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Importance: Porcine epidemic diarrhea virus (PEDV) binds to particular cell surface receptors to penetrate cells. The virus specifically identifies certain carbohydrate structures present on the surface of the cell to facilitate the binding process. Nevertheless, the influence of viral infections on specific alterations of glycoconjugates in the small intestines remains unexplored., Objective: This work aimed to examine the alterations in glycoconjugates in the small intestines of piglets naturally infected with PEDV using lectin histochemistry., Methods: Six piglets including three PEDV-infected and three non-infected piglets were evaluated. Small intestinal samples were histopathologically examined, and lectin histochemistry was performed., Results: Piglets infected with PEDV had significant histological abnormalities in their small intestines, such as pronounced villous atrophy, varying degrees of villous fusion, and diverse mucosal alterations. Specific regions of the duodenum, jejunum, and ileum showed discernible variations in glycoconjugate distribution, as determined by lectin histochemistry. Compared with the controls, the PEDV-infected piglets showed significant changes in N-acetylglucosamine- and galactose-binding lectins (particularly wheat germ agglutinin and Arachis hypogaea (peanut) agglutinin) in multiple intestinal regions., Conclusions and Relevance: These findings can enhance understanding of how viruses such as PEDV impact the glycoconjugate composition of the small intestines and emphasize the potential connection between the pathogenesis of PEDV and glycoconjugate., Competing Interests: The authors declare no conflicts of interest., (© 2024 The Korean Society of Veterinary Science.)

The African straw-coloured fruit Bat, Eidolon helvum, is a Yinpterochiroptera bat that is of public health and ecological importance. This study investigated the light microscopic morphology, morphometry and histochemistry of the skin and its glands across the cheek, withers, croup, ventral abdomen, perianal region, wing membrane, intercrural membrane and foot pad of the species. Sweat glands were absent and hypodermis was observed only in the withers, croup and ventral abdomen. The croup had the highest density of hair follicles while the perianal region had the highest density of sebaceous gland acini. The thicknesses of the dermis and epidermis were significantly high in the withers and foot pad respectively. Hair follicles and sebaceous glands were absent in the wing membrane. The sebaceous glands had melanin pigmentation at the periphery and stained positive to Alcian Blue-periodic acid Schiff at the withers and ventral abdomen. Epidermal and dermal thicknesses reported in this study were generally higher than those of Yangochiroptera bats and the presence of sebaceous glands and hair follicles in the wing membrane were variable in different bat species. This study has provided baseline information on the light microscopic morphology and morphometry of the skin and its glands across the various body regions in the African straw-coloured fruit bat. This information would be helpful in the understanding, handling, skin disease treatment and management of the bat in captivity for biomedical research purposes., (© 2023 Wiley-VCH GmbH. Published by John Wiley & Sons Ltd.)

Background: Distinguishing primary and secondary pulmonary neoplasms can be challenging via cytology, and a rapid, inexpensive diagnostic tool to differentiate these neoplasms is unavailable. Alkaline phosphatase cytochemistry (ALP-CC) has been used to identify primary pulmonary carcinomas in human patients, and we hypothesized it could be applied to canine lung aspirates., Objective: We aimed to characterize ALP-CC expression in fine-needle aspirate (FNA) samples of canine pulmonary neoplastic and non-neoplastic tumors., Methods: A retrospective case search was conducted to identify cases with contemporaneous cytology and histopathology reports from pulmonary lesions, including neoplastic and non-neoplastic etiologies. Slides prepared from pulmonary aspirates were stained for ALP-CC activity, and the percentage of ALP-CC-positive primary pulmonary epithelial tumors was determined. To characterize the ALP-CC expression in non-neoplastic cellular constituents of pulmonary FNA samples, mesothelial cells were also evaluated., Results: Forty-eight canine cases met the inclusion criteria. ALP-CC-positive cells were seen in both neoplastic and non-neoplastic lesions. In non-neoplastic lesions, pulmonary epithelial cells were ALP-CC positive. Eighty-nine percent of primary pulmonary epithelial neoplasms were ALP-CC positive, and no ALP-CC positivity was noted in mesothelial cells. ALP-CC-positive neoplastic cells were seen in a metastatic amelanotic melanoma., Conclusions: Primary pulmonary epithelial neoplasms are frequently ALP-CC positive, but such positivity is not restricted to this tumor type. Non-neoplastic pulmonary epithelial cells can be ALP-CC positive, whereas mesothelial cells are negative., (© 2023 The Authors. Veterinary Clinical Pathology published by Wiley Periodicals LLC on behalf of American Society for Veterinary Clinical Pathology.)

This study describes the morphology and histochemistry of the digestive tube of the herbivorous fish Rhytiodus microlepis, which is commonly found in the Amazonian floodplain lakes, Brazil. Thirty-eight fish were used in this study. The digestive tube was measured and dissected for anatomical description, and stained with (i) haematoxylin and eosin, for histological analyses; (ii) periodic acid-Schiff, for detected neutral mucins; and (iii) Alcian Blue (AB) pH 1.0 and 2.5 for acid and sulphated acid mucins. The oesophagus, with its secretory cells and taste buds, is adapted for lubrication of vegetable matter, resistance to friction and reception of stimuli. The stomach is U-shaped and rich in gastric glands, particularly in cardiac and fundic regions, which are adapted to receive large amounts of food, and promotes the digestion. The intestine comprises more than 70% of the digestive tube. All portions of the intestine, except the rectum, have the same pattern of mucosal folds, and the goblet cells reacted positively to all histochemical methods (PAS, AB 1.0 and 2.5), while rodlet cells reacted only to periodic acid-Schiff (PAS) staining. Neutral mucins and sulphated acid mucins predominated in the anterior portion of the intestine and a high concentration of carboxylated acid mucosubstances were present in the other portions. The rectum showed a strong reaction to all histochemical methods. The muscular layer of the intestine consists of three layers, which showed features presumably related to the rigid nature of the food. A simple squamous epithelium constitutes the serous layer over the entire length of the tube., (© 2021 Wiley-VCH GmbH.)

Distribution of tripeptidyl peptidase I (TPPI) activity in the structures of porcine lumbar spinal ganglia (LSG) was studied by enzyme histochemistry on cryostat sections from all the ganglia using the substrate glycyl-L-prolyl-L-methionyl-5-chloro-1-anthraquinonyl hydrazide (GPM-CAH) and 4-nitrobenzaldehyde (NBA) as visualization factor. Light microscopic observations showed TPPI activity in almost all the LSG structures. The enzyme reaction in different cell types was compared semi-quantitatively. Strong reaction was observed in the small neurons, satellite ganglia cells and some nerve fibers. Weak reactivity was found in the large sensory somatic neurons, whereas moderate reaction for TPPI was determined in the middle sensory somatic neurons and some nerve fibers. Statistical analysis by one-way ANOVA showed no significance of difference (when p⟨0.05) for the number of TPPI positive neurons per mm2. The original data obtained by the enzyme histochemistry method give us a reason to presume that TPPI actively participates in the functions of all the neuronal structures in porcine LSG. According to our results, it could be suggested that TPPI activity is important for the functions of autonomic and somatic sensory neurons., (Copyright© by the Polish Academy of Sciences.)

The health effects of mercury vapor exposure on the brain in volcanic areas have not been previously addressed in the literature. However, 10% of the worldwide population inhabits in the vicinity of an active volcano, which are natural sources of elemental mercury emission. To evaluate the presence of mercury compounds in the brain after chronic exposure to volcanogenic mercury vapor, a histochemical study, using autometallographic silver, was carried out to compare the brain of mice chronically exposed to an active volcanic environment (Furnas village, Azores, Portugal) with those not exposed (Rabo de Peixe village, Azores, Portugal). Results demonstrated several mercury deposits in blood vessels, white matter and some cells of the hippocampus in the brain of chronically exposed mice. Our results highlight that chronic exposure to an active volcanic environment results in brain mercury accumulation, raising an alert regarding potential human health risks. These findings support the hypothesis that mercury exposure can be a risk factor in causing neurodegenerative diseases in the inhabitants of volcanically active areas.

