Normal anatomical and histochemical characteristics of the lacrimal glands in the American bison and cattle

Dorsal lacrimal glands, superior glands of the third eyelid and Harderian glands (deep gland of the third eyelid) from 19 bison and 18 cattle free of apparent ocular disease were examined to compare the normal anatomical properties of these glands. All glands were characterized and measured (length and width). The gross anatomy of the dorsal lacrimal glands was similar, with the exception of a bipartite gland in cattle. The bison's superior gland of the third eyelid and Harderian gland was longer as compared with cattle. A subset of the bison and cattle samples (five bison and five cattle) was sectioned for histological and histochemical analysis. The histology of the dorsal lacrimal and superior gland of the third eyelid revealed tubuloalveolar cells with basophilic vacuolated cytoplasm in bison and eosinophilic granular cytoplasm in cattle. The Harderian glands consisted of a tubuloalveolar anterior part combined with large lumens acini lined with cuboidal epithelium in the posterior part; the posterior part of the bison Harderian gland was more predominant than in cattle samples. Mucosubstance histochemistry revealed acidic and neutral glycoproteins with similar staining patterns in all glands of both species.     

Detection of lysozyme in llama, sheep, and cattle tears

OBJECTIVE: To determine whether the tears of llamas, sheep, and cattle contain lysozyme and compare lysozyme concentrations in tears among these species. ANIMALS: 40 llamas, 5 sheep, and 36 cattle. PROCEDURE: Electrophoresis, western blot immunoassay for lysozyme, a spectrophotometric assay to detect tear lysozyme by its ability to lyse a suspension of Micrococcus lysodeiticus, and a microtiter plate colorometric assay were performed. RESULTS: A 13.6-kd protein band was detected by use of electrophoresis and western blot immunoassay in llama and sheep tears but not cattle tears. Results of spectrophotometric assay suggested that llama and sheep tears had high concentrations of lysozyme, whereas cattle tears had low concentrations. Results of the microtiter plate colorometric assay suggested that llama tears had high concentrations of lysozyme, whereas concentrations in sheep and cattle tears were lower. CONCLUSIONS AND CLINICAL RELEVANCE: Lysozyme concentrations in tears may vary among species and this variability may contribute to differing susceptibilities to ocular diseases such as infectious keratoconjunctivitis.     

Evaluation of anti-Moraxella bovis pili immunoglobulin-A in tears following intranasal vaccination of cattle

Infectious bovine keratoconjunctivitis (IBK) is a highly contagious ocular disease of cattle caused by Moraxella bovis (Mb). Parenterally administered immunogens used to prevent the disease do not offer complete protection possibly because they stimulate a poor ocular mucosal secretory response, in which locally secreted immunoglobulin-A (sIgA) is one of the main components. The principal aim of this study was to evaluate by an indirect enzyme linked immunosorbent assay (ELISA), the local ocular mucosal sIgA response against Mb purified pili, produced after intranasal inoculation of experimental vaccines. Pili were adjuvanted by several different adjuvants (QuilA, Marcol Arlacel, Marcol Span, microencapsulated pili with PLGA polymers). Results were compared to sIgA response produced by adjuvant placebo inoculations and by IBK natural infection. Significantly higher anti-pili IgA response (p<0.05) was detected in calves vaccinated intranasally with pili QuilA and pili Marcol Span compared to control calves, although this specific immune response did not seem to be related to protection against Mb infection or typical IBK lesion development.     

Ultrastructural study of sheep lacrimal glands.

Sheep lacrimal glands are mixed glands, consisting of tubulo-acinar units succeeded by ducts of simple morphology. The secretory portions consist of three cell types: mucous, seromucous and serous, which may be intermingled in the same acinus or may form acini wholly made of only serous or mucous cells. Mucous cells show a rough endoplasmic reticulum that is reduced to a few cisternae located near the cell base and among the interstices of the secretory droplets. Mucous granules appear uniformly electron-lucent. Serous cells display a typical structure; serous granules can be uniformly electron-dense or composed of dense inclusions dispersed in an electron-lucent matrix. The seromucous granules have a bizonal substructure: a dense core is embedded in a highter matrix. Secretory acini are succeeded by intercalated ducts; the epithelium of these ducts gradually increases in height to form a kind of excretory duct, without the intervention of striated ducts.     

