Lead uptake into calcified and keratinized compartments of horns from a convenience sample of lead-dosed goats.

Hair and/or nail analyses are sometimes used in biomonitoring studies due to the convenience of sample collection, storage, and transport, as well as the potential to assess past exposures to toxic metals, such as lead (Pb). However, the validity of Pb measurements in these keratinized matrices as biomarkers of absorbed dose remains unclear. The aim of this study was to examine the uptake of Pb into horns harvested postmortem from 11 goats that received a cumulative oral dose of up to 151 g Pb acetate over a period of 1-11 years as part of a long-term blood Pb proficiency testing program. Uptake of Pb into keratinized horn was compared to the corresponding underlying bony horn core, which, as part of the bone compartment, provided a measure of absorbed Pb dose. Two complementary analytical techniques were used to assess Pb: X-Ray Fluorescence (XRF) and Inductively Coupled Plasma Mass Spectrometry (ICP-MS). Detectable amounts of Pb were found in all keratinized horn samples (0.45-6.6 µg/g) and in all but one bony core sample (1.4-68 µg/g). In both bony core and keratinized horn samples, Pb accumulation increased with dose over a low-to-moderate cumulative-dose interval, consistent with previous observations, but plateaued at higher doses. Significant associations were observed between Pb in keratinized horn and bony core samples particularly with XRF measurements, which represent the surface bone compartment. These findings provide evidence that Pb is excreted in keratinized tissues but reflects only a small fraction of the absorbed Pb dose, likely transferred from underlying bone tissue.

A Review of the Parasite Fauna of the Black-Bellied Pangolin, Phataginus tetradactyla LIN. (Manidae), From Central Africa with the Description of Intraproboscis sanghae n. gen., n. sp. (Acanthocephala: Gigantorhynchidae).

A new archiacanthocephalan in the family Gigantorhynchidae, Intraproboscis sanghae n. gen., n. sp. is described from females collected from the African black-bellied pangolin Phataginus tetradactyla Linn. (Manidae) in the Central African Republic. A dichotomous key to the genera of Gigantorhynchidae is provided. The specimens presented are distinct from those of the genus Gigantorhynchus Hamann, 1892 that have only 1 or 2 circles of hooks (crowns) at the apical end of the proboscis and are found in South American mammals, except for Gigantorhynchus pesteri Tadros, 1966 from baboons in Rhodesia (Zimbabwe), Africa (Amin, 2013). They superficially resemble those of the other gigantorhynchid genus Mediorhynchus Van Cleave, 1916, especially in the organization of the truncate-cone proboscis and the position of the receptacle. Species of Mediorhynchus are bird parasites. The new genus, Intraproboscis, now the third genus in Gigantorhynchidae; however, is distinguished from Mediorhynchus by having a simple proboscis receptacle that is completely suspended within the proboscis, the passage of the retractor muscles through the receptacle into the body cavity posteriorly, absence of neck, and presence of a parareceptacle structure (first finding in the Archiacanthocephala) and a uterine vesicle; among other features, including the differential dorsoventral thickness of the body wall. The receptacle in Mediorhynchus is complex, with many accessory muscles and retractor muscles passing into the body cavity dorsally and ventrally. Our specimens reached 180 mm in length and the proboscis had 34-36 rows of 6-7 ventrally lamellated, rooted hooks each anteriorly, and 15-17 spinelike hooks each posteriorly. Micropores extended into the anterior and posterior proboscis and energy dispersive x-ray analysis (EDXA) of anterior hooks showed high levels of calcium and phosphorus but negligible traces of sulfur. Spinelike hooks in the posterior proboscis had lower levels of Ca and P and slightly higher levels of S. Molecular and phylogenetic analyses based on the 18S rDNA gene placed I. sanghae in a clade with the archiacanthocephalans Mediorhynchus, Moniliformis, Macracanthorhynchus, Oncicola, and Oligacanthorhynchus.

The use of portable X-ray fluorescence spectrometry to measure apparent total tract digestibility in beef cattle and sheep.

