In vivo confocal microscopy of equine fungal keratitis

Objective To describe in vivo corneal confocal microscopy of horses with fungal keratitis and correlate findings with clinical, histopathological, and microbiological evaluations of clinical cases and an ex vivo experimental equine fungal keratitis model. Animals studied A total of 12 horses with naturally‐acquired fungal keratitis and ex vivo equine corneas experimentally infected with clinical fungal isolates. Procedures Horses with naturally‐acquired fungal keratitis were examined with a modified Heidelberg Retina Tomograph II and Rostock Cornea Module. Confocal microscopy images of clinical isolates of Aspergillus fumigatus, Fusarium solani, and Candida albicans were obtained by examination of in vitro cultures and experimentally infected ex vivo equine corneas. Results Non‐specific in vivo corneal confocal microscopic findings in horses with fungal keratitis included leukocyte infiltrates, activated keratocytes, anterior stromal dendritic cell infiltrates, and vascularization. Linear, branching, hyper‐reflective structures that were 2–6 μm in width and 200 to >400 μm in length were detected in all horses with filamentous fungal keratitis. Round to oval hyper‐reflective structures that were 2–8 μm in diameter were detected in a horse with yeast fungal keratitis. The in vivo confocal microscopic appearance of the organisms was consistent with fungal morphologies observed during examination of in vitro cultures and infected ex vivo equine corneas. Conclusions In vivo corneal confocal microscopy is a rapid and non‐invasive method of diagnosing fungal keratitis in the horse. This imaging technique is useful for both ulcerative and non‐ulcerative fungal keratitis, and is particularly advantageous for confirming the presence of fungi in deep corneal stromal lesions.     

AM真菌对紫花苜蓿应答蚜虫胁迫的影响

紫花苜蓿是优质的多年生豆科牧草,广泛种植于世界各地。豌豆蚜是紫花苜蓿的主要害虫之一,其不仅可危害植物生长,还会传播多种植物病害,从而降低牧草产量和品质,造成巨大的经济损失。丛枝菌根(AM)真菌可与80%的陆生植物根系共生形成互惠共生的菌根结构,其可促进植物对矿物营养的吸收,增强植株抗逆性。以紫花苜蓿为材料,探究AM真菌对植物应答豌豆蚜胁迫的生化机理。结果表明,AM真菌可以通过促进植物生长和养分吸收,改变植株防御性酶活性以及植物激素信号物质含量来调控植物自身对蚜虫的响应。2种AM真菌根内球囊霉和幼套球囊霉均显著提高了紫花苜蓿生物量、分枝数和植株N、P含量(P<0.05)。此外,根内球囊霉显著提高了植物过氧化物酶(POD)活性,且蚜虫处理下,较不接菌处理,根内球囊霉植株的POD、过氧化氢酶(CAT)活性以及激素信号物质水杨酸(SA)含量均显著增加(P<0.05)。在蚜虫处理下,相比不接菌处理,幼套球囊霉显著增强了植株POD活性(P<0.05)。蚜虫取食显著提高了植株SA含量,降低了超氧化物歧化酶(SOD)活性(P<0.05);且接种AM真菌后,蚜虫处理下的根系POD活性和SA含量提升程度更大,表明AM真菌可抑制蚜虫对植株造成的负效应。     

AM真菌提高植物抗逆性的机制

丛枝菌根(arbuscular mycorrhizae,AM)真菌是自然界中最广泛的植物共生真菌,可以提高宿主对生物和非生物逆境的抗性。AM真菌能通过改善养分吸收、增加渗透调节物质的积累,提高抗氧化酶类活性、强化渗透调节作用并维持植物内源激素平衡,增加生长素的合成,调节碳氮代谢,刺激胁迫诱导基因表达,增强植物根系及菌丝本身对重金属元素的固持作用等提高植物对非生物逆境(如干旱、高低温、重金属、盐碱)的抗性;并能通过构建菌丝网络,形成对根部入侵病原真菌的机械屏障,增强抗病性相关酶活性,合成与抗病性相关的次生代谢产物,增强抗病、虫相关基因的表达,菌丝传递抗虫防御信号,能提高邻近植株的抗虫性,增强植物的抗病虫能力。本文归纳了近几年来国内外有关AM真菌影响植物抗逆性及其作用机理的研究进展,并对AM真菌在促进植物抗逆性方面的研究作了展望。     