Normal ultrastructure and histochemical characteristics of canine lacrimal glands

Lacrimal glands of 12 dogs free of ocular disease were examined to determine the normal structure of these glands. The glands consisted of tubuloacinar cells that ultrastructurally and histochemically were of a single type of secretory cell in the tubules and possibly 3 types of secretory cells in the acini. The tubular epithelium contained homogenous electron-dense granules that stained as neutral glycoconjugates (periodic acid-Schiff positive and Alcian blue and high iron diamine negative). The predominant acinar cells contained granules of lesser electron density than those of the tubules, and stained as sialomucin (Alcian blue [pH 2.5] and periodic acid-Schiff-positive, and high iron diamine-negative). A second type of acinar cell was in peripheral lobules that ultrastructurally and histochemically appeared like lipid granules (positive with oil red O and osmium tetroxide). Ultrastructurally, a third type of acinar granule was finely granular, electron-lucent, and frequently coalesced. It was not readily apparent whether the latter was an artifact, a stage in the maturation of the sialomucin granules, or a third type of acinar granule. Individual acinar cells usually had a predominance of 1 granule type, but greater than 1 granule type could be found in some cells. The basal surfaces of the acinar, tubular, and ductal cells were incompletely ensheathed by myoepithelial cells. Plasma cells, lymphocytes, mast cells, endothelial cells, fat cells, and Schwann cells composed the cellular elements of the interstitium. Lymphocytes, mast cells, and nerve endings also were found in the parenchyma.     

Distribution of oxytetracycline into ocular tissues and tears of calves

A long-acting oxytetracycline formulation was administered (20 mg/kg of body weight) intramuscularly to calves, and the concentrations of the drug in serum, ocular tissues and tears were measured. The drug was distributed selectively to the epithelium of the conjunctiva and to the lacrimal gland ductules, and reached concentrations in each tissue that exceeded those in serum. The drug did not penetrate into the aqueous humour, and produced mean peak lacrimal fluid concentrations less than 1 microgram/ml after i.m. administration. When given subconjunctivally, however, concentrations greater than 2.0 micrograms/ml were observed in tears for 72 h. Severe local reactions occurred in all calves that were given the drug subconjunctivally.     

Multilocus sequence typing of Mycoplasma bovis reveals host-specific genotypes in cattle versus bison

Mycoplasma bovis is a primary agent of mastitis, pneumonia and arthritis in cattle and the bacterium most frequently isolated from the polymicrobial syndrome known as bovine respiratory disease complex. Recently, M. bovis has emerged as a significant health problem in bison, causing necrotic pharyngitis, pneumonia, dystocia and abortion. Whether isolates from cattle and bison comprise genetically distinct populations is unknown. This study describes the development of a highly discriminatory multilocus sequencing typing (MLST) method for M. bovis and its use to investigate the population structure of the bacterium. Genome sequences from six M. bovis isolates were used for selection of gene targets. Seven of 44 housekeeping genes initially evaluated were selected as targets on the basis of sequence variability and distribution within the genome. For each gene target sequence, four to seven alleles could be distinguished that collectively define 32 sequence types (STs) from a collection of 94 cattle isolates and 42 bison isolates. A phylogeny based on concatenated target gene sequences of each isolate revealed that bison isolates are genetically distinct from strains that infect cattle, suggesting recent disease outbreaks in bison may be due to the emergence of unique genetic variants. No correlation was found between ST and disease presentation or geographic origin. MLST data reported here were used to populate a newly created and publicly available, curated database to which researchers can contribute. The MLST scheme and database provide novel tools for exploring the population structure of M. bovis and tracking the evolution and spread of strains.     

A survey to detect Toxocara vitulorum and other gastrointestinal parasites in bison (Bison bison) herds from Manitoba and Saskatchewan

An egg count survey using environmental fecal samples obtained in spring or early summer was conducted to estimate the apparent prevalence of Toxocara vitulorum in unweaned bison calves and of other intestinal parasites in adult bison on 98 farms in Manitoba and Saskatchewan. Calf samples were pooled (maximum 5 samples per pool) by farm and positive pools were examined to determine individual T. vitulorum counts. Toxocara vitulorum eggs were found on 4 farms in Manitoba and none in Saskatchewan. Apparent herd-level prevalence estimates were 12% [95% confidence interval (95% CI): 3.4% to 28.2%] and 0% (95% CI: 0% to 5.7%) respectively. Samples from adult bison contained eggs/oocysts from trichostrongyle species, Eimeria sp., Monieza sp., Capillaria sp., Nematodirus sp. and Trichuris sp. in 100%, 95%, 72%, 13%, 13%, and 5% of herds, respectively. Strongyloides sp. were not found in any herd. Further studies are needed to assess parasite distribution patterns in bison and to evaluate the risk that T. vitulorum may pose to bison, cattle, and wildlife.     