The development of the epidermis of octopus, Octopus vulgaris, throughout its life cycle was studied by conventional staining and histochemical techniques using lectins. The mantle, the arm and the two parts of the suckers: the infundibulum and the acetabulum were analysed independently. With the exception of the suckers, the general morphology of the epidermis does not vary from the first days post-hatching to adulthood. In general terms, histochemical techniques do not indicate changes in the composition of glycoconjugates of the epidermis main cells, epithelial and secretory cells. The epithelial cells of the mantle and arm show positivity for mannose (ConA+) in their apical portions, indicating the presence of n-glycoproteins that, among other things, provide lubrication to the surface of the body. In the suckers, the apical surface of the infundibulum contains sulphated glycosaminoglycans of the N-acetylglucosamine type that provide adhesive properties. In addition to observing three types of mucocytes, m1 and m2 are characteristic of the mantle and arm, and m3 is found in the suckers. The paralarva epidermis is characterised by the presence of Kölliker's organs whose exact function is unknown. In this study, the absence of staining with alcian blue/periodic acid-Schiff(AB/PAS) prevents the possibility of attributing a secretory function. Nevertheless, the linkage of three lectins (WGA, LEL and GSL-I) in the fascicle of the organ suggests the presence of proteoglycans rich in N-acetylglucosamine that would mainly have a structural role., (© 2020 Blackwell Verlag GmbH.)

Enterospora nucleophila is a microsporidian responsible for an emaciative disease in gilthead sea bream ( Sparus aurata ). Its intranuclear development and the lack of in vitro and in vivo models hinder its research. This study investigated the associated lesions, its detection by quantitative polymerase chain reaction, and the cellular immune response of naturally infected fish. The intensity of infection in the intestine was correlated with stunted growth and reduced body condition. At the beginning of the outbreaks, infection prevalence was highest in intestine and stomach, and in subsequent months, the prevalence decreased in the intestine and increased in hematopoietic organs and stomach. In heavy infections, the intestine had histologic lesions of enterocyte hypercellularity and proliferation of rodlet cells. Infected enterocytes had E. nucleophila spores in the cytoplasm, and a pyknotic nucleus, karyorhexis or karyolysis. Lymphocytes were present at the base of the mucosa, and eosinophilic granule cells were located between the enterocytes. In intestinal submucosa, macrophage aggregates containing spores were surrounded by lymphocytes and granulocytes, with submucosal infiltration of granulocytes. Macrophage aggregates appeared to develop into granulomata with necrotic areas containing parasite remnants. Immunohistochemistry revealed mast cells as the main type of granulocyte involved. Abundant IgM + and IgT + cells were identified by in situ hybridization in the submucosa when intracytoplasmic stages were present. This study describes the lesions of E. nucleophila in gilthead sea bream, an important aquaculture species.

A 4-month-old female mixed-breed cat showed gait disturbance and eventual dysstasia with intention tremor and died at 14 months of age. Postmortem histological analysis revealed degeneration of neuronal cells, alveolar epithelial cells, hepatocytes, and renal tubular epithelial cells. Infiltration of macrophages was observed in the nervous system and visceral organs. The cytoplasm of neuronal cells was filled with Luxol fast blue (LFB)-negative and periodic acid-Schiff (PAS)-negative granules, and the cytoplasm of macrophages was LFB-positive and PAS-negative. Ultrastructurally, concentric deposits were observed in the brain and visceral organs. Genetic and biochemical analysis revealed a nonsense mutation (c.1017G>A) in the SMPD1 gene, a decrease of SMPD1 mRNA expression, and reduced acid sphingomyelinase immunoreactivity. Therefore, this cat was diagnosed as having Niemann-Pick disease with a mutation in the SMPD1 gene, a syndrome analogous to human Niemann-Pick disease type A.

The purpose of the present study was to determine the histological and ultrastructural changes in the luminal epithelium of the shell gland associated with natural moulting. Samples of the shell gland from laying (32 weeks old) and moulting (75 weeks old) hens were studied using histological, histochemical and electron microscopic techniques. In addition, TUNEL was used to demonstrate the distribution of apoptotic cells in the luminal epithelium of the shell gland. Autophagy, characterized by the presence of autophagosomes and autolysosomes, was evident in the early stages of degeneration in non-ciliated, ciliated and mitochondrial cells. The intermediate and advanced stages of regression in non-ciliated as well as mitochondrial cells occurred via apoptosis, while both apoptotic and necrotic ciliated cells were observed during the later stages of degeneration. The results of the present study suggest that a synergy of autophagy, apoptosis and necrosis is involved in the involution of the shell gland during natural moulting., (© 2020 Blackwell Verlag GmbH.)

Infection of small ruminants with peste des petits ruminants virus (PPRV) and goatpox virus (GTPV) are endemic and can have devastating economic consequences in Asia and Africa. Co-infection with these viruses have recently been reported in goats and sheep in Nigeria. In this study, we evaluated samples from the lips of a red Sokoto goat, and describe co-infection of keratinocytes with PPRV and GTPV using histopathology and transmission electron microscopy. Eosinophilic cytoplasmic inclusion bodies were identified histologically, and ultrastructural analysis revealed numerous large cytoplasmic viral factories containing poxvirus particles and varying sizes of smaller cytoplasmic inclusions composed of PPRV nucleocapsids. These histopathological and ultrastructural findings show concurrent infection with the 2 viruses for the first time as well as the detection of PPRV particles in epithelial cells of the mucocutaneous junction of the lip.

Background: Free-living cetaceans are exposed to a wide variety of stressful situations, including live stranding and interaction with human beings (capture myopathy), vessel strikes, and fishing activities (bycatch), which affect their wellbeing and potentially lead to stress cardiomyopathy (SCMP)., Methods: Here, the authors aimed to characterise SCMP of stranded cetaceans as an injury resulting from extreme stress responses, based on pathological analyses (histological, histochemical and immunohistochemical). Specifically, the authors examined heart samples from 67 cetaceans found ashore (48 live strandings, seven dead from ship collision and 12 dead from bycatch) on the coast of Spain, more specifically in the Canary Islands from 2000 to 2016 and Andalusia from 2011 to 2014., Results: The microscopic findings were characterised by vascular changes, acute or subacute cardiac degenerative necrotic lesions, interstitial myoglobin globules, and infiltration of inflammatory cells. Immunohistochemically, cardiac troponin I, cardiac troponin C and myoglobin were depleted, along with fibrinogen being expressed in the degenerated/necrotic cardiomyocytes. A perivascular pattern was also identified and described in the damaged cardiomyocytes., Conclusions: This study advances current knowledge about the pathologies of cetaceans and their implications on conserving this group of animals by reducing mortality and enhancing their treatment and subsequent rehabilitation to the marine environment., Competing Interests: Competing interests: None declared., (© British Veterinary Association 2019. No commercial re-use. See rights and permissions. Published by BMJ.)