Immunology of fungal infections in animals

The nature of immunity to fungal infection is discussed predominantly for mammals and birds. T-cell-mediated immunity seems essential for recovery both from cutaneous and mucosal infections (Candida, Malassezia and dermatophytes) and from infections of systemic fungal pathogens (Cryptococcus, Blastomyces, Histoplasma, and Coccidioides). Often chronic progressive disease caused by these fungi is associated with a depression or absence of T-cell-mediated immunity to antigens of the infecting fungus. In contrast recovery from disease, or absence of clinical disease after exposure to these fungi, is associated with the presence of strong T-cell-mediated immune responses to the fungus. The activation of macrophages and the stimulation of epidermal growth and keratinization are the processes induced by T-cell-mediated immunity which result in the resolution of systemic or cutaneous and mucosal disease. Other cell types, for example NK cells and PMNs (polymorphonuclear leucocytes), may be important in these diseases in reducing the effective amount of inoculum to which an animal is exposed and thereby reducing the likelihood of disseminated disease. Invasive opportunistic fungi (Candida, Aspergillus, Mucorales) are resisted by PMNs which attach to the hyphae or pseudohyphae and damage them via an extracellular mechanism. Other host cell types may be important in natural resistance, fungal spores being handled by the macrophages which, under conditions when animals are not immunosuppressed, are likely to be an effective first line of defense. Subcutaneous pathogens and miscellaneous other fungal diseases are discussed from a point of view of host immunity and immunodiagnosis. Vaccine development for ringworm and for other mycoses is discussed.     

P‐77 Skin fungal flora in horses: is Malassezia an important component

The aim of this study was to evaluate the fungi present on the skin surface in healthy horses with a particular interest in Malassezia on predisposed skin areas. Twenty‐five mares were included, each sampled on nine skin areas including those most likely contaminated by Malassezia: nostrils, lips, ear pinnae, base of the mane, pasterns, pectoral areas, lateral thorax, udder and perianal areas. Two sterile carpets were applied to the skin, and then used for inoculation of agar (Sabouraud's plus cycloheximide and chloramphenicol), both with and without lipids for isolation of different species of Malassezia. This method is currently successful in isolating Malassezia in dogs. Cultures were incubated at 32°C, observed on day 5 for Malassezia, and then maintained at 27°C for complementary morphological identifications. From 223 cultures on standard medium, only seven remained sterile. The most common fungi were filamentous (number of positive samples): Scopulariopsis (156), Penicillium (79), Acremonium (67), Aspergillus (53), Paecilomyces spp. (38), and Stachybotrys (33). Interestingly, Microsporum gypseum was also isolated (14 samples from seven areas) from 10 horses. Yeast or yeast‐like fungi were Geotrichum (52), Trichosporon (21), and an additional 33 samples being positive for yeast other than Malassezia. Results from the other 223 cultures on lipid‐rich agar were similar. Most of the fungi were isolated from the nine areas selected, but Malassezia was never isolated from 446 cultures. Funding: Self‐funded.     