The use of portable X-ray fluorescence (PXRF) spectrometry to detect external markers on processed or unprocessed cattle and sheep fecal specimens to estimate apparent total tract digestibility (ATTD) was evaluated. Exp. 1: ruminally cannulated Angus-crossbred steers (n = 7; BW = 520 ± 30 kg) were individually fed ad libitum for 21 d in a completely randomized design (CRD). Markers (Cr2O3 and TiO2) were placed inside the rumen twice daily (7.5 g of each marker). Fecal samples were collected twice daily from day 14 to 21. Exp. 2: crossbred wethers (n = 8; BW = 68 ± 3 kg) were individually fed ad libitum for 21 d in a CRD. During this period, 2 g of Cr2O3 and TiO2 were top-dressed onto the feed twice daily. Sheep were housed in metabolism crates for 5 d for total fecal collection. Concentration of markers was determined on diets, refusals, and fecal specimens (fresh, dry-only, and dried/ground) using atomic absorption to detect Cr and spectrophotometry for Ti. Concentration of both markers was also determined via the PXRF spectrometer. Delta between ATTD estimated by wet chemistry and PXRF was not different from zero (P ≥ 0.14) when using cattle fresh fecal specimens for both markers, whereas ATTD estimated by PXRF with dry-only and dried/ground fecal specimens were 3.6 and 1.1 percent units lower (P ≤ 0.04), respectively, than ATTD estimated by wet chemistry for Cr and Ti, respectively. Regardless of the fecal sample preparation method on cattle specimens, Ti concentration was similar (P = 0.39) among methodologies, while Cr was underestimated (P < 0.01) by 13% when PXRF was used in dry-only or dried/ground samples. The ATTD of sheep was underestimated (P < 0.01) by 2.4 percent units compared with control when Cr was measured by PXRF in dry-only samples. The Cr concentration in dry-only fecal specimens of sheep tended (P = 0.09) to be lower compared with wet chemistry analysis. Fresh and dry/ground sheep fecal samples assessed for Cr, and dry-only assessed for Ti were not (P ≥ 0.49) affected by detection method. The Cr fecal recovery tended (P = 0.10) to be the lowest for dry-only, the greatest for wet chemistry, intermediate for fresh and dry/ground sheep-fecal specimens; while not affected (P = 0.40) for Ti. The PXRF is an accurate technology to detect Cr and Ti in fresh cattle fecal samples to estimate ATTD. For fresh and dry/ground, the technology was effective for determining the concentration of Cr, or dry-only fecal specimens when detecting Ti in sheep specimens.

Morphological and Molecular Description of Rhadinorhynchus hiansi Soota and Bhattacharya, 1981 (Acanthocephala: Rhadinorhynchidae) from Marine Fish off the Pacific Coast of Vietnam.

Rhadinorhynchus hiansi Soota and Bhattacharya, 1981 , has remained unknown since its original incomplete description from 2 male specimens collected from the flat needlefish Ablennes hians Valenciennes (Belonidae) off Trivandrum, Kerala, India. Recent collections of fishes along the Pacific coast of Vietnam in 2016 and 2017 produced many specimens of the same species from the striped bonito Sarda orientalis Temminck and Schlegel (Scombridae) off the southern Pacific coast of Vietnam at Nha Trang. We describe females for the first time, assign a female allotype status, and provide an expanded description of males from a larger collection completing missing information on hooks and hook roots, receptacle, lemnisci, cement glands, Saefftigen's pouch, and trunk spines. Specimens of R. hiansi characteristically have no dorsal spines in the posterior field of trunk spines and a long proboscis with 36-48 dorso-ventrally differentiated proboscis hooks per row becoming progressively smaller posteriorly then increasing in size near the posterior end to a maximum at the posterior-most ring. Trunk, testes, and lemnisci in our specimens were considerably larger than those reported in the original description, but the proboscis was relatively smaller. The females had long reproductive system and corrugated elliptic eggs without polar prolongation of fertilization membrane. Energy Dispersive X-ray Analysis (EDXA) demonstrates high levels of calcium and phosphorus in large gallium cut hooks and high levels of sulfur in tip cuts of large and small hooks and in spines. This EDXA pattern is a characteristic fingerprint of R. hiansi. The molecular profile of R. hiansi is described from 18S rDNA and COI genes, and phylogenetic relationships with most closely related species are discussed.