The changes in the ovarian stroma of the fish during their spawning season become it an excellent biological model for studies on cellular and vascular elements due to the intense tissue remodeling in fish occur naturally throughout this critical period. The present study aims to investigate the cellular and vascular components of the ovarian stroma of Redbelly tilapia during the spawning season by conventional, immunohistochemical stains as well as to detect the ultrastructural characteristics for each stromal component. The histological examinations revealed a series of blood vessels with special structures, include throttle artery, glomus, spirally oriented arterioles, modified arteries, and veins as well as arteriovenous anastomosis. Various types of cells were detected in the stroma include; telocytes, rodlet cells, mast cells, eosinophils, neutrophils, lymphocytes, fibroblasts, macrophages, melanocytes, adipocytes, dendritic cells, and endocrine (steroidogenic, interstitial) cells. Moreover, these stromal cells showed a broad range of staining affinity against c-kit, desmin, and s100-protein. Bundles of nerve fibers were detected between the follicles. This study exposed various cellular and vascular components with distinct functions in the ovary of Redbelly tilapia during the spawning season., (Copyright © 2019 Elsevier Ltd. All rights reserved.)

The health of oviduct circulation is very important for normal egg production. The present work describes the histology and histochemistry of vessels of different types on the oviduct of quail. Arteria, venae and lymphatics of the oviduct were described clearly. In addition, special vascular elements were observed presenting circular, longitudinal and oblique smooth muscle fibres on the blood vessel wall. Glomus cells were observed at typical glomus vessels, atypical glomus vessels and on the wall of some of arteria and arteriolae. Also, spirally oriented blood vessels were observed. Dark brown pigment was detected in the wall of arteria, venae, lymphatics and the special types of blood vessels., (© 2019 Blackwell Verlag GmbH.)

The tortuous nature of the ovine cervix restricts the transcervical passage of the cannula, and many studies have aimed to understand the endocrine mechanism of the remodelling of cervical tissue in adult ewe. However, little is known about the remodelling of the cervical tissue during the prepubertal development of the lambs. To obtain histochemical and biochemical evidence about the nature of the prepubertal development of the cervix of the ewe, cervices of Corriedale lambs obtained at 0, 1, 2, 4, 6 and 8 months of age (n = 5 to 6 in each) were processed. Neutral and acidic glycosaminoglycans (by PAS-Alcian stain) were weakly in the cervical stroma and not shown change during the development, whereas the percentage volume of fibrillar collagen (by van Gieson stain) increases throughout the experimental period in the superficial fold stroma and deep wall stroma (p < 0.05). The relative cervical weight (g/kg of body weight) and the collagen concentration (by spectrophotometry, mg/mg wet tissue) showed an early decreasing phase from months 0 to 4 and a later increasing phase from months 4 to 8 (p < 0.05). The latent form of matrix metalloproteinase-2 (MMP-2) detected by gelatin zymography (ng/mg protein) decreased from months 0 to 2 and increased from months 4 to 8, whereas the activated form decreased from months 0 to 2, remained low until month 6 and then recovered on month 8 (p < 0.0001). Data suggest that the relative cervical weight biphasic pattern during the development is related to MMP-2-dependent changes in the collagen content., (© 2019 Blackwell Verlag GmbH.)

The present study describes histochemical and immunohistochemical characteristics of the spiral valve and its associated lymphoid tissue (GALT) in the dogfish Scyliorhinus canicula. The mucosal surface of the spiral valve represents the first line of defense against pathogens coming from the external environment through food. Epithelial, mucus and immune cells play a key role in controlling the inflammatory response. Valve intestine of S. canicula had many folds lined by simple columnar cells and goblet cells, which later reacted positive to PAS, AB and AB-PAS, histochemical stains differentiated the different types of mucins; lectin histochemistry (PNA and WGA), detected neutral and acid mucins secreted that plays an important role in protection against invading pathogens. Integrin α5β1 was expressed in enterocytes that line the valve's folds with greater marking in the apical part of the cells. Laminin was found on the apical side of the epithelium, in fibrillar and cellular elements of the lamina propria and in the muscularis mucosa. In the spiral valve gut-associated lymphoid tissue (GALT) has been studied. For the first time, massive leucocytes aggregates were identified by confocal immunofluorescence techniques, using the following antibodies: TLR2, S100, Langerin/CD207. Our results expand knowledge about Dogfish valve intestine giving important news in understanding comparative immunology., (Copyright © 2019 Elsevier Ltd. All rights reserved.)

Fish are the most diverse species of all vertebrate groups, and their blood cells have shown variable characteristics in terms of morphology. Cytochemical staining for enzyme activity in blood leukocytes will help assess the immune function of fish. We characterize blood cells from crucian carp (Carassius auratus) and grass carp (Ctenopharyngodon idellus) by using a Diff-Quick stain as well as different cytochemical methods. Blood specimens obtained from crucian carp and grass carp were evaluated after cytochemical staining for acid phosphatase (ACP), alkaline phosphatase (ALP), naphthol AS chloroacetate esterase (AS-DNCE), naphthyl acetate esterase (NAE), α-naphthyl butyrate esterase (NBE), peroxidase (MPO) and periodic acid-Schiff's reaction (PAS) using commercial kits. Blood cell types were evaluated based on their morphological characteristics and the presence or absence of specific chromogen. The expression pattern of enzymes was similar between the two Cyprinidae and was also broadly consistent with other fish species. However, there were some interesting differences detected between crucian carp and grass carp, including naphthol AS chloroacetate esterase activity in monocytes, peroxidase activity and location in thrombocytes. The ACP, ALP and MPO expressions of different leukocytes of the two Cyprinidae were evaluated by Image Pro Plus and were analysed for statistical significant differences. This investigation provides basic haematology and enzyme activity analyses for crucian carp and grass carp and serves as an approach to evaluating the immune response of fish., (© 2018 Blackwell Verlag GmbH.)

Acute myeloid leukaemia (AML) is an uncommon, rapidly progressive neoplasm in dogs. The aim of this retrospective study was to characterize the clinical presentation, haematologic findings, diagnostic imaging results, treatment and survival time of a contemporary cohort of dogs with AML. Diagnosis was based on >20% blasts in bone marrow or blood identified as myeloid based on morphologic findings, flow cytometric immunophenotyping and cytochemical staining. Medical records of 35 dogs diagnosed with AML from 2007 to 2015 were included. Most dogs presented with inappetence (66%) and lethargy (57%) and physical examination findings of peripheral lymphadenopathy (74%) and tachypnea (62%). Common haematologic findings were quantifiable circulating blasts (85%; median blast count 35 700/μL; range: 300-276 500/μL), anaemia (median haematocrit 34%; range: 11%-52%) and thrombocytopenia (median 57 000/μL; range: 9000-252 000/μL). Bicytopenia and pancytopenia were each found in 44% of dogs. Follow-up information was available for 34 dogs. The overall median survival time from diagnosis was 19 days (range: 1-121 days). Clinical progression in some dogs was not as rapid as previously reported. Haematologic responses to various chemotherapeutics were documented in 3 dogs, with associated survival times of 62, 103 and 121 days. Dogs treated with prednisone or a combination of chemotherapy and prednisone had improved survival compared to dogs that received symptomatic care only (P < .0001). Our results show canine AML has an overlapping clinical presentation with lymphoma. The prognosis for canine AML remains extremely guarded. Further studies are needed to optimize therapeutic regimens for dogs with AML., (© 2017 John Wiley & Sons Ltd.)

Histochemical organization of the Caudata olfactory system remains largely unknown, despite this amphibian order showing phylogenetic diversity in the development of the vomeronasal organ and its primary centre, the accessory olfactory bulb. Here, we investigated the glycoconjugate distribution in the olfactory bulb of a semi-aquatic salamander, the Japanese sword-tailed newt (Cynops ensicauda), by histochemical analysis of the lectins that were present. Eleven lectins showed a specific binding to the olfactory and vomeronasal nerves as well as to the olfactory glomeruli. Among them, succinylated wheat germ agglutinin (s-WGA), soya bean agglutinin (SBA), Bandeiraea simplicifolia lectin-I (BSL-I) and peanut agglutinin showed significantly different bindings to glomeruli between the main and accessory olfactory bulbs. We also found that s-WGA, SBA, BSL-I and Pisum sativum agglutinin preferentially bound to a rostral cluster of glomeruli in the main olfactory bulb. This finding suggests the presence of a functional subset of primary projections to the main olfactory system. Our results therefore demonstrated a region-specific glycoconjugate expression in the olfactory bulb of C. ensicauda, which would be related to a functional segregation of the olfactory system., (© 2017 Blackwell Verlag GmbH.)

