Immunopathogenesis of keratoconjunctivitis sicca in the dog.

Keratoconjunctivitis sicca (KCS), more commonly known as dry eye, is an inflammatory condition of the ocular surface caused by a pathologic reduction in the aqueous component of the tear film. It is seen commonly in the dog and defined as a Schirmer tear test with a reading of less than 10 mm in one minute. While KCS may be caused by neurological disease or drug toxicity, most cases are immune-mediated. Whereas the immunological basis of autoimmune KCS has been extensively investigated in humans and experimental rodent models, little research has been undertaken in the dog. It is hoped that this review spurs further research into the etiopathogenic factors in canine KCS.     

Tear-deficient and evaporative dry eye syndromes of the horse

Tear-deficient dry eye (keratoconjunctivitis sicca), and evaporative dry eye, with some overlap between these two categories, are two major categories of dry eye recognized in the horse. Careful examination of the eyelid margins, the inner and outer surfaces of the upper and lower eyelids, assessment of blink and third eyelid movement, specific testing of tear production, assessment of corneal sensitivity, and slit-lamp examination of the ocular surface before and after fluorescein dye application is recommended to detect dry eye problems. Rose Bengal dye application is also recommended in many cases. Facial nerve paresis is a possible complicating factor in some instances. Evaporative dry eye deserves closer scrutiny for it may be a contributory factor when ocular surface disease such as keratomycosis is present. Factors which influence ocular surface health need more close examination in horses.     

Qualitative tear film disease

Abnormalities of the lipid and mucin components of the preocular tear film may result from diseases of the eyelid margins and conjunctiva. Chronic keratoconjunctivitis with epithelial edema and superficial corneal neovascularization, with or without ulceration, characterizes qualitative tear diseases. Tear components other than lipid and mucin that carry probable clinical significance include tear proteins, all-trans retinal, cholesterol, glucose, and electrolytes. Although less common than quantitative or aqueous deficiencies, qualitative abnormalities are recognized as primary or secondary causes of ocular surface disease in companion animals.     

Acidification of the colonic mucins following polyvalent colonization of the germ-free rat.

Colonic mucins of germ-free (GF) and conventional rats (CV) were compared. After isolation by gel filtration on Sepharose CL-4B and purification by density gradient centrifugation, the content of isolated colonic mucins was estimated by determination of PAS positive carbohydrates. Purified mucins were subjected to carbohydrate and amino acid analysis and separated into mucin subclasses by ion exchange chromatography. While the total amount of colonic mucins was not statistically different in GF and CV animals, analysis of carbohydrate composition demonstrated an increased amount of sialic acid in CV rat mucin. This was in accordance with results of ion exchange chromatography, revealing a significant higher amount of negative charged mucin subclasses in CV mucin, compared to the germ-free counterpart. The results of amino acid analysis were similar in both groups. The compositional differences in carbohydrate moieties are attributed to modulations by the intestinal flora. A selective bacterial degradation of the neutral mucin subclasses and modifications in the mucin composition due to a stimulated synthesis are discussed.     

Ocular parameters related to drug delivery in the canine and equine eye: aqueous and vitreous humor volume and scleral surface area and thickness

OBJECTIVE: To measure the ocular surface area, scleral thickness, and the aqueous and vitreous humor volumes in the canine and equine eye. Animals studied Fourteen canine and 16 equine cadaver eyes. PROCEDURE: Eyes were measured either fresh or following fixation in formalin. Ocular surface area was calculated using the fluid displacement method. Globes were hemisected and aqueous and vitreous humor were collected and quantitated. Scleral thickness was measured by digital caliper, by image projection, and by ultrasonic biomicroscopy (UBM). RESULTS: Mean +/- standard deviation (SD) scleral surface areas in canine and equine eyes were 12.87 +/- 2.24 and 57.23 +/- 5.63 cm2, respectively. Mean +/- SD aqueous humor volume was 0.77 +/- 0.24 in the dog and 3.04 +/- 1.27 mL in the horse. Mean vitreous humor volume was 1.7 +/- 0.86 mL for the canine eye and 26.15 +/- 4.87 mL for the equine eye. In canine and equine eyes, the sclera was thinnest at the ocular equator: 0.34 +/- 0.13 mm (canine) and 0.53 +/- 0.1 (equine). There were no significant differences between the direct caliper and projected thickness measurements or differences between measurements in the superior, inferior, nasal, and temporal quadrants of the eye. Scleral thickness in fresh tissue was greater than in fixed tissue at most sites. CONCLUSIONS: The UBM measurement method appeared to be most consistent and reproducible when compared to direct measurement techniques. Formalin fixation of the eyes was associated with significantly thinner scleral tissue than with fresh ocular tissue.     