Chemical and structural composition of black pigmented supragingival biofilm of bovines with periodontitis / Composição química e estrutural do biofilme supragengival pigmentado de bovinos com periodontite

Bovine periodontitis is a multifactorial disease primarily associated with a potentially pathogenic microbiota housed in the oral biofilm of animals. Biofilms are organized structures, in which the constituents coexist in symbiosis, already described as a predisposing factor to periodontitis in other species. The objective of the present study was to characterize the structure and chemical aspects of the bovine black pigmented supragingival biofilm using scanning electron microscopy (SEM) and energy dispersive spectroscopy (EDS), respectively, and determine its relationship with bovine periodontitis. Eleven premolar teeth from different animals were evaluated; five non-pigmented samples and six samples with black pigmented biofilms were initially submitted to SEM, and three areas of these samples were selected for EDS. The structure of the pigmented biofilm was more complex and irregular because of a higher content of mineral elements. The semi-quantitative EDS data indicated an association of iron (p<0.014) and magnesium (p<0.001) with the occurrence of periodontitis, whereas carbon, phosphorus, calcium, manganese, sodium, and potassium were not associated with the disease. Carbon (p<0.039), manganese (p<0.007), and iron (p<0.015) were associated with pigmentation, whereas phosphorus, calcium, and magnesium were not associated with it. Spearman correlation test showed the relationships between calcium and phosphorus, and iron and silicon. The strong association of iron in the pigmented supragingival biofilm and with the occurrence of periodontitis suggests the presence of microorganisms that use this element in their metabolism and that are also associated with bovine periodontitis. This study suggests that the pigmented deposits in the crown of the teeth of cattle are an true biofilm with the deposition of iron, and it indicates that the presence of iron and magnesium in these formations may be involved in the metabolism of some microorganisms associated with the etiology of bovine periodontitis.(AU)

A periodontite bovina é uma infecção multifatorial associada primariamente à microbiota potencialmente patogênica presente no biofilme bucal. Biofilmes são estruturas organizadas, nas quais os constituintes convivem em simbiose, descritos em outras espécies como um fator predisponente à periodontite. O objetivo do presente estudo foi caracterizar estrutural e quimicamente o biofilme supragengival pigmentado de preto em bovinos, utilizando-se as técnicas de microscopia eletrônica de varredura (MEV) e espectroscopia de dispersão de energia (EDS), respectivamente, correlacionando os elementos identificados à ocorrência de periodontite e pigmentação. Foram avaliados 11 dentes primeiro-molares; cinco amostras sem pigmentação visível e seis amostras com biofilme pigmentado de preto, que foram submetidas inicialmente à MEV; posteriormente foram selecionadas três áreas aleatórias de cada dente para realização da EDS. A estrutura do biofilme pigmentado revelou formações irregulares e mais complexas, provavelmente devido ao maior acúmulo de elementos minerais. Os resultados semi-quantitativos da EDS apontaram associações entre a presença de ferro (p<0,014) e magnésio (p<0,001) com a ocorrência de periodontite. Carbono, fósforo, cálcio, manganês, sódio e potássio não apresentaram associação com a periodontite. Em relação à pigmentação, carbono (p<0,039), manganês (p<0,007) e ferro (p<0,015) foram os elementos estatisticamente significantes, enquanto fósforo, cálcio e magnésio não apresentaram associação com a pigmentação. O teste de correlação de Spearman demonstrou associações entre os elementos cálcio e fósforo, e ferro e silício. A forte associação do ferro presente no biofilme supragengival com a ocorrência de periodontite, sugere a presença de micro-organismos que utilizam este elemento em seu metabolismo e que possivelmente tenham envolvimento com o desenvolvimento da periodontite bovina. Os resultados inéditos do presente trabalho sugerem que os depósitos pigmentados que se formam na coroa dos dentes de bovinos são um biofilme verdadeiro com deposição de ferro, e indicam que a presença de ferro e magnésio nestas formações pode estar envolvida no metabolismo de alguns dos principais micro-organismos associados à etiologia da periodontite bovina.(AU)

Bovine eggs release zinc in response to parthenogenetic and sperm-induced egg activation.