Ten apparently healthy, adult laughing doves were used to document detailed histological, histochemical and surface ultrastructural features of the nasal cavity and to investigate the structure-function relationship of the nasal cavity in this species. We observed that the nasal cavity of the laughing dove was composed of three main regions: nasal vestibule, respiratory and olfactory. Each region presented a characteristic epithelial lining. The epithelium varied along the nasal vestibule from keratinized stratified squamous rostrally to non-keratinized stratified squamous in the middle and stratified cuboidal in the caudal region of the nasal vestibule. The respiratory region was lined with pseudostratified columnar epithelium and was initially devoid of both goblet cells and cilia, but cilia then appeared and increased gradually in number close to the olfactory region. The caudal part of the respiratory region presented a stratified cuboidal epithelium. Strong alcianophilic, intra-epithelial mucous glands were identified, starting at the caudal region of the nasal vestibule and extended into the respiratory region. The olfactory region was lined with a pseudostratified epithelium that consisted of three different cell types: olfactory, support cells and basal cells. In conclusion, the current investigation presents new information concerning the histological, histochemical and ultrastructural features of the laughing dove's nasal cavity. Furthermore, the findings of this study may prove to be a valuable contribution to the avian histology and pathology literature., (© 2017 Blackwell Verlag GmbH.)

The increasing prevalence of yellow-bellied sliders (Trachemys scripta scripta) as pets in the European Union and also its utilization as animal models for experimental purposes makes crucial an accurate classification of their blood cells. The aim of this work was to provide a morphologic classification based on the cytochemical characteristics of the blood cells of 15 yellow-bellied sliders. Cytochemical stains included benzidine peroxidase, chloroacetate esterase, alpha-naphthyl butyrate esterase (with and without sodium fluoride), acid phosphatase (with and without tartaric acid), Sudan black B, periodic acid-Schiff and toluidine blue. Nuclear and cellular dimensions were also measured based on quick Romanowsky-type stained smears. Besides erythrocytes and thrombocytes, five types of white blood cells were identified: heterophils, eosinophils, basophils, lymphocytes and monocytes. The cytochemical patterns of heterophils, eosinophils and basophils were unique compared to those described for other chelonians. This paper provides a useful guideline for clinical settings and further haematological studies of this species., (© 2017 Blackwell Verlag GmbH.)

The presence of lesions on the pig carcass is an indicator of poor animal welfare and has economic impact as it downgrades the carcass value. The assessment of the age of lesions on the carcass may help identify risk factors and ultimately prevent their occurrence. The aim of this study was to assess the age of lesions on pig carcasses through spectrophotometric color evaluation and to relate the results with gene expression and histological and histochemical parameters. A total of 96 barrows were mixed 4 times over 3 d before slaughter and 80 lesions were selected after skin lesion observations to define 4 age categories: < 7 h (T1), 7-25 h (T2), 25-30 h (T3), and 49-54 h (T4). A nonlesioned skin area was used as a control. At slaughter, 3 biopsies per lesion and control skin were taken immediately after bleeding for analyses of gene expression (, , , , , , , , , ), skin histological characteristics (inflammation, erosion or ulceration, and necrosis), and enzyme activity (alkaline phosphatase and adenosine triphosphatase). The number of lesions was counted on each carcass, and the color was assessed visually by a pictorial chart and instrumentally through a spectrophotometer. Delta values (Δ) were calculated as the difference between the value of the lesion and the value of the control for all measures, except for the histological analysis. Results indicated that visual color observation was not sufficiently accurate to discriminate lesions by time of infliction ( > 0.10), while the spectrophotometer ΔL* and Δa* values variation allowed the identification of < 7 h or > 25 h old lesions ( < 0.05). Similarly, the expression of , , , , and genes was higher ( < 0.05) in < 7 h old lesions, while gene expression was higher ( < 0.05) in < 25 h old lesions. As for the histological analysis, the severity of inflammation was correlated with the age of the lesion (lower score in < 7 h old lesions and higher score in > 25 h old lesions; < 0.05). To conclude, the spectrophotometric color assessment of the carcass lesions at slaughter appears to be a reliable method to discriminate between fresh and older lesions on the carcass at the abattoir.

Background: Cell blocks are widely used in human cytopathology. Several techniques have been proposed to convert fluid specimens into solid material, which after embedding in paraffin can be used for histochemistry, immunohistochemistry, or molecular testing. In contrast, only in the last few years, have cell blocks begun to be used in the veterinary field., Objectives: The purpose of the study was to present the production and features of cell tube blocks (CTB) with veterinary liquid samples., Methods: Liquid samples including cerebrospinal fluid, cutaneous cyst fluid, pericardial and pleural effusions, bronchoalveolar lavage, urine, and blood were centrifuged in a microhematocrit centrifuge. Capillary tubes were broken at the liquid-solid interface and fixed in 10% formalin for 24 hours. After paraffin embedding, sections of CTB were used for different stains including immunohistochemistry., Results: The morphology and cellular detail in CTB sections were comparable to conventional histologic sections and other existing cell block techniques. The use of special stains such as Gram, Giemsa, alcian blue, and periodic acid-Schiff was straightforward, and immunohistochemistry results with antibodies to pan-cytokeratin, PAX-5, and CD3 were considered good., Conclusions: The CTB method was easily implementable under practice conditions (up to the fixation of the microhemtocrit tube), analogous to surgical biopsy submission for histology. Cell tube blocks can increase diagnostic accuracy when the technique is used in tandem after the cytologic evaluation, and the technique allows storage of fluids. Other advantages of CTB were the simplicity, low cost, and separation of erythrocytes from the nucleated cells, which was helpful in hemodiluted samples., (© 2017 American Society for Veterinary Clinical Pathology.)

The ultrastructural and cytochemical properties of peripheral blood cells of Gymnocypris eckloni were investigated by transmission electron microscopy and a range of cytochemical techniques to provide clear insight into the structure and function of blood cells from this fish. Ultrastructurally, erythrocytes, leucocytes (neutrophils, eosinophils, lymphocytes, monocytes), thrombocytes and plasma cells were identified in the peripheral blood of G. eckloni. The most special ultrastructural characteristics of blood cells in this fish were that neutrophils exhibited only one type of cytoplasmic granules containing an eccentric, spherical or oval electron-dense core, and eosinophils presented two types of granules with non-uniform electronic density and without crystalloids in their cytoplasm. Neutrophils, eosinophils, lymphocytes, monocytes and thrombocytes were positive for periodic acid-Schiff and α-naphthyl acetate esterase staining. Intense peroxidase positive staining was observed in neutrophils and monocytes, but not in eosinophils, lymphocytes and thrombocytes. Neutrophils, eosinophils and monocytes were stained positively for acid phosphatase, whereas lymphocytes and thrombocytes did not stain. Leucocytes and thrombocytes were negative for alkaline phosphatase and Sudan black B staining. Erythrocytes were negative for all cytochemical staining. The cytochemical and ultrastructural features of peripheral blood cells of G. eckloni were similar to those of other fish species. However, some important differences were identified in G. eckloni., (© 2016 Blackwell Verlag GmbH.)