Immune synergistic oligodeoxynucleotide from Lactobacillus rhamnosus GG enhances the immune response upon co‐stimulation by bacterial and fungal cell wall components

Bacterial genomic DNA has recently been shown to elicit a highly evolved immune defense. This response can be selectively triggered for a wide range of therapeutic applications, including use as a vaccine adjuvant to immunotherapies for allergy, cancer, and infectious diseases. Previously, we identified a low‐concentration immune synergistic oligodeoxynucleotide (iSN‐ODN, named iSN34) from Lactobacillus rhamnosus GG that has immunosynergistic activity upon costimulation of target cells with ligands of Toll‐like receptor 9 (TLR9). Here, we extend that observation by demonstrating the synergistic induction (in mouse splenocytes) of IL‐6 by the combination of iSN34 with cell wall components of bacteria and fungi. We observed that splenocytes pretreated with iSN34 and then costimulated with agonists for TLR1/2 (Pam₃CSK₄), TLR4 (lipopolysaccharide), or TLR2/6 (Zymosan) exhibited enhanced accumulation of IL‐6. These results suggested that the combination of iSN34 with TLR1/2, TLR4, or TLR2/6 agonists may permit the induction of a potent immune response.     

Screening and investigation Lactobacillius spp. to improve Secale cereale silage quality

Silage is a high‐moisture‐content forage that is used to feed livestock. Using silage as feed is economically feasible and suitable for cattle management. Secale cereale is considered as a high‐quality forage plant. After the heading stage the biomass of S. cereale increases by up to 30%; however, lignification in the cell wall causes low fermentation efficiency and coefficient digestibility, and it may also be contaminated by mycotoxin‐producing fungi and can impair the quality of the silage. Therefore, the aim of this study was to screen beneficial Lactobacillius spp. for S. cereale silage to reduce fungi contamination and to increase fibrinolytic ability in the silage. Up to 100 days fermentation period and total 180 lactic acid bacteria were isolated and only a single isolate, R4‐26, showed strong acidification ability and antifungal activities against mycotoxin‐producing fungus. According to quantitative RT‐PCR results, the mycotoxin‐producing fungal contamination was significantly reduced in the S. cereale silage containing the R4‐26 isolate. In enzymatic assays, only a single isolate, R48‐27 demonstrated cellulase, xylanase, chitinase and esterase activities. In conclusion, the beneficial Lactobacillius spp. strains have the ability to reduce mycotoxigenic fungi contamination and increase fibrinolytic activity to improve quality of the S. cereale silage.     

Plasma pharmacokinetics of quinocetone in ducks after oral and intravenous administration

Quinocetone (QCT), an antimicrobial growth promoter, is widely used in food‐producing animals. However, information about pharmacokinetics (PK) of QCT in ducks still remains unavailable up to now. In this study, QCT and its major metabolites (1‐desoxyquinocetone, di‐desoxyquinocetone and 3‐methyl‐quinoxaline‐2‐carboxylic) in ducks were studied using a simple and sensitive UHPLC‐MS/MS assay. Twenty ducks were divided into two groups. (n = 10/group). One group received QCT by oral administration at dose of 40 mg/kg while another group received QCT intravenously at 10 mg/kg. Plasma samples were collected at various time points from 0 to 96 hr. QCT and its major metabolites in duck plasma samples were extracted by 1 ml acetonitrile and detected by UHPLC‐MS/MS, with the gradient mobile phase that consisted of 0.1% formic acid in water (A) and acetonitrile (B). A noncompartment analysis was used to calculate the PK parameters. The results showed that following oral dosing, the peak plasma concentration (C_max) of QCT was 32.14 ng/ml and the area under the curve (AUCINF_obs) was 233.63 (h ng)/ ml. Following intravenous dosing, the C_max, AUCINF_obs and Vss_obs were 96.70 ng/ml, 152.34 (h ng)/ ml and 807.00 L/kg, respectively. These data indicated that the QCT was less absorbed in vivo following oral administration, with low bioavailability (38.43%). QCT and its major metabolites such as 1‐desoxyquinocetone and 3‐methyl‐quinoxaline‐2‐carboxylic were detected at individual time points in individual ducks, while the di‐desoxyquinocetone was not detected in all time points in all ducks. This study enriches basic scientific data about pharmacokinetics of QCT in ducks after oral and intravenous administration and will be beneficial for clinical application in ducks.     