Upon fertilization or parthenogenesis, zinc is released into the extracellular space through a series of exocytic events termed zinc sparks, which are tightly coordinated with intracellular calcium transients. The zinc spark reduces the total amount of intracellular zinc, and this reduction is necessary and sufficient to induce egg activation even in the absence of calcium transients. In addition, this zinc release contributes to the block to polyspermy through modification of the zona pellucida. The zinc spark has been documented in all organisms examined to date including the mouse, two species of nonhuman primates, and human. Here we determined whether zinc sparks occur in the bovine, an important model of gamete development in mono-ovulatory mammalian species. We obtained metaphase II-arrested (MII) bovine eggs following in vitro maturation. Total zinc, assessed in single cells using X-Ray Fluorescence Microscopy, was significantly more abundant in the bovine egg compared to iron and copper. Studies with intracellular fluorescent probes revealed that labile zinc pools are localized to discrete cytoplasmic punctae enriched at the cortex. To determine whether zinc undergoes dynamic fluxes during egg activation, we parthenogenetically activated bovine eggs using two approaches: ionomycin or bovine phospholipase C zeta (bPlcζ). Both these methods induced zinc sparks coordinately with intracellular calcium transients. The zinc spark was also observed in bovine eggs following intracytoplasmic sperm injection. These results establish that zinc is the most abundant transition metal in the bovine egg, and zinc flux during egg activation - induced by chemical activation or sperm - is a highly conserved event across mammalian species.

Technical note: Rapid mineral determination in forages by X-ray fluorescence.

A large portion of the cow's ration is composed of forages that can vary greatly in mineral concentrations, which may affect animal performance and health. Current methods for mineral analysis require sample destruction either through wet or dry ashing and complex analytical techniques for individual minerals. Energy dispersive X-ray fluorescence (EDXRF) is a nondestructive, multi-mineral, spectroscopy technique, which makes mineral quantification simple, direct, and affordable. The study objective was to evaluate the prediction performance of EDXRF of Na, Mg, P, S, Cl, Ca, K, Mn, Fe, Cu, and Zn concentrations in forages. Twelve certified plant samples with a wide range in mineral concentrations were used to develop calibrations, and 35 forages (18 alfalfa hays, 10 grass hays, 7 corn silages) with measured mineral concentrations, which were collected over several years from 3 proficiency programs, were used as an independent validation set. All the samples were previously dried and finely ground and were prepared by compressing them into a round dense pellet, 40 mm in diameter, using a 40-ton pneumatic laboratory press. Samples were scanned using an EDXRF instrument enhanced for lighter minerals like Na and Mg. Samples were scanned at 20 keV and at 40 KeV associated with an Al filter, for a total analysis time of approximately 6 min. Calibrations were developed with Bruker SpectraEDX (Bruker, Hamburg, Germany) software and optimized to minimize the standard error of calibration. All of the minerals had acceptable calibration performance with coefficient of determination ranging from 0.93 (P) and 0.99 (Cl, Ca, and Mn) and coefficients of variation within 5 to 14%, which are similar to the coefficients of variation of the reference analysis. The coefficients of variation for Na was an exception, with a coefficients of variation of 29%. The validation set obtained similar statistical results as that observed in calibration. The root mean square error of prediction corrected for bias was similar to the standard error of calibration, indicating that it is possible to build a robust calibrations that performed well across different type of forages by using 12 reference samples with a sufficient range in mineral concentrations that were determined accurately. A bias correction was necessary to improve prediction accuracy only for K (-0.23% dry matter) and Ca (-0.16% dry matter). Energy dispersive X-ray fluorescence demonstrated the ability to be an accurate, direct, and simple technique for forage mineral analysis.

Morphology and elemental analysis of free range and stabled Polish Konik horses hair using Energy-dispersive X-ray spectroscopy (EDS).

Hair is a more biologically stable material than other tissues and contains a relatively constant level of minerals, drugs or toxins. The content of essential elements in hair depends on dietary and non-dietary factors. The aim of this study was to assess and compare the effect of diet on the elemental composition and morphological properties of the hair of healthy Polish Konik horses. Mane hair was collected from 19 horses living in The National Park in Popielno in Poland. Six of these horses were free-ranged all year with permanent access to pasture, while 13 horses were kept in the stable and fed oats, hay and straw. The samples were analyzed using a Scanning Electron Microscope combined with Energy-dispersive X-ray spectroscopy (SEM-EDS). Each hair was analyzed for its microscopic appearance and elemental Mg, Si, S, Ca, Mn, Co, Zn and Se content. No significant changes in the morphological properties and elemental composition were observed. There was inter- individual variability in element content between horses in both groups, but this did not affect their health. The comparison of the present results with other studies shows that the level of elements in the hair of Polish Konik horses is significantly lower than in healthy horses of other breeds. The traditional feeding plan containing pasture, oats and hay does not contain a sufficient amount of main mineral elements. Further investigation is needed to explain the effect of diet and maintenance on the mineral balance of horses.