Introduction: Bobtail lizards (Tiliqua rugosa) are native to Australia. The only previous study on the hematology of this species documented just 6 animals., Objectives: The aims of this study were to characterize the light microscopy, ultrastructure and cytochemistry of blood cells, and evaluate CBCs of captive Bobtail lizards., Methods: Over 2 consecutive summers, heparinized venous blood was collected from the ventral coccygeal vein of 46 clinically healthy, captive indoor- or outdoor-housed adult Bobtails. Complete blood cell counts and smear evaluations were performed, and cytochemical stains and transmission electron microscopy were used to further characterize blood cells., Results: The eosinophils of this species were uniformly vacuolated: a unique feature not previously reported in reptiles. Heterophils were the predominant leukocyte, with fewer lymphocytes, azurophilic and nonazurophilic monocytes, occasional eosinophils, and basophils. Thrombocytes were frequently clumped. Slight polychromasia (0-15% of erythrocytes) was typically present. Hemogregarine parasites were seen on some smears. The range of CBC results was often wide. The PCV ranged from 11% to 38%. Total plasma proteins by refractometry were between 3.5 and 7.8 g/dL. Hemoglobin ranged between 2.6 and 12.6 g/dL by the modified hemoglobin-hydroxylamine method. Manual RBC count was 0.35-1.27 × 10 6 /μL, and WBC count was 2.86-22.66 × 10 3 /μL. Bobtail lizards housed outdoors had lower PCVs than indoor-housed animals. Bobtails with hemogregarine infections had lower PCVs than noninfected lizards., Conclusions: Ranges for CBC data were often very wide, influenced by preanalytic and analytic factors. Hemogregarine infection is associated with a decreased PCV, suggesting that some hemogregarine species are pathogenic in this population., (© 2016 American Society for Veterinary Clinical Pathology.)

The role of goblet cell secretion, containing mucopolysaccharides, in the formation of a protective barrier of intestinal mucosa and transportation of the intestinal content has been described quite extensively. However, information on the quality composition of mucopolysaccharides and its changes in the intestinal tract of ostrich chicks, especially in the large intestinal segments, is unavailable. In the current study, ostrich embryos/chicks (n = 6/36) of both sexes were used shortly before hatching and during the first months of the post-hatch period. Tissues for histology were taken from the large intestine: the medium segments of the caecum, proximal and distal parts of colon. By using histochemical reactions, the differentiation of goblet cells as well as chemical composition of mucopolysaccharides was carried out. The cells contained acid (AB+), neutral (PAS+) and mixed (AB/PAS+) mucopolysaccharides. The number of goblet cells in the large intestine per unit area of mucosa increased towards the cloaca, and it was the highest in the distal part of the colon. The qualitative goblet cell composition in different large intestinal parts was different in all ages. In the caecum, goblet cells containing acid and mixed mucopolysaccharides dominate post-hatch, whereas in the colon, goblet cells containing acid mucopolysaccharides predominated. The most rapid changes in the qualitative goblet cell composition occur during the first week post-hatch when in all the intestinal segments the proportion of cells containing acid mucopolysaccharides continuously increased., (© 2015 Blackwell Verlag GmbH.)

The aim of the present study was to investigate the effect of gender on the histological and histochemical structures of different anatomical regions of the kidney in Japanese quail (Coturnix japonica). Tissue samples from cranial, middle and caudal divisions of each kidney were obtained from 20 male and 20 female adult, healthy Japanese quail. The sections stained with hematoxylin & eosin ( H & E: ), Masson's trichrome, Verhoeff's, Alcian blue (pH 2.5), Periodic acid-Schiff, and Gomori's method for reticulum. Unusual findings of the kidney in Japanese quail were the presence of three types of nephrons, all the connective tissue fibers in capsule and interlobular septa and AB-reactions of the proximal convoluted cells. No significant sex-based differences were found. The various histological structures of the kidney showed no significant differences among different divisions of the left and right kidneys. It is concluded that the general histological and histochemical properties of the kidney in Japanese quail were similar to those of chickens and some other species, but that there were also some differences. One of the major differences was brush border of interdigitating microvilli on luminal surface of collecting ducts in Japanese quail., (© 2016 Poultry Science Association Inc.)

Objective: To evaluate effects of laser treatment on incisional wound healing in ball pythons (Python regius)., Animals: 6 healthy adult ball pythons., Procedures: Snakes were sedated, a skin biopsy specimen was collected for histologic examination, and eight 2-cm skin incisions were made in each snake; each incision was closed with staples (day 0). Gross evaluation of all incision sites was performed daily for 30 days, and a wound score was assigned. Four incisions of each snake were treated (5 J/cm(2) and a wavelength of 980 nm on a continuous wave sequence) by use of a class 4 laser once daily for 7 consecutive days; the other 4 incisions were not treated. Two excisional skin biopsy specimens (1 control and 1 treatment) were collected from each snake on days 2, 7, 14, and 30 and evaluated microscopically. Scores were assigned for total inflammation, degree of fibrosis, and collagen maturity. Generalized linear models were used to investigate the effect of treatment on each variable., Results: Wound scores for laser-treated incisions were significantly better than scores for control incisions on day 2 but not at other time points. There were no significant differences in necrosis, fibroplasia, inflammation, granuloma formation, or bacterial contamination between control and treatment groups. Collagen maturity was significantly better for the laser-treated incisions on day 14., Conclusions and Clinical Relevance: Laser treatment resulted in a significant increase in collagen maturity at day 14 but did not otherwise significantly improve healing of skin incisions.

Heads of ten healthy adult ostrich obtained from slaughter house were the constituted materials of the study. The Harderian gland (HG) was dissected out, and all of the gross morphometrical parameters including length, width and thickness as well as weight of left and right glands were recorded. Tissue sections were stained, using haematoxylin and eosin, Masson trichrome, periodic acid-Schiff and Alcian blue (pH 2.5) techniques. In ostrich, HG was an orbital organ located ventromedially around the posterior part of the eyeball. It was an oval flatted shape, light pink colour with irregular outline and was pointed in the dorsal end. Its mean length was 35.30 ± 2.84 mm and 35.55 ± 3.58 mm in left and right sides, respectively, and mean width 15.30 ± 1.20 mm and 15.65 ± 1.18 mm in left and right sides, respectively. There was no significant difference between length, thickness, weight and width of left and right glands. Histological results showed that the glandular epithelium was multilobular and compound tubuloalveolar. The gland was surrounded by a connective tissue capsule, and the epithelium was lined by simple columnar epithelial cells of varying height. The secretion of HG was mucous and the secretion type was apocrine. Mucosubstance analysis revealed that secretory units contained acidic and neutral glycoproteins. The granules within the epithelial cells lining the intralobular and inter-lobular excretory ducts of the gland were positive for periodic acid-Schiff and Alcian blue (pH 2.5)., (© 2014 Blackwell Verlag GmbH.)

Florfenicol (FLO) is a broad-spectrum antibacterial agent for treatment of bacteriosis of piglets in veterinary practice. To study the toxicity to the hematopoietic and lymphoid organs of piglets treated with a therapeutic dose of FLO, 20 healthy weaned piglets were selected and randomly divided into two groups. Piglets in the FLO group were fed with fodder supplemented with 30mg/kg BW of FLO twice a day for 10 days. Blood samples were drawn at four time points: 1 day before FLO administration and 1, 7, and 14 days post-withdrawal. Three or four piglets were euthanized at each time point post-withdrawal and tissue samples (bone marrow, thymus and spleen) were collected for fixation and cryostorage. The levels of classical swine fever virus (CSFV) antibody against the vaccine, the concentrations of Hsp70 and IL-6 in serum and Hsp70 in tissues, and the mRNA expression levels of B-cell lymphoma 2 (bcl-2) and tumor suppressor p53 were detected, the hematology of the piglets were analyzed, and the histopathology and the status of apoptosis of the hematopoietic and lymphoid organs was examined. The results showed changes in several indicators in the FLO group 1 day post-withdrawal: the concentration of red blood cells (RBCs) was decreased, and that of platelets (PLTs) was significantly lower (p<0.05); the volumes of RBC and PLT were increased; the sum of blood lymphocytes was statistically decreased (p<0.05); the concentration of IL-6 was significantly increased (p<0.05); the concentrations of Hsp70 in serum and tissues were increased; obvious atrophy of the hematopoietic cell lines and partial replacement by fat cells were observed in bone marrow; thymus and spleen tissues showed lower concentrations and sparser arrangement of lymphocytes in the thymic medulla and white pulp of the spleen respectively; and the mRNA expression levels of bcl-2 in the three tissues were up-regulated, while that of p53 was down-regulated. With time after cessation of FLO administration, the indicators of the FLO group gradually returned to close to that of the control group and the histological lesions of the tissues gradually recovered, and the differences in the densities of lymphocytes and cell arrangements in the tissues between two groups gradually decreased. In conclusion, a therapeutic dose of FLO induces temporary toxicity in the hematopoietic and lymphoid organs of piglets to some extent, and influences hemopoiesis and immune function. These effects gradually decrease after cessation of FLO administration., (Copyright © 2014 Elsevier B.V. All rights reserved.)

