Deterioration of wound healing and intense suppression of MMP-9 mRNA expression after short-term administration of different topical glucocorticoids or NSAIDs in an avian model of corneal lesions.

BACKGROUND: Corneal lesions are considered among important ophthalmic conditions in avian patients. Short-term outcome of using anti-inflammatory agents in corneal lesions of birds are not well-described. AIMS: The study evaluates effects of different anti-inflammatory agents on healing of alkali burn-induced corneal lesions in layer hens as an avian model. METHODS: Adult layers were randomly allocated into 7 groups (n=15) as follows: 1. Negative (normal) control (NC), and 2. Positive control (PC) with an experimentally induced-corneal lesion, 3-7. Birds with corneal lesions that were treated with dexamethasone, fluorometholone, prednisolone, ketorolac, or diclofenac eye-drops every 6 hours (QID) for 5 consecutive days. RESULTS: At the end of the experiment, proper healing was observed in PC group based on lesion area, while treated groups showed statistically larger lesion sizes as compared to PC birds (P<0.05). Although no significant difference was observed among groups, birds treated with ketorolac, diclofenac or fluorometholone had higher histopathological scores for most of the assayed parameters than other groups. Levels of tumor necrosis factor-α (TNF-α) and vascular endothelial growth factor (VEGF) in corneal tissue of different groups were statistically the same. The mRNA expression of matrix metalloproteinase-9 (MMP-9) was increased 2.5 folds in PC group as compared to NC birds. However, birds treated with anti-inflammatory agents showed no detectable expression of MMP-9 mRNA. CONCLUSION: Five days of topical administration of non-steroidal anti-inflammatory agents (NSAIDs) or glucocorticoids (GCs) is associated with suppression of MMP-9 mRNA expression in corneal tissue and detrimental effects on wound healing in layers with alkali burn-induced corneal ulcers.

Morphological and molecular analyses of the spiruroid nematode, Falcaustra araxiana Massino, 1924 (= Spironoura araxiana) from the European pond turtle (Emys orbicularis).

There is little information on the phylogenetic position and life cycle of family Kathlaniidae. Falcaustra araxiana is a member of this family which infects the large intestine of the European pond turtle (Emys orbicularis). In the present study, morphological data and molecular analyses based on the 18S rDNA were performed on different types of F. araxiana originating from the large intestine and gastric nodules in the turtle. Morphological data revealed both larvae and adult stages in the gastric nodules. In addition, all nematodes recovered in the large intestine were adult worms. GenBank accession numbers KM200715 and KM200716 were provided for adult F. araxiana located in the intestine and stomach, respectively, of E. orbicularis. The results of sequencing proved that these two types are completely similar. Accordingly, it can be hypothesized that nodule formation is a part of the life cycle of the parasite or a survival strategy. Furthermore, F. araxiana develops to the adult stage in the gastric mucosa prior to migrating to the large intestine. Phylogenetic analysis revealed that F. araxiana unexpectedly branched away from other members of the superfamily Seuratoidea (Truttaedacnitis truttae, Cucullanus robustus and C. baylisi) and showed a closer relationship with Paraquimperia africana, a member of the Ascaridoidea. It seems that phylogenetic reconstruction for the present parasite needs more detailed morphology, life cycle and molecular studies.

Comparative evaluation of therapeutic efficacy of sulfadiazine-trimethoprim, oxytetracycline, enrofloxacin and florfenicol on Staphylococcus aureus-induced arthritis in broilers.

Staphylococcus aureus is an important human and veterinary pathogen that causes economic loss in the poultry industry. This study aimed to compare therapeutic efficacy of 4 commonly used antibiotics in poultry on S. aureus-induced arthritis in broilers. Sixty broilers, 8 weeks of age, were assigned at random into 7 groups as follows: (1) negative control (n = 5); (2) vehicle control (n = 5); (3) sulfadiazine-trimethoprim, 250 ml/1000 l drinking water (n = 10); (4) oxytetracycline 20%, 1 mg/l drinking water (n = 10); (5) florfenicol 10%, 1/1000 v/v in drinking water (n = 10); (6) enrofloxacin 10%, 1/1000 v/v in drinking water (n = 10) and (7) positive control (n = 10). Birds in group 2 were injected with 1 ml of sterile TSB medium into the right tibiotarsal joint on d 0 while other birds (except group 1) were challenged with 1 ml of 1.2 × 10(10) CFU/ml suspension of S. aureus bacteria. Antibiotic therapy was started from d 4 post challenge and continued for 5 d. At the end, birds were weighed and clinical severity of arthritis was determined. After blood collection, birds were slaughtered and tibiotarsal and hip joints were evaluated grossly. The content of inflammatory exudates of tibiotarsal joint and the degree of femoral head necrosis were recorded. Mucin clot test and histopathological evaluation were performed on right tibiotarsal joint. Serum interleukin 6 was also assayed. Sulfadiazine-trimethoprim had higher therapeutic efficiency with regard to most of the assayed criteria, whereas none of the antibiotics significantly affected femoral head necrosis and body weight. These data will help clinicians to have better antibiotic choice in field conditions.