Cuticle and pore plug properties in the table egg.

Food safety of table eggs is vital since many pathogens can contaminate the unfertilized egg, leading to increased risk of foodborne illness for consumers. The eggshell cuticle is the first line of defense to restrict the entry of egg-associated pathogens, such as Salmonella Enteritidis. The thickness and completeness of coverage of the cuticle layer are heritable traits that are strongly associated with egg resistance to bacterial penetration. The present study characterizes the chemical composition of the eggshell cuticle and structure of pore plugs from table eggs. Eggs collected from both brown and white egg laying Lohmann flocks (early, mid, and late lay) were either unwashed or washed. Pore plugs were characterized using scanning electron microscopy (SEM), and elemental composition was determined using energy-dispersive x-ray spectroscopy (EDS). SEM observations confirmed that the plug formed by the cuticle layer within the eggshell pore remains firmly lodged throughout the commercial washing process. The eggshell thickness and cuticle pore length visualized in brown eggs was significantly higher than in white eggs in hens of all ages. EDS analysis revealed that the pore inner surface was enriched in phosphorus and chemically different from the surrounding bulk eggshell mineral. Detailed assessment of the cuticle chemical composition was performed by Fourier transform infrared spectroscopy (FTIR). Washing of eggs removed cuticle from the eggshell surface. There was a trend of lower cuticle coverage with increasing hen age for white eggs. A significant reduction in the amount of proteins and phosphates and polysaccharides was observed in the cuticle of brown unwashed eggs with hen age. In white unwashed eggs, amides and lipids decreased with hen age; by contrast, the amount of sulfate was highest at mid-lay. The results from our research will assist selective breeding programs that target cuticle integrity and pore plug stability to enhance egg resistance to pathogen penetration and improve food safety.

Rhadinorhynchus oligospinosus n. sp. (Acanthocephala, Rhadinorhynchidae) from mackerels in the Pacific Ocean off Peru and related rhadinorhynchids in the Pacific, with notes on metal analysis.

Specimens of a new species of Rhadinorhynchus Lühe, 1911 are described from the chub mackerel Scomber japonicus (Scombridae) and the Chilean Jack mackerel Trachurus murphyi (Carangidae) (possibly a subspecies of Trachurus symmetricus) from the Pacific Ocean off the Peruvian coast at the Port of Chicama, La Libertad. Specimens of Rhadinorhynchus oligospinosus n. sp. are somewhat small having 11-14 rows of alternating proboscis hooks with 20-22 hooks each with posteriormost hooks in a continuous ring. Ventral hooks are robust with prominent roots but dorsal hooks are slender and shorter with discoid roots. Trunk spines are in two zones separated by a non-spiny region. Anterior trunk spines are in 2-3 complete circles but posterior spines are only ventral and lateral, and do not extend posterior to the level of the posterior end of the proboscis receptacle in both sexes. The new species is closest to Rhadinorhynchus seriolae (Yamaguti, 1963) Golvan, 1969 found in Japanese and Australian waters, but not as close to 19 other species found in the same Pacific waters off Australia, Japan, and Vietnam. In R. seriolae, posterior trunk spines extend well past the receptacle in females, among other diagnostic differences. Proboscis hooks of the new species were analyzed for chemical elements using X-ray in conjunction with EDAX (energy-dispersive analysis for X-ray) software; sulfur had a higher concentration at the edge than the middle of cut hooks.

Redescription of Pallisentis (Brevitritospinus) Indica (Acanthocephala: Quadrigyridae) from Channa punctatus Bloch & Schneider (Channidae) in Aligarh, India with New Understandings of Old Structures.

Pallisentis indica Mital and Lal, 1976 was originally described from Channa gachua Hamilton (Channidae) in Kali Nadi River, Aligarh, India. The parasite was later placed in the subgenus Brevitritospinus Amin, Heckmann, Ha, Luc, and Doanh, 2000 . Our collection from the spotted snakehead Channa punctatus Bloch & Schneider in another locality of the same stream in Aligarh produced many specimens with variable traits, revealing new structures that have never before been described in the Acanthocephala, especially relating to the ducted trunk spines. The proboscis has 4 circles of 10 hooks each, with hooks in the anterior 2 circles being considerably larger than those in the posterior 2 circles. Y-shaped trunk spines are ducted in 2 regions separated by a spineless zone. The anterior collar spines are in complete rings of 9-17 circles of crowded spines and the larger posterior trunk spines are in 1 (posterior) to 41 (anterior) circles extending to level of cement glands in males posteriorly. Considerable variations from the original description and new structures are reported for the first time.