Severe combined immunodeficiency (SCID) is the result of a set of inherited genetic defects which render components of the immune response nonfunctional. In Arabian horses, Jack Russell terriers, and mice, the disorder is a consequence of the absence of T and B lymphocytes, while natural killer (NK) cell and other leukocyte populations remain intact. Preliminary analysis of a naturally acquired form of inherited SCID in a line of pigs showed several defects in the architecture and composition of secondary lymphoid organs. In this study, a quantitative assessment of lymphocyte populations in affected and normal littermates showed depleted T or B lymphocyte populations in affected pigs; however, NK cells and neutrophils were present in numbers comparable to unaffected littermates. The results indicate that the immune defect in pigs shares the same features as other SCID-affected species., (Copyright © 2014 Elsevier B.V. All rights reserved.)

Case Report: A 5-year-old captive male diamond python (Morelia spilota spilota) was presented with a 1-month history of regurgitation and anorexia and discrete coelomic distention. Physical examination revealed a firm, immobile mass at approximately two-thirds of the snout-vent length from the front of the head. Ultrasound-guided fine needle aspirate biopsy of the mass in the region of the stomach showed necrosis with bacterial infiltration and possibly neoplastic changes. A gastroscopy was conducted, but showed grossly normal gastric mucosa, confirmed by biopsy. On exploratory coeliotomy, it was confirmed the mass involved most of the stomach wall and occluded the gastric lumen. The mass was completely excised and based on histopathology, a diagnosis of gastric adenocarcinoma was made. The snake was found dead 12 h postoperatively, but no specific cause of death was found on postmortem examination., Conclusion: Most cases of adenocarcinoma in snakes go undiagnosed. This case report illustrates that the architecture of gastric masses may lead to false-negative gastric biopsy results in snakes with neoplasia., (© 2014 Australian Veterinary Association.)

Case Report: An 11-year-old female terrier-cross underwent general anaesthesia and mechanical ventilation for insulinoma resection. Following tumour removal, three consecutive but unsuccessful attempts were made to place a wire-guided nasojejunal feeding tube (NJFT), using both nostrils, with one tube eventually left in situ in the right nostril. A final successful attempt using a smaller NJFT was made via the left nostril. On withdrawal of the previously, unsuccessfully placed NJFT, the dog developed an acute tension pneumothorax. Severe cardiorespiratory dysfunction ensued, including decreasing arterial haemoglobin O2 saturation, increasing end-tidal CO2, hypotension and tachycardia. Immediate thoracocentesis with air evacuation from the pleural cavity followed by continuous air drainage through bilateral thoracostomy tubes produced marked improvement of cardiorespiratory function. During exploratory thoracotomy performed 1 day later, two lung lesions were identified and over sewn, one in the right middle lobe and the other in the accessory lobe., Conclusions: This is the first case report to describe in an anaesthetised and mechanically ventilated dog the occurrence and management of a tension pneumothorax as a life-threatening complication secondary to parenchymal injury after NJFT misplacement into the tracheobronchial tree. It illustrates that presence of a cuffed endotracheal tube does not protect against passing a NJFT into the bronchial system., (© 2014 Australian Veterinary Association.)

A widespread deadly outbreak occurred in captive birds belonging to the family Turdidae in Italy. The present study was performed on 46 dead birds coming from 3 small decoy-bird breeders in central Italy. Only Turdus pilaris, Turdus iliacus, Turdus philomelos and Turdus merula were affected. No other species of bird held by these breeders died. A change of diet before the hunting season was reported from all breeders. Full necropsy of the animals and histological investigations of representative tissue samples were performed. Microscopical examination showed marked iron deposits in liver samples. Bacteriological investigations and molecular analysis to exclude bacterial and viral diseases were carried out. Contamination of food pellet samples by mycotoxins and analysis to detect heavy metal contaminants in food pellet samples were considered. An interesting result was the high iron content found in food pellets. It was higher than that considered suitable for birds, especially for species susceptible to development iron storage disease (ISD). Taken together, the results suggested an outbreak of ISD caused by the high iron content of food given to the birds before the hunting season. The high mortality recorded only in species belonging to the family Turdidae suggests a genetic predisposition in the affected birds.

The corneal-protective effects of an artificial tear containing sodium hyaluronate (SH) and castor oil (CO) were evaluated on a porcine short-term dry eye model. Fresh porcine eyes with an intact cornea were treated with an artificial tear of saline, SH solution (0.1%, 0.5% or 1%), CO solution (0.5%, 1% or 5%) or a mixture solution containing 0.5% SH and 1% CO and then desiccated for 60, 90 or 180 min. To assess corneal damage, the eyes were stained with methylene blue (MB) or lissamine green (LG). The staining score of MB, absorbance of MB extracted from the cornea and staining density of LG increased significantly with increasing desiccation time in untreated and all artificial tear-treated eyes, although there were no significant differences in staining scores and absorbance of MB between eyes treated continuously with saline and 1% SH-treated ones at 60 and 90 min of desiccation or the mixture-treated eyes at 60 min of desiccation. No significant differences in the staining density of LG were also found between continuous saline-treated eyes and ones desiccated for 60 min and treated with 1% SH and the mixture. Mild cytoplasmic vacuolations were histopathologically observed in the basal and wing cells in eyes desiccated for 60 min and treated with 1% SH and the mixture. The mixture solution containing 0.5% SH and 1% CO has protective effects against corneal desiccation similar to those of 1% SH and would be helpful as an artificial tear.

A 4-year and 2-month-old male capybara (Hydrochoerus hydrochaeris) was diagnosed with squamous cell carcinoma on the buttocks after chronic recurrent dermatosis. The capybara was euthanized, examined by computed tomography and necropsied; the tumor was examined histologically. Computed tomography showed a dense soft tissue mass with indistinct borders at the buttocks. Histological examination of the tumor revealed islands of invasive squamous epithelial tumor cells with a severe desmoplastic reaction. Based on the pathological findings, the mass was diagnosed as a squamous cell carcinoma. This is the first study to report squamous cell carcinoma in a capybara.

Case Series: The first two confirmed cases of Australian bat lyssavirus (ABLV) infection in horses are presented. Both cases occurred in the same week in May 2013 in paddock mates in south-east Queensland. Australia has been one of only a few countries considered free from rabies-like viruses in domestic animal species. ABLV infection had previously only been confirmed in bats and humans. All three confirmed human cases were fatal, the latest in February 2013. An additional human case of possible abortive infection in 1996 has also been reported. Both equine cases reported here resulted in euthanasia. The risks of infection across other mammalian species are still to be determined., Conclusions: These two equine cases highlight that ABLV should be considered as a differential diagnosis in animals with similar clinical presentations in Australia. There is a need for greater awareness regarding the zoonotic risk, use of personal protective equipment, pre- and post-exposure prophylactic measures and laboratory diagnostic options. The authors recommend ABLV testing for all Australian cases of progressive equine neurological disease., (© 2014 Australian Veterinary Association.)