Dispersal of invasive Phytolacca americana seeds by birds in an urban garden in China

Although seed dispersal is a key process determining the regeneration and spread of invasive plant populations, few studies have explicitly addressed the link between dispersal vector behavior and seedling recruitment to gain insight into the invasion process within an urban garden context. We evaluated the role of bird vectors in the dispersal of pokeweed (Phytolacca americana), a North American herb that is invasive in urban gardens in China. Fruiting P. americana attracted both generalist and specialist bird species that fed on and dispersed its seeds. The generalist species Pycnonotus sinensis and Urocissa erythrorhyncha were the most frequent dispersers. Seedling numbers of P. americana were strongly associated with the perching behavior of frugivorous birds. If newly recruited bird species use seedling‐safe perching sites, the P. americana will regenerate faster, which would enhance its invasive potential. Based on our observations, we conclude that the 2 main bird vectors, P. sinensis and U. erythrorhyncha, provide potential effective dispersal agents for P. americana. Our results highlight the role of native birds in seed dispersal of invasive plants in urban gardens.     

Conjunctival bacterial and fungal flora in healthy horses in the UK

Objectives To describe the bacterial and fungal flora of the normal conjunctiva of horses in the UK; to determine the effect of horse age, sex, geographic location, and housing on this flora; and to determine the most appropriate antimicrobial drug(s) for prophylactic treatment of corneal ulcers. Animal studied A total of 60 adult healthy horses were studied. Procedure Swabs of the conjunctiva were obtained from 60 horses housed in two locations within the UK. Specimens were cultured for aerobic bacteria and fungi, and sensitivity against six ophthalmic antimicrobials assessed. The effect of age, sex, location and housing on the frequency of microbial isolation was evaluated. Results Fifty‐four bacterial isolates, representing thirteen genera of bacteria were cultured from 31 (52%) horses. The most frequently isolated bacterial species was Acinetobacter sp (17/32 horses). The majority of isolates (28/54: 52%) were gram‐positive. Three genera of fungi (Mucor, Absidia and Aspergillus spp) were isolated from eight (13%) horses. There was no significant effect of geographic location, sex, age or housing on frequency of microbial isolation. Horses from which gram‐negative bacteria were isolated were significantly older than horses from which gram‐positive bacteria were isolated. High efficacy (greater than 90% of isolates sensitive in vitro) was displayed by chloramphenicol, gentamicin and tetracycline. Conclusions The microbial species isolated are comparable with studies performed in other countries, although the frequency of Acinetobacter isolation was higher which may reflect a geographic difference. The topical antimicrobials gentamicin and chloramphenicol are appropriate first line antimicrobials for empirical treatment of corneal ulcers in the UK.     

Ochratoxin A cytotoxicity on Madin–Darby canine kidney cells in the presence of alpha‐tocopherol: Effects on cell viability and tight junctions

Ochratoxin A (OTA) is a potent nephrotoxic fungi metabolite that affects animal and human health. At the cellular level, OTA is able to alter functions and viability by several mechanisms of action. Several strategies to counteract its toxicity have been studied. We investigated the role of α‐tocopherol in counteracting OTA oxidative damage in Madin–Darby canine kidney (MDCK) cells by pre‐incubating the cells for 3 hr with the antioxidant (1 nm , 10 μm ) and then adding OTA (0–1.2 μg/ml) for the following 24 hr. Cell viability, lactate dehydrogenase (LDH) release, TUNEL staining and occludin and Zo1 localization by immunofluorescence were determined. Here, 1 nm α‐tocopherol was shown to significantly reduce (p < .05) the cytotoxicity, LDH release and apoptotic rate induced by OTA. The presence of the antioxidant at the same concentration maintained the localization of occludin and Zo1 in the rim of the MDCK cells after the 24‐hr OTA exposure. These results indicate that a low concentration of α‐tocopherol could block OTA toxicity, supporting its defensive role in the cellular membrane.     

