact with infected birds. To evaluate tetracycline resistance genes in the feces of companion birds suspected to enteritis, 100 fecal samples were collected from diarrheic companion birds in Isfahan province, Iran. The presence of Escherichia coli was examined by bacteriological, biochemical, and polymerase chain reaction (PCR) tests. The presence of genes associated with resistance to tetracycline (tetA, tetB, tetC, tetD, tetE, tetG, tetK, tetL, tetM, tetO and tetS genes) was examined using a multiplex PCR. The results showed that in enteric birds, 43.00% of fecal samples contained E. coli. In 26 resistant E. coli, 11, 12 and 3 strains contained tetA (42.30%), tetB (46.15) and tetA plus tetB (11.53%) resistant genes, respectively. In conclusion, E. coli isolates from the enteric problem of companion birds contained tetracycline resistant genes that may transfer to human and pose a risk for antibiotic effectiveness in the treatment of infectious diseases in human.

A 9-year-old spayed female Terrier dog was evaluated for lethargy, anorexia, polyuria, polydipsia and dysuria. The dog had been diagnosed with diabetes mellitus about 6 months ago and received subcutaneous doses of insulin. The patient showed insulin resistance and severe persistent fasting hyperglycemia in the face of high-dose insulin treatment, hyper-cholesterolemia and urinary tract infection. After a complete evaluation, the dog was diagnosed with a polycystic ovary and a cystic uterine remnant during an exploratory celiotomy. The polycystic ovary and cystic uterine remnant were removed and submitted for histopathological evaluation. Two weeks after surgery the blood glucose level and one month later serum cholesterol level were controlled, using a low level of insulin therapy (0.25 IU kg-1 Neutral Protamine Hagedorn (NPH) insulin, every 12 hr). In the present study, the clinical and laboratory results showed that ovarian remnant syndrome as an infrequently encountered condition in dog was related to some metabolic disorders such as insulin resistance, uncontrolled hyperglycemia, dyslipidemia and also recurrence urinary tract infection. To the best of authors’ knowledge, no reports of hypercholesterolemia in dog have been made before as a complication of ovarian remnant syndrome.

A 14-year-old female Shih Tzu was presented with conjunctivitis, keratitis, an enlarged mass located on the left lower eyelid and a large subcutaneous cyst in the temporal region. The hematological and biochemical analyses revealed no significant issues. The mass and the cyst were taken from the base with 1.50 cm of the underlying tissue. On histological examination, the mass was made up of adenoid arrangements of sebocytes and basaloid reserve cells with low to moderate mitotic activity of basaloid cells. In addition, a large number of meibomian ducts with marked infiltration of mononuclear inflammatory cells and melanin pigment were observed in the ductal part of the tumor. The cyst was lined by a simple cuboid-like epithelium with no eosinophilic cuticular lining. Immuno-histochemical staining of the mass with CK14 showed positive reaction in the neoplastic basaloid reserve cells and adipocytes; but, not with CK18 and vimentin. Unlike, the luminal cells of the cyst were stained with CK18. The co-existence of mixed meibomian gland and ductal adenoma with an apocrine sweat gland cyst was diagnosed based on the clinical, histological and immunohistochemical findings.