Expression of secreted mucins (MUC2, MUC5AC, MUC5B, and MUC6) and membrane-bound mucin (MUC4) in the lungs of pigs experimentally infected with Actinobacillus pleuropneumoniae

The expression patterns of different secreted (MUC2, MUC5AC, MUC5B, and MUC6) and membrane-bound (MUC4) mucins were determined immunohistochemically in the lungs of pigs experimentally infected with Actinobacillus pleuropneumoniae. Forty-seven-week-old colostrum-deprived pigs were randomly allocated to infected (n=20) or control groups (n=20). Five infected and uninfected pigs were euthanized at 0, 6, 12, and 48 h post-inoculation (hpi). In the infected pigs, the expression of both types of mucins, which were invariably observed, was associated with bronchiolar and respiratory bronchiolar lesions. Strong positive mucin signals were seen on the surface of bronchiolar and respiratory bronchiolar epithelium with neutrophil infiltration. The mean mucin-positive area peaked at 6 hpi and decreased significantly to control levels by 48 hpi on the surface of the bronchiolar and respiratory bronchiolar epithelium. Further studies are needed to establish the functional relationship between mucin expression and the host defense mechanism against A. pleuropneumoniae in the lungs of infected pigs.     

Efficacy of topically applied liposome-bound tetracycline in the treatment of dry eye model

Objective To evaluate the effects of liposome‐bound tetracycline eye drops in a rabbit dry eye model evaluating their advantage of being less allergic, preservative free and prolonged action compared with other tear substitutes. Procedures New Zealand albino rabbits were equally divided into control group and dry eye induced groups. Dryness was induced in 24 eyes of 12 healthy adult male albino rabbits by instilling atropine sulfate eye drops 1% three times daily for 1 week, then animals were subdivided into four groups; group 1 (rabbits with dry eye model), groups 2, 3, and 4: rabbits with dry eye model treated for 7 days starting on 7th day of dryness induction with either tetracycline, empty liposome, or combined tetracycline with liposome as topical eye drops respectively. Schirmer (STT) test and tear break up time (TBUT) were assessed on days 0, 2, 4, 7, 9, 11, and 14. Animals were sacrificed on day 14 and histopathological examination of the cornea and conjunctiva was performed. Results Tear break up time and STT test values were significantly improved in groups 2, 3, 4 as compared with group 1. The histopathological examination showed normal cytoarchitecture of corneas and conjunctivae in groups 2, 3, 4 against the dryness effect that continued to affect the cornea and conjunctival epithelium in group 1. There was a significant improvement in the group treated with liposome‐bound tetracycline eye drops (group 4) as compared with tetracycline alone (group 2) and empty liposome (group 3). Conclusion The use of liposome encapsulated tetracycline significantly improved STT and TBUT values as well as reverse surface ocular pathology.     