Acanthocephalan and spargana parasites were identified within a body wall mass during exploratory surgery in a wild green tree snake. Acanthocephalan parasites have not previously been reported in this species. Surgical excision, the treatment of choice, could not be achieved because of the extensive infiltration of the coelomic cavity., (© 2014 Australian Veterinary Association.)

Anti-rotaviral activities of Sophora flavescens extract (SFE) and stevioside (SV) from Stevia rebaudiana Bertoni either singly or in various combinations were examined in vitro and in vivo using a porcine rotavirus G5[P7] strain. Combination of SFE and SV inhibited in vitro virus replication more efficiently than each single treatment. In the piglet model, SV had no effect on rotavirus enteritis, whereas SFE improved but did not completely cure rotaviral enteritis. Interestingly, combination therapy of SFE and SV alleviated diarrhea, and markedly improved small intestinal lesion score and fecal virus shedding. Acute toxicity tests including the piglet lethal dose 50, and body weight, organ weight and pathological changes for the combination therapy did not show any adverse effect on the piglets. These preliminary data suggest that the combination therapy of SV and SFE is a potential curative medication for rotaviral diarrhea in pigs. Determination of the efficacy of this combination therapy in other species including humans needs to be addressed in the future., (Copyright © 2014 Elsevier Ltd. All rights reserved.)

Inflammatory colorectal polyps (ICRPs) are commonly seen in miniature dachshund (MD) dogs; typically, multiple polyps form with severe neutrophil infiltration. ICRP is thought to be a novel form of inflammatory bowel disease (IBD), but its etiology has not been investigated. The innate immune system is implicated in the pathogenesis of both human and canine IBD. Therefore, the aim of the current study was to evaluate the messenger RNA (mRNA) expression profiles of pattern recognition receptors (PRRs) and cytokines in ICRPs. Polyp tissues were collected by colonoscopic biopsies from 24 MDs with ICRPs. Non-polypoid colonic mucosa was collected from all MDs with ICRPs and 21 clinically healthy beagles (as the controls). The expression levels of the mRNAs encoding toll-like receptors (TLRs) 1-10; nucleotide-binding oligomerization domain (NOD)-like receptors NOD1 and NOD2; and cytokines IL-1β, IL-6, IL-8/CXCL8, and TNF-α were evaluated by quantitative real-time RT-PCR. Three of the 10 well-known candidate reference genes were selected as housekeeper genes based on analyses from the GeNorm, NormFinder, and BestKeeper programs. Levels of TLR1, TLR2, TLR4, TLR6, TLR7, TLR8, TLR9, TLR10, NOD2, and all cytokines were significantly upregulated in the polyps relative to those in the controls. There was significant decrease in the expression levels of TLR3 and NOD1 in the polyp tissues compared to the non-polypoid colonic mucosa obtained from MDs with ICRPs. All upregulated PRR mRNAs were positively correlated with all proinflammatory cytokine mRNAs. This study demonstrated the dysregulation of PRRs and proinflammatory cytokines in ICRPs of MDs, which may play an important role in the pathogenesis of this disease., (Copyright © 2014 Elsevier B.V. All rights reserved.)

Case Report: A domestic pigeon developed a sterile granuloma following vaccination with an oil-adjuvant, inactivated La Sota strain of Newcastle disease. The aim of vaccination was to provide protection against pigeon paramyxovirus-1 (PPMV-1), a disease previously considered exotic to Australia. Granuloma formation is considered a rare complication of vaccination against PPMV-1 in pigeons. Clinical signs consistent with neuropathic pain became apparent during the extensive management of the granuloma, which included surgical removal of foreign material, control of the pain and inflammation and protection against antimicrobial contamination., Conclusion: PPMV-1 is now considered endemic in Australia and protecting pigeons with vaccination is important. Until a product is registered for use, vaccination remains off-label and the risk of adverse reaction, including sterile granuloma, must be considered., (© 2014 Australian Veterinary Association.)

Background: Anterior chamber collapse syndrome has been recognised in various species and is associated with early-life ocular disease or trauma. It is important to differentiate this acquired condition from a congenital malformation., Case Report: An adult female koala (Phascolarctos cinereus) was referred for assessment of buphthalmos and severe keratitis of the right eye. The degree of keratitis obstructed examination of intraocular structures. Enucleation of the affected eye was performed and the histopathological diagnosis was anterior chamber collapse syndrome and secondary glaucoma. This case contributes to the limited information available in the literature on anterior chamber collapse syndrome, a disease unique in having secondary glaucoma with minimal or no inflammation. The case also expands the literature available on ocular disease in koalas. More specifically, this is the only reported case of glaucoma, of any aetiology, in the koala., (© 2014 Australian Veterinary Association.)

The effects of l-Arg, vitamin C (VC), and vitamin E (VE) on xanthine- (XO) and NAD(P)H-oxidase (NOX) activities, and nitric oxide (NO) availability of hypoxic broilers were evaluated. Chickens were kept in wire cages with free access to feed and water. One-day-old chicks were assigned to 1 of 3 diets: control (CTL; ME 3,200 kcal/kg, CP 23%), high Arg (HA; CTL + Arg 0.8%), or high Arg plus VE and VC (AEC; HA + 200 IU of VE/kg of feed + 500 mg of VC/L of water), and grown under hypobaric hypoxia (HYP) from d 7 to 30. A fourth group of birds was fed the CTL diet and grown under normoxia (CTL-NOR). At d 30, chickens were euthanized, their lungs fixed in vivo, excised, and processed for cyto- and histochemistry. The enzymes XO and NOX were localized and activities assessed histochemically and in lung homogenates. The NO depletion was assessed through nitrotyrosine immunocytochemistry colloidal gold particles (NTY). The XO and NOX localized in cell membranes and within vesicles of pulmonary vessel endothelial cells. The XO activity was higher in CTL-NOR birds (586 ± 43 reflectance units) than in both AEC-HYP (456 ± 39) and HA-HYP birds (394 ± 31), whereas CTL-HYP birds had the lowest XO activity (313 ± 27). The NO depletion was not affected by dietary or hypoxia conditions in clinically healthy birds; nevertheless, hypoxic birds that developed pulmonary hypertension had higher NTY levels (less NO, 145 ± 19) than hypoxic but clinically healthy birds (56 ± 11). Thus, the concurrent supplementation of Arg, VE, and VC restored XO activity without affecting NOX activity or NO availability. The dual role of XO, which produces superoxide and uric acid, may have buffered the effects of superoxide in broiler chickens grown under hypobaric hypoxia.

Background: Canine leishmaniosis (CL) caused by Leishmania infantum is a disease of worldwide importance, not only because it causes severe and potentially fatal disease in dogs, but also because of its zoonotic relevance. The parasite is the causative agent of human visceral leishmaniosis, a severe, debilitating disease that causes an estimated 59,000 deaths annually. Australia is considered to be free of zoonotic leishmaniosis., Methods: A retrospective case series of five imported dogs diagnosed with CL between 2000 and 2011. Cases were identified by word-of-mouth and by referral., Results: The dogs were diagnosed with CL between 2000 and 2011; clinical, clinicopathological, and serological data are presented, together with a review of the disease and its biosecurity implications for Australia., Conclusions: Because of the unique immunopathology and diagnostic challenges associated with CL, the importance of obtaining a travel history is reinforced because some dogs imported prior to 2006 may develop clinical signs of this disease and present for veterinary examination. Furthermore, it is possible for leishmaniosis to become established within Australia under certain circumstances., (© 2014 Australian Veterinary Association.)