夏季皖东牛与荷斯坦牛皮肤汗腺及其散热调节特征

为了探究夏季高温时皖东牛与荷斯坦牛皮肤组织形态和皮肤散热调节特征的差异,在30℃以上的夏日,选择皖东牛和荷斯坦牛母牛各4头,屠宰后采集颈部、肩部、背部、腹部、臀部和腿部6个部位的皮肤样本,通过HE染色切片分析皮肤的汗腺(SWG)与皮脂腺(SEG)的分布与形态,测定皮肤组织乙酰胆碱(ACh)浓度及乙酰胆碱转移酶(AChT)和酯酶(AChE)活力,采用RT-qPCR和Western blot检测皮肤组织乙酰胆碱M3受体(ACh M3R)、一氧化氮合酶(eNOs)及水通道蛋白5(AQP5)mRNA和蛋白的表达。结果:部位间比较,皖东牛与荷斯坦牛表皮厚度均为背部最厚,颈部最薄(P<0.05)。品种间比较,皖东牛平均皮肤厚度较高,平均表皮厚度较薄(P<0.05);汗腺密度与皮脂腺密度品种间无显著差异(P>0.05);皖东牛汗腺和皮脂腺的平均长度、直径、深度、体积均显著增加(P<0.05)。肩部、背部和臀部皮肤对比:夏季皖东牛和荷斯坦牛皮肤组织ACh浓度品种间无显著差异(P>0.05),部位间以臀部最高,肩部次之,背部最低(P<0.05)。ACh M3R和AQP...     

Evaluation of the presence of Leishmania spp. by real-time PCR in the lacrimal glands of dogs with leishmaniosis

Leishmania infantum infection is highly prevalent in endemic areas. Dogs with leishmaniosis may develop keratoconjunctivitis sicca (KCS). The goals of this study were (1) to quantify Leishmania amastigotes in the Meibomian glands (MG), main lacrimal gland (MLG) and nictitating membrane gland (NMG) from dogs with leishmaniosis; (2) to compare these results to immunohistochemistry (IHC), and (3) to explore the association between the Leishmania parasite load and the presence of ocular clinical signs. Twenty-five dogs diagnosed with leishmaniosis were included. MG, MLG and NMG from both eyes were collected. Histopathology, IHC and real-time PCR were performed. All specimens yielded positive real-time PCR results. For all three glands, samples from dogs with ocular clinical signs had mean ΔCt (cycle threshold) values significantly lower (higher parasite loads) than those from dogs without signs. Cut-off values of ΔCt<0, ΔCt<4 and ΔCt<4.9 for MG, MLG and NMG, resulted in a likelihood ratio of positives of 5.9, 6.38 and 6.38, respectively. Samples with ΔCt values below the reported cut-off were significantly more likely to display clinical signs related to KCS than those with results above the cut-off, for all three glands. Similarly, ΔCt values below the cut-off were significantly associated with positive IHC. In this study real-time PCR has been standardised for use in MG, MLG and NMG. A cut-off value established for each of these tissues may aid the clinician in the discrimination between ocular signs related to Leishmania from those associated with other causes of KCS.     

Distribution of cholinergic and adrenergic nerve fibers in the lacrimal glands of dogs

The distribution of postganglionic autonomic nerve fibers in the lacrimal gland and gland of the third eyelid of dogs was studied by use of histochemical techniques. Adrenergic nerve distribution was identified by use of the sucrose-potassium phosphate-glyoxylic acid technique. A loose network of adrenergic nerves was found throughout the interstitium around acini and blood vessels and in vessel walls. Acetylcholinesterase staining was used to identify cholinergic nerve fibers. A cholinergic distribution pattern was found, although the cholinergic innervation appeared more dense than the adrenergic. In the gland of the third eyelid, mucus-secreting lobules and lipid-secreting lobules appeared to be equally innervated by parasympathetic fibers. These lobules could not be differentiated when the sucrose-potassium phosphate-glyoxylic acid technique was used. The techniques used in this study could not demonstrate whether direct contact was made by either cholinergic or adrenergic nerve fibers with secretory or myoepithelial cells. The presence of both nerve fiber types around acini suggests an interrelationship between the sympathetic and parasympathetic nervous system in lacrimal gland secretion in dogs.     