Expression of secreted and membrane-bound mucins in the airways of piglets experimentally infected with Mycoplasma hyopneumoniae

Immunohistochemistry was used to demonstrate secreted mucins MUC2, MUC5AC and MUC5B and membrane-bound mucin MUC4 in the pulmonary bronchioles of piglets experimentally infected with Mycoplasma hyopneumoniae. Conventional status, Landrace-Duroc cross-bred piglets, 13 days of age, were randomised to two groups. One group (n=20) was infected by the intra-tracheal route with the SNU98703 strain of M. hyopneumoniae, and a group of 12 animals acted as uninfected controls. Five infected and three uninfected piglets were euthanased on the day of infection and at 7, 21, and 35 days post-inoculation (PI). Membrane-bound MUC4 and secreted MUC5AC were the predominant mucins produced in the bronchioles of the piglets in response to M. hyopneumoniae infection, but by day 35 PI, all labelled mucins had returned to pre-infection levels, contemporaneous with reduced pulmonary lesion scores. The increased mucin production may result from direct stimulation of the epithelium by mycoplasmal infection, or may arise indirectly following M. hyopneumoniae-induced ciliostasis.     

Morphological,morphometric and histochemical characterization of the gastric mucosa of the camel (Camelus dromedarius)

Mucosal morphology, morphometry and mucin histochemistry of the stomach were studied in the one-humped camel. The lining of the stomach was divided into eight grossly identifiable regions. The first region was non-glandular, occupied the body of the first compartment of the stomach and constituted 53.2% of the gastric mucosa. The other seven regions were lined by a glandular mucosa. Histological, histochemical and morphometric investigations have shown that glandular mucosa comprises pseudo-cardiac, cardiac, fundic and pyloric regions. The pseudo-cardiac region was characterized by widely separated short tubular serous glands. It constituted 36.2% of the gastric mucosa; it extended over the entire lining of the second compartment and parts of the first and third compartments. The cardiac region was confined to the initial zone of the third compartment amidst the psuedo-cardiac region but contiguous with the distal end of the gastric groove. It constituted 3.4% of the gastric mucosa and was characterized by neutral and acid mucin positive glands. The fundic and pyloric regions occupied the distal distended part of the third compartment. The fundic region constituted 4.3% of the gastric mucosa. It was packed with typical fundic glands characterized by chief cells, parietal cells and acid mucin positive neck cells. The pyloric region constituted 2.9% of the gastric mucosa. Its glands were positive to acid mucins except for their bases that were positive to neutral mucins. Differences in volume densities of the mucosal components and reactivity of the surface epithelium and gastric pits to mucin stains were noted in the different regions of the stomach.     

Mucin biosynthesis in the bovine goblet cell induced by Cooperia oncophora infection

Mucin hypersecretion is considered to be one of the most common components of the immune response to gastrointestinal nematode infection. However, investigations have not been conducted in the Cattle–Cooperia oncophora system to verify the findings largely derived from murine models. In this study, we examined the expression of seven mucins and seven enzymes in the mucin biosynthesis pathway involved in O-linked glycosylation in the bovine small intestine including goblet cells enriched using laser capture microdissection during a primary C. oncophora infection. At the mRNA level, MUC2 expression was significantly higher in both lamina propria and goblet cells at 28 days post-infection compared to the naïve control. MUC5B expression at the mRNA level was also higher in lamina propria at 28 dpi. Expression of MUC1, MUC4, MUC5AC, and MUC6 was extremely low or not detectable in goblet cells, columnar epithelial cells, and lamina propria from both naïve control and infected animals. Among the seven enzymes involved in post-translational O-linked glycosylation of mucins, GCNT3, which may represent one of the key rate-limiting steps in mucin biosynthesis, was up-regulated in goblet cells, columnar epithelial cells, lamina propria, and gross small intestine tissue during the course of infection. Western blot analysis revealed that MUC2 glycoprotein was strongly induced by infection in both gross small intestine tissue and its mucosal layer. In contrast, the higher MUC5B protein expression was observed only in the mucosal layer. Immunohistochemistry provided further evidence of the mucin glycoprotein production and localization. Our results provided insight into regulation of mucin biosynthesis in various cell types in the bovine small intestine during gastrointestinal nematode infection and will facilitate our understanding of mucins and their role in immune response against parasitic nematodes.     