Correlation between some hematological parameters, acute phase proteins and serum immunoglobulins in experimental caprine besnoitiosis.

The present study was undertaken to investigate correlation between some hematological parameters, acute phase proteins and immunoglobulins in the experimentally infected goats with Besnoitia caprae from the time of infection till 360 days post infection (DPI). Six male goats, approximately 12-16 months old, were inoculated subcutaneously with approximately 1.3 × 10(8) bradyzoites of B. caprae and blood samples were collected at weekly intervals from the jugular vein of the goats. Total leukocyte count and differential leukocyte counts were determined. Acute phase proteins (APPs) including serum amyloid A (SAA), haptoglobin (Hp), fibrinogen and ceruloplasmin were undertaken at weekly intervals. We evaluated an enzyme-linked immunosorbent assay (ELISA) (using a somatic antigen of bradyzoite) to detect anti-B. caprae antibodies in caprine sera. Cysts were present in the skin biopsies of the distal parts of the leg of the infected goats from 28 DPI. From 30 to 360 DPI, results showed that the APPs concentrations including SAA, Hp, fibrinogen and ceruloplasmin were enhanced in the serum of infected goats. However, there were some variation in hematological parameters; the differences were not significant with those of the normal values. Some variations were seen in the levels of specific antibodies against this parasite and they had correlation with some hematological parameters and acute-phase proteins.

First isolation and establishment of in vitro culture of Theileria lestoquardi from a naturally infected cow.

Theileria infected cell line was isolated from the prescapular lymph node of an adult crossbred cow. Molecular study confirmed this cell line of bovine lymphocyte has been transformed by the Theileria lestoquardi. This strain of T. lestoquardi designated Ka-6 and sheep were inoculated with this strain didn't show any clinical signs of theileriosis which shows the significance of this cell line to develop a tissue-culture vaccine against malignant ovine theileriosis. Contrary to accepted belief that the T. lestoquardi not capable of causing disease in cattle, the present study describes the first isolation and establishment of in vitro culture of T. lestoquardi-infected cell line from a naturally infected cow with typical singes of acute theileriosis.

Comparative assessment of the therapeutic effects of the topical and systemic forms of Hypericum perforatum extract on induced oral mucositis in golden hamsters.

Oral mucositis is a common and irritating complication of chemotherapy and radiotherapy for malignancies. Current treatments have failed to achieve complete remission of this complication. The St. John's wort plant (Hypericum perforatum) has long been known for its anti-inflammatory and antibacterial effects. The current study was designed to investigate the therapeutic efficacy of the topical and systemic administration of H. perforatum extract on oral mucositis. Oral mucositis was induced in 72 male golden hamsters by administration of 5-fluorouracil (60mg/kg), on days 0, 5, and 10 of the study. The cheek pouch was scratched with a sterile needle on days 1 and 2. On days 12-17, H. perforatum extract topical gel 10%, oral H. perforatum extract (300mg/kg), and gel base groups were treated and then compared with a control group. Weights and blood samples were evaluated, biopsies from buccal lesions were examined histopathologically, and tissue malondialdehyde (MDA) was measured. Both of the H. perforatum extract treatment groups saw a significant relief in oral mucositis compared to the control and base gel groups; the systemic form was superior to the topical form. H. perforatum extract, administered orally or topically, expedited the healing of chemotherapy-induced oral mucositis in hamsters.

Evaluation of Cross Immunity and Histopathological Findings in Experimentally Infected BALB/c Mice with Neospora caninum and Besnoitia caprae.

BACKGROUND: Caprine besnoitiosis is an economically important disease of goats. Neospora caninum, another coccidian parasite of worldwide distribution, infects several animal species and is a major cause of abortion in cattle. Combined infections of N. caninum and Besnoitia caprae can occur in geographical areas endemic for both species of parasite in goats. This experiment was conducted to investigate the possible cross-immunity between these two infections in experimentally infected BALB/c mice. METHODS: Forty BALB/c mice were divided into four equal groups. The mice of Groups 1 and 4 were inoculated with 1×10(6) live virulent tachyzoites of N. caninum (NC-1), while animals of Groups 2 and 3 were inoculated with sterile tissue culture medium. Each mouse in Groups 1 and 2 was challenged 28 days later with 1×10(6) live virulent bradyzoites of B. Caprae (BC-1). RESULTS: Following the challenge, the mice in Groups 1 and 2 showed 100% morbidity and 100% mortality within 9 days post infection, while all the animals of Groups 3 and 4 remained alive. The dead animals were necropsied. The survivors (mice in Group 3 and 4) were euthanized 9 days after inoculation and the gross and histopathological lesions in different organs were investigated. CONCLUSION: Immunization and challenge experiments with lethal dose of B. caprae in the highly susceptible BALB/c mice showed no cross-protection between N. caninum and B. caprae.