Seasonal effects on the aerobic bacterial and fungal conjunctival flora of normal thoroughbred brood mares in Florida

Objective To evaluate seasonal effects on the presence or absence of fungal and aerobic bacterial flora of the conjunctival fornix of normal Florida Thoroughbred horses. Sample population Both eyes of 100 horses. Procedure Horses with normal anterior segment ophthalmic examinations from three farms in north central Florida were included. Each animal had the ventral conjunctival fornix of each eye swabbed with sterile culturettes. Samples were taken in October, January, April, and July (1999–2000). Aerobic and fungal cultures were plated. Bacterial cultures were reviewed at 24 and 48 h. Fungal cultures were reviewed weekly for 4 weeks. Logistic regression analysis with season as a factor and age of the horse as a covariate was performed. Statistical significance was set at P < 0.01. Results Horses ranged from 3 to 24 years of age, with a median age of 9 years. Twenty‐four genera of bacteria and 35 genera of fungi were recovered. Corynebacterium sp., Staphylococcus sp., Bacillus sp. and Moraxella sp. were the bacteria most frequently isolated. Mold species, dematiaceous mold species, Chrysosporium sp., Cladosporium sp., and Aspergillus sp. were the most frequently recovered fungi. Season did not have a significant effect on the presence of microorganisms isolated for individual horses adjusted for age. Younger horses had an increased incidence of gram‐negative rods and fungal isolates. The number of bacteria and fungi isolated are not uniform across seasons. Conclusion There were no significant differences between the number or type of organisms cultured during the sampling seasons in normal Florida horses. A large range of normal bacterial and fungal flora were isolated from these horses. The number of bacteria and fungi isolated are not uniform across seasons. The likelihood of detecting an organism depends on the horses’ age.     

Seed traits and taxonomic relationships determine the occurrence of mutualisms versus seed predation in a tropical forest rodent and seed dispersal system

Although many studies have been carried out on plant–animal mutualistic assemblages, the roles of functional traits and taxonomy in determining both whether interactions involve mutualisms or predation and the structure of such assemblages are unclear. We used semi‐natural enclosures to quantitatively assess the interaction strengths between seeds of 8 sympatric tree species and 4 rodent species in a tropical forest in Xishuangbanna, Yunnan, Southwest China. We found 2 clusters of species in the seed–rodent network represented by 2 genera in the Fagaceae (Castanopsis, Lithocarpus). Compared to seeds of 3 Castanopsis species, seeds with heavy weight, hard coat or caloric content (including 3 Lithocarpus species) were eaten less and more frequently hoarded by rodents. In turn, hoarded seeds showed less predation and more mutualism with rodents. Our results suggest that seed traits significantly affected the hoarding behavior of rodents, and, consequently, the occurrence of mutualisms and predation as well as assemblage structure in the plant–animal seed dispersal system. Taxonomically‐related species with similar seed traits as functional groups belong to the same substructures in the assemblage. Our results indicate that both seed traits and taxonomic relationships may simplify thinking about seed dispersal systems by helping to elucidate whether interactions are likely to be dominated by predation or mutualism.     

Differentiation in seed hoarding among three sympatric rodent species in a warm temperate forest

Although seed hoarding by rodents has been extensively studied, differentiation in seed‐hoarding behaviors among sympatric rodent species has not been well investigated. Using semi‐natural enclosures, we demonstrated that three sympatric rodent species showed clear differentiation in food selection, scatter versus larder hoarding behaviors and eating behaviors when offered seeds of four plant species from a warm temperate forest in northern China. The large field mouse Apodemus peninsulae preferred seeds of wild apricot (Prunus armeniaca) and Liaodong oak (Quercus liaotungensis), whereas the Chinese white‐bellied rat Niviventor confucianus preferred seeds of cultivated walnut and Liaodong oak, and the David's rock squirrel Sciurotamias davidianus preferred seeds of cultivated walnut, wild apricot and Liaodong oak. All three rodents showed larder hoarding of seeds from all four plant species, but the large field mouse showed scatter hoarding of wild apricot, and the David's rock squirrel showed scatter hoarding of Liaodong oak and wild walnut. Acorns of Liaodong oak, which have a soft seed hull, were more often eaten in situ, whereas wild walnuts, which have a hard seed hull and more tannin, were less hoarded by all rodent species. Differentiation in the scatter versus larder hoarding behaviors of sympatric rodent species suggests that sympatric rodents play different roles in the regeneration of different sympatric plant species.     