Engineered nanoparticles of titanium dioxide (TIO2): Uptake and biological effects in a sea bass cell line.

With the rapid development of nanotechnology there has been a corresponding increase in the application of titanium dioxide nanoparticles (TiO2-NPs) in various consumer and industrial products, consequently their potential health hazards and environmental effects are considered an aspect of great concern. In the present study, in order to assess the impact of TiO2-NPs in the marine environment, the biological effects of TiO2-NPs on a sea bass cell line (DLEC) were investigated. Cells were exposed for 24 h to different concentrations of TiO2-NPs (1, 8, 40, 200 and 1000 µg/ml) or co-exposed with CdCl2 (Cd). The effects of UV light irradiation were also investigated in cells treated with TiO2-NPs and/or Cd. The internalization of TiO2-NPs and the morphological cell modifications induced by the treatments were examined by transmission and scanning electron microscopy, this latter coupled with energy dispersive X-ray spectroscopy (EDS) for particle element detection. In addition, the effects of controlled exposures were studied evaluating the cytotoxicity, the DNA damage and the expression of inflammatory genes. Our study indicates that TiO2-NPs were localized on the cell surface mainly as agglomerates revealed by EDS analysis and that they were uptaken by the cells inducing morphological changes. Photoactivation of TiO2-NPs and/or co-exposure with Cd affects ATP levels and it contributes to induce acute cellular toxicity in DLEC cells dependent on Ti concentration. The inflammatory potential and the DNA damage, this latter displayed through a caspase-3 independent apoptotic process, were also demonstrated. Overall our data suggest that the interaction of TiO2-NPs with marine water contaminants, such as cadmium, and the UV irradiation, may be an additional threat to marine organisms.

Understanding irregular shell formation of Nautilus in aquaria: chemical composition and structural analysis.

Irregular shell formation and black lines on the outside of live chambered nautilus shells have been observed in all adult specimens at aquariums and zoos soon after the organisms enter aquaria. Black lines have also been observed in wild animals at sites of broken shell, but continued growth from that point returns to a normal, smooth structure. In contrast, rough irregular deposition of shell continues throughout residence in aquaria. The composition and reasons for deposition of the black material and mitigation of this irregular shell formation is the subject of the current study. A variety of analytical techniques were used, including stable isotope mass spectrometry (SI-MS), inductively coupled plasma mass spectrometry (ICP-MS), micro x-ray fluorescence (µXRF), X-ray diffraction (XRD), and scanning electron microscopy (SEM) based X-ray microanalysis. Results indicate that the black material contains excess amounts of copper, zinc, and bromine which are unrelated to the Nautilus diet. The combination of these elements and proteins plays an important role in shell formation, growth, and strengthening. Further study will be needed to compare the proteomics of the shell under aquaria versus natural wild environments. The question remains as to whether the occurrence of the black lines indicates normal healing followed by growth irregularities that are caused by stress from chemical or environmental conditions. In this paper we begin to address this question by examining elemental and isotopic differences of Nautilus diet and salt water. The atomic composition and light stable isotopic ratios of the Nautilus shell formed in aquaria verses wild conditions are presented.

Change in the chicken eggshell cuticle with hen age and egg freshness.

For a fuller understanding of the functionality of the eggshell cuticle, we conducted a detailed study using a wide array of analytical techniques (scanning and transmission microscopy), energy dispersive x-rays, and attenuated total reflection-Fourier transform infrared spectroscopy to analyze the structure, morphology, and chemical composition of this organic coating. This study shows that the cuticle has a compositional gradation with an outer part richer in proteins and an inner part richer in sulfated polysaccharides and phosphates. It also shown that the cuticle composition, thickness, and degree of coverage are highly dependent on hen age and egg freshness. During the course of the first laying year, the thickness and degree of glycosylation of the cuticle decreases with hen age, and at the end of the laying cycle, the cuticle is significantly depleted in lipids. There are also well-defined compositional changes in the cuticle of freshly laid eggs as time passes and there is a notable increase in the permeability of the eggshell after 24 h due to cuticle drying. We discuss how these changes in the cuticle can affect the food safety of eggs in relation to the risk of trans-shell contamination by bacteria (i.e., Salmonellosis).