Histopathologic and ultrastructural studies on experimental caprine besnoitiosis.

The distribution pattern and associated tissue reactions with progressive changes in Besnoitia caprae cysts were investigated in 6 experimentally infected 16- to 20-month-old male goats. Each goat was subcutaneously inoculated with approximately 13 × 10(8) B caprae bradyzoites. The animals were examined daily for development of clinical besnoitiosis, and skin biopsies from distal parts of the limbs were taken at weekly intervals. At 15, 30, 60, 120, 180, and 365 days postinfection (DPI), 1 goat was euthanized. Samples were collected at autopsy from various organs for histologic and ultrastructural studies. No cysts were seen in tissue sections on 15, 30, and 365 DPI, but large numbers were present at 60, 120, and 180 DPI in the skin of the distal limbs, scrotum, and ears, with fewer in the tongue, palate, sclera, testicles, and spermatic cord. No cysts were seen in the lungs, liver, kidneys, spleen, central nervous system, or lymph nodes. Cyst numbers peaked at 60 DPI, then declined from 120 to 180 DPI. Degenerated cysts were relatively rare at 60 DPI but more numerous at 180 compared with 120. A granulomatous reaction--predominantly characterized by macrophages, lymphocytes, and plasma cells--surrounded each degenerated cyst. All goats showed testicular tubular degeneration with little or no spermatogenic activity. The sizes of cysts and their wall thickness, with the size of bradyzoites and some of their organelles, exhibited progressive chronologic changes.

Seroprevalence of Neospora spp. in horses in South of Iran.

Neospora caninum, an apicomplexan protozoan parasite, is recognized as a major cause of abortion in cattle. However, limited information is presently available on the seroprevalence of Neospora antibodies in horses worldwide. The aim of the present study is to determine serological prevalence of Neospora infection in horses in Iran. Blood samples were obtained from 200 horses and tested for serum antibodies against Neospora spp. by the Neospora modified direct agglutination test (N-MAT). Antibodies were found in 64 (32%) horses being tested with titers of 1:80. This is the first serological survey for Neospora antibodies performed on horses in Iran.

Seroprevalence of Neospora spp. in horses in South of Iran

Neospora caninum, an apicomplexan protozoan parasite, is recognized as a major cause of abortion in cattle. However, limited information is presently available on the seroprevalence of Neospora antibodies in horses worldwide. The aim of the present study is to determine serological prevalence of Neospora infection in horses in Iran. Blood samples were obtained from 200 horses and tested for serum antibodies against Neospora spp. by the Neospora modified direct agglutination test (N-MAT). Antibodies were found in 64 (32%) horses being tested with titers of 1:80. This is the first serological survey for Neospora antibodies performed on horses in Iran.

Experimental infection of embryonated eggs of chicken with Besnoitia caprae.

Knowledge on parasites of the genus Besnoitia, especially Besnoitia caprae, is sparse. Besnoitia caprae, an obligate intracellular protozoan parasite belonging to the phylum Apicomplexa, is the causative agent of caprine besnoitiosis. This experiment was conducted to determine the infectivity of the bradyzoites and the resultant histopathological lesions after inoculation of B. caprae bradyzoites in the embryonated egg. Eight groups, each having six embryonated eggs, were assigned in this experiment. Seven groups were inoculated with different doses of B. caprae bradyzoite inoculums (1x10(3), 1x10(4), 1x10(5), 1x10(6), 5x10(6), 1x10(7) and 2x10(7)) via the allantoic cavity route. The 8th group was considered as control. The embryos inoculated with higher doses showed mortality between 14 and 21 days of incubation (5-12 days post-infection). Those embryos that received lower doses were hatched on day 21 of incubation; however, they presented loss of feathers and paralysis and showed hyperemia in the skin of the feet regions. Histopathological sections of the skin revealed the presence of hemorrhages, hyperemia and inflammatory responses. Some of the chickens were euthanized after 50 days postinfection (DPI) and histopathological examination of their tissues revealed haemorrhages and coagulative necrosis with the presence of mononuclear cells infiltration in the liver and heart with interstitial pneumonia. It seems that the embryonated eggs could be a useful model to study the parasite's biology.