Does scatter‐hoarding of seeds benefit cache owners or pilferers?

The scatter‐hoarding behavior of granivorous rodents plays an important role in seed dispersal and seedling regeneration of trees, as well as the evolution of several well‐known mutualisms between trees and rodents in forest ecosystems. Because it is difficult to identify seed hoarders and pilferers under field conditions by traditional methods, the full costs incurred and benefits accrued by scatter‐hoarding have not been fully evaluated in most systems. By using infrared radiation camera tracking and seed tagging, we investigated the benefits and losses of scatter‐hoarded seeds (Camellia oleifera) for 3 sympatric rodent species (Apodemus draco, Niviventer confucianus and Leopoldamys edwardsi) in a subtropical forest of Southwest China during 2013 to 2015. We established the relationships between the rodents and the seeds at the individual level. For each rodent species, we calculated the cache recovery rate of cache owners, as well as conspecific and interspecific pilferage rates. We found that all 3 sympatric rodent species had a cache recovery advantage with rates that far exceeded average pilferage rates over a 30‐day tracking period. The smallest species (A. draco) showed the highest rate of scatter‐hoarding and the highest recovery advantage compared with the other 2 larger species (N. confucianus and L. edwardsi). Our results suggest that scatter‐hoarding benefits cache owners in food competition, supporting the pilferage avoidance hypothesis. Therefore, scatter‐hoarding behavior should be favored by natural selection, and plays a significant role in species coexistence of rodent community and in the formation of mutualism between seeds and rodents in forest ecosystems.     

Effects of insect infestation on rodent‐mediated dispersal of Quercus aliena: results from field and enclosure experiments

Rodents influence plant establishment and regeneration by functioning as both seed predators and dispersers. However, these rodent–plant interactions can vary significantly due to various environmental conditions and the activity of other insect seed predators. Here, we use a combination of both field and enclosure (i.e. individual cage and semi‐natural enclosure) experiments, to determine whether rodents can distinguish sound seeds from those infested with insects. We also demonstrate how such responses to insects are influenced by food abundance and other environmental factors. We presented rodents with 2 kinds of Quercus aliena seeds (sound and insect‐infested seeds) in a subtropical forest in the Qinling Mountains, central China, from September to November of 2011 to 2013. The results showed that rodents preferred to hoard and eat sound seeds than infested seeds in the field and semi‐natural enclosure, while they preferred to eat infested seeds over sound seeds in the individual cages. In addition, both hoarding and eating decisions were influenced by food abundance. Rodents hoarded more sound seeds in years of high food abundance while they consumed more acorns in years of food shortage. Compared with field results, rodents reduced scatter‐hoarding behavior in semi‐natural enclosures and ate more insect‐infested seeds in smaller individual cages. These results further confirm that rodents distinguish infested seeds from non‐infested seeds but demonstrate that this behavior varies with conditions (i.e. environment and food abundance). We suggest that such interactions will influence the dispersal and natural regeneration of seeds as well as predation rates on insect larvae.     