Refined methodology to purify mucins from pig colonic mucosa

Mucin, the main macromolecular component of mucus, contains a peptide backbone rich in proline, threonine and serine (PTS) and oligosaccharide side chains. Glycosylation increases the molecular size of mucins and restricts access of proteases to mucin. Based on these characteristics, Faure et al. [Faure, M., Moënnoz, D., Montigon, F., Fay, L.B., Breuille, D., Finot, P.A., Ballèvre, O. and J. Boza. 2002. Development of a rapid and convenient method to purify mucins and determine their in vivo synthesis rate in rats. Anal. Biochem 307: 244–251.] developed a method to purify mucins from intestinal mucosa. In this method, mucins are reduced (RD) and digested with protease (dig) prior to isolation using size-exclusion chromatography. Our objective was to refine this method to purify mucins from pig colonic mucosa. Mucosal scrapings were homogenized in 5 mM EDTA and were either treated with 7 μL of protease inhibitor cocktail (PIC; UndigUnRD) or incubated with 100 μL of Protease at 37 °C for 80 min and then subsequently treated with PIC (digUnRD). Homogenates were fractionated based on size using a Sepharose CL-4B column. Mucin-containing fractions were identified by a combination of SDS-PAGE and staining techniques and were pooled. Mucin purified using digUnRD method had higher proportions of PTS compared to crude mucin purified using UndigUnRD method (31.9 vs. 23.5 mol/100 mol AA; P < 0.005). The proportion of PTS in mucin purified using digUnRD was also higher than mucin purified using digRD (31.9 vs. 25.7 mol/100 mol AA; P < 0.05). The refined method employing protease digestion and no reduction can be successfully applied to purify mucins from pig colonic mucosa.     

黏蛋白侧链岩藻糖在肠道细菌与宿主互作过程中的作用

消化道表面黏液层中含有大量黏蛋白,这些黏蛋白分子侧链往往被糖基化修饰,这些糖基在肠道菌的黏附、定植及免疫调节过程中发挥重要作用。岩藻糖是肠道黏蛋白分子侧链中的一种重要糖基,本文总结了岩藻糖作为肠道菌黏附靶位点和信号调节分子,以及调节肠道功能等方面的作用。     

Sheep intelectin-2 co-purifies with the mucin Muc5ac from gastric mucus

Secretion of gastric mucins plays an essential role in host protection, and modifications in mucus properties are characteristic of the protective immune responses to pathogens. This study describes the purification and characterisation of sheep gastric mucins, and identification of those proteins that co-purify with mucins, with the potential to modify mucus properties. Gastric mucus was collected and pooled from four abattoir sheep and separated by CsCl density gradient centrifugation. Proteomic analysis of the mucin-containing fraction indicated the presence of gastric mucin (Muc5ac) and several co-purifying proteins, including intelectin-2 (Itln2). Further experimentation indicated that a combination of denaturation and reduction was required to fully release Itln2 from gastric mucin. A putative correlation was found between mucin-bound intelectin concentration and rheological properties in further sheep gastric mucus samples. In conclusion, this study provides the first characterisation of sheep gastric mucins and their purification partners, revealing potentially important mucin-intelectin interactions.     

Sheep intelectin-2 co-purifies with the mucin Muc5ac from gastric mucus

Secretion of gastric mucins plays an essential role in host protection, and modifications in mucus properties are characteristic of the protective immune responses to pathogens. This study describes the purification and characterisation of sheep gastric mucins, and identification of those proteins that co-purify with mucins, with the potential to modify mucus properties. Gastric mucus was collected and pooled from four abattoir sheep and separated by CsCl density gradient centrifugation. Proteomic analysis of the mucin-containing fraction indicated the presence of gastric mucin (Muc5ac) and several co-purifying proteins, including intelectin-2 (Itln2). Further experimentation indicated that a combination of denaturation and reduction was required to fully release Itln2 from gastric mucin. A putative correlation was found between mucin-bound intelectin concentration and rheological properties in further sheep gastric mucus samples. In conclusion, this study provides the first characterisation of sheep gastric mucins and their purification partners, revealing potentially important mucin-intelectin interactions.     