Pathological features of oxalate nephrosis in a population of koalas (Phascolarctos cinereus) in South Australia.

The wild and captive koala population of the Mt Lofty Ranges in South Australia has a high level of renal dysfunction in which crystals consistent with calcium oxalate have been observed in the kidneys. This study aimed to describe the pathological features of the renal disease in this population, confirm the composition of renal crystals as calcium oxalate, and determine whether any age or sex predispositions exist for this disease. A total of 51 koalas (28 wild rescues, 23 captive) were examined at necropsy, of which 28 (55%) were found to have gross and/or histological evidence of oxalate nephrosis. Histopathological features included intratubular and interstitial inflammation, tubule dilation, glomerular atrophy, tubule loss, and cortical fibrosis. Calcium oxalate crystals were demonstrated using a combination of polarization microscopy, alizarin red S staining, infrared spectroscopy, and energy-dispersive X-ray analysis with scanning electron microscopy. Uric acid and phosphate deposits were also shown to be present but were associated with minimal histopathological changes. No significant differences were found between the numbers of affected captive and wild rescued koalas; also, there were no sex or age predispositions identified, but it was found that oxalate nephrosis may affect koalas <2 years of age. The findings of this study suggest that oxalate nephrosis is a leading disease in this koala population. Possible causes of this disease are currently under investigation.

Optimized radiographic spectra for small animal digital subtraction angiography.

The increasing use of small animals in basic research has spurred interest in new imaging methodologies. Digital subtraction angiography (DSA) offers a particularly appealing approach to functional imaging in the small animal. This study examines the optimal x-ray, molybdenum (Mo) or tungsten (W) target sources, and technique to produce the highest quality small animal functional subtraction angiograms in terms of contrast and signal-difference-to-noise ratio squared (SdNR2). Two limiting conditions were considered-normalization with respect to dose and normalization against tube loading. Image contrast and SdNR2 were simulated using an established x-ray model. DSA images of live rats were taken at two representative tube potentials for the W and Mo sources. Results show that for small animal DSA, the Mo source provides better contrast. However, with digital detectors, SdNR2 is the more relevant figure of merit. The W source operated at kVps >60 achieved a higher SdNR2. The highest SdNR2 was obtained at voltages above 90 kVp. However, operation at the higher potential results in significantly greater dose and tube load and reduced contrast quantization. A reasonable tradeoff can be achieved at tube potentials at the beginning of the performance plateau, around 70 kVp, where the relative gain in SdNR2 is the greatest.

Iohexol plasma clearance in healthy dogs and cats.

Iohexol plasma clearance as a measure of glomerular filtration was determined in 31 dogs and 19 cats after an intravenous (i.v.) bolus injection. All animals were healthy and privately owned. Serial blood samples were taken before and up to 4 h after tracer injection. Iohexol plasma concentration was determined using X-ray fluorescence. A plasma tracer elimination curve was generated and clearance was calculated by dividing the injected dose by the area under the curve estimated using a two-compartment pharmacological model. Clearance was normalized to body weight (BW), body surface area (BSA), and extracellular fluid volume (ECFV). Mean, SD, and coefficient of variation of plasma clearance, before and after normalization, were calculated. Linear regression analyses were performed between body size and normalized plasma clearances. No significant linear relation was found between BSA and clearance normalized to BSA in dogs, and between BSA, BW, ECFV and clearance normalized to BSA, BW, and ECFV in cats. The optimal method for normalization of iohexol plasma clearance in dogs was by using BSA. In cats, all three methods tested were considered satisfactory. Normalization to BSA appears to be superior to normalization to BW and ECFV in dogs, and can be recommended for clinical use.

Detection of silver protein complex injections in the bovine udder using X-ray fluorescence: a preliminary investigation.

To determine the feasibility of using x-ray fluorescence (XRF) to detect the presence of silver in the mammary gland of dairy cows injected with mild silver protein suspension. The XRF spectroscopy was conducted on cadaver udders with and without mild silver protein injected. Spectral analysis was performed in order to determine the amplitude of the silver K-alpha peak that was detected. By comparing the amplitude of the K-alpha peak to the background, a minimum time of collection was determined, as a measure of the time required to observe a silver signal that is significantly non-zero. The minimum detection time required for evidence of injected silver suspension was calculated to be 2.8 +/- 0.2 s. Even with an additional requirement that the net signal exceed 50 counts, the clear indication of the presence of silver will be observed within 4 min of interrogation. X-ray fluorescence spectroscopy was shown to be a viable method for the detection of injected silver protein in cadaver mammary glands of dairy cows. While these findings are promising, further studies must be conducted to investigate the time dependence of the silver signal when diffusion, absorption, and redistribution are involved, under conditions that better mimic those encountered at an exhibition. This technique, used in conjunction with screening ultrasound examinations, has the potential to confirm sites of injection of silver compounds.