Case Report: Perennial ryegrass toxicosis (PRGT) is a common disease entity in Australia, presenting as an association of clinical signs including alterations in normal behavioural, ataxia ('staggers'), ill thrift and gastrointestinal dysfunction ('scours'). Clinical signs can range in severity from mild (gait abnormalities and failure to thrive) to severe (seizures, lateral recumbency and death). Presentation across the flock is usually highly variable. PRGT is caused by toxins produced by the endophytic fungus Neotyphodium lolii, a symbiont of perennial ryegrass that is present in pastures across the temperate regions of Australia and Tasmania. A particular feature of PRGT in Australia is the occasional occurrence of large-scale sheep losses, suggesting other factors are influencing mortality rates compared with other PRGT risk zones such as North America and New Zealand. During 2011, producers in the state of Victoria experienced a mild outbreak of PRGT that affected large numbers of animals but with limited mortalities. Clinical samples taken from affected sheep showed a high incidence of dehydration and electrolyte abnormalities., Conclusion: We speculate that changes in hydration status may be a contributory aetiological factor in those years in which high numbers of deaths are associated with PRGT outbreaks in Australia., (© 2014 Australian Veterinary Association.)

Case Report: Metaphyseal osteopathy (MO) was diagnosed in three Australian Kelpie puppies that were presented for veterinary assessment of lameness. The three puppies were siblings. Each was from a different litter by the same breeding pair. The puppy in case one was seen by the authors, and the puppies in cases two and three were patients at other veterinary hospitals. However, the medical records and radiographs were examined and reviewed for this report. Radiographic investigation of the lameness revealed pathognomonic appearance of MO affecting the metaphyseal region of the long bones in all three puppies. The diagnosis was confirmed on histopathology in one patient., Conclusion: MO is considered a disease of large and giant-breed dogs, being rarely reported in non-large-breed dogs, and has not been reported in the Australian Kelpie, which is considered a medium-breed dog. This case series suggests a previously unreported breed predisposition to MO in the Australian Kelpie., (© 2014 Australian Veterinary Association.)

Bovine Respiratory Disease Complex (BRDc), a multi-factorial disease, negatively impacts the cattle industry. Nitric oxide (NO), a naturally occurring molecule, may have utility controlling incidence of BRDc. Safety, bioavailability, toxicology and tolerance/stress of administering NO to cattle is evaluated herein. Thirteen, crossbred, multiple-sourced, commingled commercial weaned beef calves were treated multiple times intranasally over a 4 week period with either a nitric oxide releasing solution (treatment) or saline (control). Exhaled NO, methemoglobin percent (MetHg) and serum nitrites demonstrated biological availability as a result of treatment. Cortisol levels, tissue nitrites, behavior and gross and macroscopic pathology of organs were all normal. Moreover, preliminary in vitro studies using Mannheimia haemolytica, Infectious Bovine Rhinotracheitis, Bovine Parainfluenza-3 and Bovine Respiratory Syncytial Virus, suggest a potential explanation for the previously demonstrated efficacy for BRDc. These data confirm the bioavailability, safety and lack of residual of NO treatment to cattle, along with the bactericidal and virucidal effects., (Copyright © 2013 Elsevier Ltd. All rights reserved.)

Humic acids are used to prophylactically treat intestinal diseases in a wide number of species, yet the mechanism of action remains unknown. The general assumption has been that humic acids act locally; however studies using young piglets show orally supplemented humic acids can penetrate the intestinal wall, and thus potentially act systemically. The objective of this study was to determine if humic acids could also cross the intestinal barrier in adult pigs and be detected in other organs. Adult minipigs (>18 months old) orally received either 1g humic acids/kg body weight (verum, n=3) or placebo (control, n=3), for 2 weeks. At the end of the feeding period tissue samples were harvested from the intestine, various glands and organs. Unstained tissue samples were examined by light microscopy for the presence of humic acid particles. No humic acid particles were detected in any of the unstained tissues from verum or control pigs., (Copyright © 2014 Elsevier Ltd. All rights reserved.)

Adjuvants are substances added to vaccines to enhance the immune response of a given antigen. Most of the adjuvants are toxic at certain doses, and toxicity varies in different species. Moreover, there are no standard dosage limits set for adjuvant use in fish vaccines. We evaluated the acute toxicity, serum enzymes (AST/ALT) indicating hepatic injury and histopathological changes due to intra-peritoneal administration of different concentrations of a panel of adjuvants including quillaja saponin, aluminum hydroxide, squalene emulsion and Freund's incomplete adjuvant (FIA) with a dose ranging study of saponin (500, 160, 50, 16 and 5μgfish(-1)), aluminum hydroxide (5000, 1600, 500, 160 and 50μgfish(-1)), squalene emulsion (20, 10 and 5%), and FIA to determine the acceptable dosage for vaccination in olive flounder (Paralichthys olivaceus) fingerlings measuring 4.66±0.41g, 8.47±0.42cm. Saponin was highly toxic with a LD50 of approximately 105μgfish(-1) (22.4mgkg(-1)) causing severe histological damage and AST level was high at dose above 16μgfish(-1) and ALT, specific for liver damage was high only at 160μgfish(-1) (11U/L) and was safe at 5μgfish(-1). Aluminum hydroxide was toxic at 5000μgfish(-1) and was acceptable at dose below 1600μgfish(-1) with moderate histology and AST/ALT levels similar with control. Squalene emulsion showed increased inflammation at 20% and 10% emulsions and the inflammatory response was mild at a concentration of 5% oil emulsion and AST/ALT levels being similar to control in 10% and 5% emulsions and elevated in 20% on both sampling days. FIA was not lethal, but induced severe inflammation at injection site and around blood vessels. In comparison to FIA, saponin found to be safe at dose of 5μgfish(-1), aluminum hydroxide below 1600μgfish(-1), and squalene at 5% emulsion and could be accepted for vaccination studies. These results provide an insight for the selection of safer dose of adjuvants for intra-peritoneal vaccination of olive flounder., (Copyright © 2013 Elsevier B.V. All rights reserved.)

Lactobacillus brevis ATCC 8287, a surface (S-layer) strain, possesses a variety of functional properties that make it both a potential probiotic and a good vaccine vector candidate. With this in mind, our aim was to study the survival of L. brevis in the porcine gut and investigate the effect of this strain on the growth and immune function of recently weaned piglets during a feeding trial. For this, 20 piglets were divided evenly into a treatment and a control group. Piglets in the treatment group were fed L. brevis cells (1×10(10)) daily for three weeks, whereas those in the control group were provided an equivalent amount of probiotic-free placebo. For assessing the impact of L. brevis supplementation during the feeding trial, health status and weight gain of the piglets were monitored, pre- and post-trial samples of serum and feces were obtained, and specimens of the small and large intestinal mucosa and digesta were collected at slaughter. The results we obtained indicated that L. brevis-supplemented feeding induced a non-significant increase in piglet body weight and caused no change in the morphology of the intestinal mucosa. L. brevis cells were found to localize mainly in the large intestine, but they could not be isolated from feces. To a lesser extent, L. brevis was detected in the small intestine, although there was no specific attachment to the Peyer's patches. Changes in total serum IgG and IgA concentrations were not caused by supplemented L. brevis and no measurable rise in L. brevis-specific IgG was observed. However, analysis of cytokine gene expression in intestinal mucosa revealed downregulation of TGF-β1 in the ileum and upregulation of IL-6 in the cecum in the L. brevis-supplemented group. Based on the results from this study, we conclude that whereas L. brevis appears to have some intestinal immunomodulatory effects, the ability of this strain to survive and colonize within the porcine gut appears to be limited., (Copyright © 2013 Elsevier B.V. All rights reserved.)