β‐hydroxy‐β‐methyl butyrate promotes leucine metabolism and improves muscle fibre composition in growing pigs

The aim of this study was to investigate the effects of excess leucine (Leu) vs. its metabolites α‐ketoisocaproate (KIC) and β‐hydroxy‐β‐methyl butyrate (HMB) on Leu metabolism, muscle fibre composition and muscle growth in growing pigs. Thirty‐two pigs with a similar initial weight (9.55 ± 0.19 kg) were fed 1 of 4 diets for 45 days: basal diet, basal diet + 1.25% L‐Leu, basal diet + 1.25% KIC‐Ca, basal diet + 0.62% HMB‐Ca. Results indicated that relative to the basal diet and HMB groups, Leu and KIC groups exhibited increased Leu concentrations and decreased concentrations of isoleucine, valine and EAAs in selected muscle (p < 0.05) and had lower mRNA levels of MyHC I and higher expression of MyHC IIx/IIb (p < 0.05), and there was no significant difference between the basal and HMB‐supplemented groups. Moreover, the mRNA expression levels of AMPKα and UCP3 were higher but the myostatin mRNA levels were lower in the soleus muscle of the HMB group than those from other groups (p < 0.05). These findings demonstrated that doubling dietary Leu content exerted growth‐depressing effects in growing pigs; dietary KIC supplementation induced muscular branched‐chain amino acid imbalance and promoted muscle toward a more glycolytic phenotype; while dietary HMB supplementation promoted the generation of more oxidative muscle types and increased muscle growth specially in oxidative skeletal muscle, and these effects of HMB might be associated with the AMPKα‐Sirt1‐PGC‐1α axis and mitochondrial biogenesis.     

Expression of claudin‐5 in hepatoid gland biopsies

Claudins are integral membrane proteins involved in the structure of the tight junctions found in epithelial and endothelial cells. This study evaluated the expression of claudin‐5 in 67 hyperplastic and neoplastic lesions of canine hepatoid glands. These included normal hepatoid glands (n = 10), nodular hyperplasia (n = 10), adenomas (n = 12), epitheliomas (n = 15), differentiated carcinomas (n = 15) and anaplastic carcinomas (n = 15). There was intense lateral membrane expression of claudin‐5 on epithelial cells from normal hepatoid glands, nodular hyperplasia and adenomas, but expression was weaker in hepatoid gland epitheliomas. Basal reserve cells from normal glands, nodular hyperplasia, adenomas and epitheliomas never expressed claudin‐5. There was membrane‐bound immunoreactivity for claudin‐5 in selected areas of the epitheliomas where the cells exhibited typical hepatoid features. The weak expression of claudin‐5 molecule in epitheliomas may nevertheless lead to cellular disorientation, detachment and invasion. Claudin‐5 expression seemed to be helpful in distinguishing poorly differentiated carcinomas, differentiated carcinomas and epitheliomas of the hepatoid glands. Increased claudin‐5 expression by invasive anaplastic carcinomas may facilitate invasion and metastasis through the activation of matrix metalloproteinases.     

Cytologic identification of fungal arthritis in a Labrador Retriever with disseminated Talaromyces helicus infection

An 8‐year‐old, neutered male Labrador Retriever presented with acute forelimb lameness. Clinical signs progressed over one week. On physical examination, right cubital joint effusion and bilateral axillary lymphadenomegaly were noted, and severe internal lymphadenomegaly was observed ultrasonographically. Granulomatous lymphadenitis with intralesional fungi was noted cytologically, and the dog was ultimately diagnosed with disseminated Talaromyces helicus infection via PCR of a pure isolate. Extensive medical therapy was pursued, and months later, an arthrocentesis was performed due to continued lameness and severe cubital joint effusion. The synovial fluid contained increased numbers of neutrophils, macrophages, and multinucleated giant cells. Frequent fungal hyphae were found both intracellularly and extracellularly. These basophilic organisms were 2‐4 µm in width with internal eosinophilic granules, roughly parallel walls, and occasional to frequent septa. Round to oval yeast‐like forms with thin, clear halos were also occasionally identified. Due to the severity of clinical signs, the right thoracic limb was amputated. Histologic examination of the cubital joint revealed marked granulomatous synovitis, fasciitis, panniculitis, and osteomyelitis, all with intralesional fungi. Talaromyces helicus is a very rare cause of disease, reported only in one other dog. Granulomatous lymphadenitis appears to be a feature of this disease, but this report is the first to describe a significant synovial component.