Effects of sevoflurane, isoflurane and halotane anaesthesia on fluorescein angiographic phases of dogs: a comparative study.

A fluorescein angiography method was developed to compare the onset and the total duration of the fluorangiographic phases between three anaesthetic protocols in six healthy mixed-breed dogs. The animals were anaesthetized three times. Each dog received, as pre-anaesthetic protocol, atropine (10 micrograms/kg intramuscularly), and as a sedative, romifidine (80 micrograms/kg intravenously). Fifteen minutes later, induction of anaesthesia was delivered with propofol (1 mg/kg intravenously) and maintained either with sevoflurane (SEVO group), isoflurane (ISO group) or halothane (HAL group) for 30 min in all cases. Some angiographic, cardiovascular and respiratory variables were registered during the procedure. Recovery times were also registered. Angiographic variables recorded were: onset of the arterial phase (TA), onset of the arteriovenous phase (TAV), onset of the venous phase (TV), complete arterial phase duration (I1), complete arteriovenous phase duration (I2) and I1 plus I2 (I3). Mean heart rate, mean arterial pressure, systolic arterial pressure, diastolic arterial pressure, respiratory rate, tidal volume, arterial oxygen saturation and end-tidal CO2 during SEVO and ISO anaesthesia, were similar in dogs. Minute ventilation and rectal temperature were higher in dogs with SEVO than ISO. HAL produced higher arterial pressures and a lower arterial oxygen saturation than ISO and SEVO. Mean respiratory rate, rectal temperature and minute ventilation were higher in HAL. Pulse rate, end-tidal CO2 and tidal volume were similar in the dogs of the three groups. No differences in recovery times were found. The fluorescein angiographic times were within the normal range. There were no significant differences between protocols in I1, I2 or I3. HAL produced a significant increase of all temporal variables (TA, TAV and TV) when compared with ISO; TA was higher in HAL than SEVO-treated dogs. All protocols appear to be safe and effective for inducing and maintaining general anaesthesia in healthy dogs for performing fluorescein angiography.

Relationship between plasma iohexol clearance and urinary exogenous creatinine clearance in dogs.

The objective of this study was to determine if plasma iohexol clearance, computed by a 1-compartment model defined by 3 plasma samples. was an accurate measure of glomerular filtration rate (GFR) in dogs. Twenty-two adult Beagle dogs of both genders were studied. Ten dogs had intact kidneys, and 12 dogs had surgically reduced renal mass. A bolus injection of iohexol was made, and blood was obtained for plasma iohexol assay after 120, 180, and 240 minutes. Plasma was analyzed for iohexol concentration by means of 3 assay methods: chemical, high-performance liquid chromatography (HPLC), and inductively coupled plasma emission spectroscopy (ICP). Urinary clearance of exogenous creatinine was used to measure GFR for three 30-minute periods occurring between 150 and 240 minutes after iohexol injection. Plasma clearance of iohexol and renal clearance of creatinine were compared by linear regression analysis and by limits of agreement techniques. Plasma iohexol clearance and urinary exogenous creatinine clearance were significantly correlated (chemical R2 = .90; HPLC R2 = .96; and ICP R2 = .96). The 1-compartment iohexol clearance:exogenous creatinine clearance ratios were 1.04 +/- 0.17, 1.05 +/- 0.14, and 1.10 +/- 0.15 for the chemical, HPLC, and ICP methods of assay, respectively, indicating that plasma iohexol clearance slightly overestimated GFR. Assuming a +/- 2 standard deviation interval for error, corrected plasma iohexol clearance measured GFR with +/-34% accuracy for the chemical, +/-26% accuracy for the HPLC, and +/-27% accuracy for the ICP method. These results indicate that plasma iohexol clearance should have utility for detection of renal dysfunction earlier in the course of progressive renal disease than is possible with measurement of plasma creatinine or urea concentrations.