Immobilization of cattle and bison with a combination of xylazine, zolazepam-tiletamine and ketamine

The aim of this study was to find a safe and reliable alternative to Immobilon for the immobilization of (feral) cattle. A combination of xylazine, zolazepam-tiletamine and ketamine was tested in Limousin cattle, Scottish Highland cattle, and American bison. Bodyweight, induction time, arterial O2 saturation and the total downtime were measured. Arterial blood was taken for pH and blood gas analysis. The animals were then injected with atipamezole and the recovery time was recorded. A combination of 500 mg zolazepam, 500 mg tiletamine, 500 mg xylazine, and 1000 mg (10 ml) ketamine, administered in a dosage of 1 ml per 100-150 kg bodyweight (depending on the species), proved to be most reliable and effective. The combination resulted in a fast immobilization. In all animals slight respiratory depression was seen, which indicates that oxygen suppletion may be needed for long-lasting immobilization. After reversal of the xylazine component, almost all animals recovered within 4 minutes. No long term adverse effects were reported by the owners.     

Lysozyme concentrations in the tears of cattle, goats, and sheep

Tear samples were collected from 1 eye of each of 40 cows, 27 sheep, 5 goats, and 5 human beings. Additionally, 10 bovine tear samples were pooled and concentrated. Spectrophotometric assays, using Micrococcus lysodeikticus, were performed on each sample to detect lysozyme activity expressed in hen egg lysozyme (HEL) equivalents. Lysozyme activity was not detected in tears of cows, but 158.8 +/- 159.3 mg of HEL/ml was detected in tears of sheep, 220.7 +/- 37.5 mg of HEL/ml in tears of goats, and 216.3 +/- 86.2 mg of HEL/ml in tears of human beings. In pooled bovine tear samples, lysozyme activity was not detected on plate assay and lysozyme protein was not detected on polyacrylamide gel electrophoresis, column chromatography, or immunoelectrophoresis with rabbit anti-bovine tear antibodies. On the basis of these observations, we concluded that the basic ocular protective mechanism in bovine tears is not lysozyme. Other anti-bacterial proteins such as lactoferrin, transferrin, complement, or beta-lysin may, therefore, be of primary importance in protecting the bovine eye.     

The inheritance of ocular colobomata in Charolais cattle

The examination of approximately 800 pure bred and cross bred Charolais cattle indicated that ocular colobomata is inherited via an autosomal dominant gene in this breed. Penetrance was found to be complete in the male and partial (52 per cent) in the female. Inheritance in the crossbred animals was different to that in the pure bred.     

Development and stability of rinderpest virus antigens in cattle tears and lymph nodes

Summary Diffusible rinderpest virus antigens were demonstrated in increasing quantities in ocular and lymph node biopsies from rinderpest-infected cattle using agar gel immunodiffusion (AGID) and counterimmunoelectrophoresis (CIEP) tests. Positive samples were detected from the second day of pyrexia to two days after death. The antigens in ocular secretions and lymph nodes were thermolabile being destroyed within five minutes at 56°C and within two weeks at 4°C.

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Resumen Se demostró un incremento de antígenos difusibles del virus de rinderpest, en biopsias oculares y linfáticas tomadas de ganado inoculado experimentalmente. Las pruebas utilizadas fueron la inmunodifusión en agar gelatina y la contrainmunoelectroforesis. Se detectaron muestras positivas desde el segundo día de pirexia, hasta dos días antes de la muerte. Los antígenos en las secreciones oculares y nódulos linfáticos fueron termolábiles, siendo destruidos a los cinco minutos a 56°C y en dos semanas a 4°C.

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Résumé Les tests d’immuno-diffusion en gélose (IDG) et de contre-immunoélectrophorèse (CIE) ont permis de mettre en évidence des quantités croissantes d’antigènes diffusibles du virus de la peste bovine dans les prélèvements de larmes et de ganglions lymphatiques. Des résultats positifs ont été enregistrés à partir du deuxième jour de la fièvre jusqu’au 2e jour après la mort. Les antigènes des secrétions oculaires et des ganglions lymphatiques sont thermolabiles: destruction en cinq minutes à 56°C et en deux semaines à 4°C.