Ocular infection of donkeys (Equus asinus) with <Emphasis Type="Italic">Setaria equina</Emphasis>

Seven donkeys raised in different locations in Egypt were found to have a unilateral eye showing motile white worms in the aqueous humor. The parasites were surgically removed from the anterior chamber of the eye in five out of the seven donkeys, and were cleared, mounted and identified as Setaria equina based on light microscopic features. The ocular infection with S. equina reported herein may be the first reported aberrant cases in the Egyptian animals.     

Cosmetic globe surgery in the horse.

Effects of traumatic injury or inflammation on the equine eye can be catastrophic. These ocular conditions can frequently result in blindness or chronic pain. In addition to blindness and pain, permanent unsightly cosmetic defects can occur. This article addresses options available for improved cosmetic outcome in horses with ocular scars or requiring enucleation. Many of these options have been described in detail previously. New information has been added to the discussion of each option where pertinent.     

Ocular infection of cattle with Setaria digitata

One 5-month-old female native Korean calf and a 2-year-old female Holstein cow raised in two farms about 4 km apart from each other in Korea, were found to have the left eye opaque, which included motile white worms in the aqueous humor. The parasite removed from the left eye of the calf was identified as Setaria digitata based on both light and electron microscopic features. The ocular infection with S. digitata reported herein may document the first aberrant case in Korean cattle.     

Corneal concentrations and preliminary toxicological evaluation of an itraconazole/dimethyl sulphoxide ophthalmic ointment

The objectives of this study were to determine the concentration of itraconazole achieved in corneal tissue and aqueous humour after topical application of a 1% itraconazole ointment; to determine the effect of including dimethyl sulphoxide (DMSO) in the ointment on achievable ocular tissue itraconazole concentrations; and to assess if any gross or histopathologic ocular toxicity results from the topical application of 1% itraconazole with or without the addition of DMSO.
The experimental trial consisted of 6 horses considered to have normal eyes. Each horse had one eye treated with 0.3 mL of 1% ultra-micronized itraconazole ointment and the fellow eye with 0.3 mL of 1% itraconazole/30% DMSO ointment. The ointment was applied every 6 h for a total of 28 treatments. Both ointments were well tolerated and no gross or histopathologic abnormalities developed during the trial.
Corneal tissue and aqueous humour concentrations of itraconazole were determined using high performance liquid chromatography. Corneal tissue concentration averaged 1.1 (± 0.4) μg/g in horses treated with the 1% ultra-micronized itraconazole ointment and 7.9 (± 3.3) μg/g for those treated with the 1% itraconazole/30% DMSO ointment; there was a statistically significant difference between ointments ( P = 0.005) No itraconazole could be detected in the aqueous humour in either treatment group.     

The Prenatal Development and Histochemistry of the Ileal Mucins in the Bovine Fetuses

Few studies exist regarding the distribution of intestinal mucins in fetuses of mammalians such as cattle and sheep. In this study, we aimed to describe the changes in the mucin production by ileal epithelium of bovine fetuses during their prenatal development. The goblet cells showed heterogeneity in mucins and the apical cytoplasm of the enterocytes demonstrated Periodic acid Schiff‐positive reaction which declined gradually towards the birth. Moreover, the number of the goblet cells containing acidic and mixed mucins augmented, whereas those containing neutral mucins decreased with advancing gestational age. After sixth month of gestation, with the initiation of the ileal Peyer patches and follicle‐associated epithelium development, a gradual increase in the number of goblet cells containing sulfomucins was also noticed towards the birth. The presence of different mucins in the ileum of bovine fetuses throughout prenatal development might play a role in the protection of the intestinal mucosa against urinary waste products in swallowed amniotic fluid and bile. Furthermore, mucins can also contribute for the formation of meconium in intra‐uterine life and building of strong intestinal barrier with predominating sulfomucins, protecting the intestine against potential pathogens and digestive enzymes after birth.