     

Malignant catarrhal fever: polymerase chain reaction survey for ovine herpesvirus 2 and other persistent herpesvirus and retrovirus infections of dairy cattle and bison.

Using a polymerase chain reaction (PCR) test for sequences of ovine herpesvirus 2 (OHV2), this virus was shown to be significantly associated with sheep-associated malignant catarrhal fever (SA-MCF) in terminal cases of disease in 34 cattle and 53 bison. Ovine herpesvirus 2 was not detected in cattle (38) and bison (10) that succumbed to other diseases. Other persistent herpesviruses, retroviruses, and pestivirus, some of which have been previously isolated from cases of SA-MCF, were not associated with the disease. These included bovine herpesvirus 4 (BHV4), bovine lymphotrophic herpesvirus (BLHV), bovine syncytial virus (BSV, also known as bovine spumavirus), bovine immunodeficiency virus (BIV), and bovine viral diarrhea virus (BVDV). A PCR survey for OHV2 in DNA from individual cow's peripheral blood lymphocytes in 4 dairies showed that the 1 dairy that was in close contact to sheep had a prevalence of OHV2 of 21.3%, whereas the 3 other dairies had no OHV2. Prevalence of the other herpesviruses and retroviruses in the dairy cows was variable, ranging from 2% to 51% for BHV4, 52% to 78.7% for BLHV, and 10% to 34% for BSV. Bovine lymphotrophic herpesvirus and BSV were also found in a few (1-4 of 21 tested) cases of terminal SA-MCF, but BIV and BVDV were not found in either the dairy cows sampled, or in the cases of SA-MCE No significant correlation was found between the presence of any 2 viruses (OHV2, BHV4, BLHV, BSV) in the dairy cows or terminal cases of SA-MCE     

Evidence for papillomaviruses in ocular lesions in cattle

Negatively stained preparations of various ocular lesions, generally considered to be precursors to bovine ocular squamous cell carcinoma, were subjected to electron microscopic examination for viruses. Lesions examined included five conjunctival plaques, an acanthotic lesion from an eyelid, 15 conjunctival papillomas, two papillomas of the eyelid and two keratinised elongated proliferative lesions of the eyelid (cutaneous horns). Eight lesions contained particles that resembled papillomaviruses. These consisted of a conjunctival plaque, five conjunctival papillomas, a papilloma of the eyelid and one of the samples of cutaneous horn. The particles were present in small numbers and were found only after prolonged search. The finding of these particles suggests that papillomavirus may be involved in the aetiology of bovine ocular squamous cell carcinoma.     

静止期与活动期黄牛乳腺肥大细胞的分布规律

乳腺炎是奶牛最重要的感染性疾病之一 ,目前 ,对于本病的防治 ,国内外学者采用了多种措施 ,但都未能取得满意的疗效。 Sordilo等曾用白介素 - 2 (IL-2 )对奶牛乳腺炎进行治疗研究 ,但由于迄今为止尚不清楚 IL- 2与乳腺炎发病机制的关系 ,其疗效也不理想[1] 。人和鼠类肥大细胞在免疫复合物 (immunecomplexes)、寄生虫和细菌等诱发的炎症反应中发挥重要作用[2 ] 。但对牛肥大细胞在乳腺炎的免疫学作用尚不清楚。Nielsen发现泌乳后期牛乳腺小叶间肥大细胞内有 4种不同形态的膜包颗粒 ,与人和某些实验动物的肥大细胞十分相似[3 ] 。但肥大细…     

Heavy sarcocystosis in the ocular musculature of cattle and buffaloes

A high prevalence of 71.5 per cent and 69.7 per cent of sarcocystosis was observed in the ocular musculature of cattle and buffaloes respectively, in Bihar, India. The concentration of cysts in the eye muscle was also usually heavy. Ocular musculature appears to be a preferred site for the development ofSarcocystis in these intermediate hosts, second only to the heart muscle. The species ofSarcocystis involved in the present study were morphologically indistinguishable fromS. cruzi in cattle andS. levinei in buffaloes. This appears to be the first report on the occurrence ofS. cruzi andS. levinei in ocular musculature.