Toxicological alterations induced by subacute exposure of silver nanoparticles in Wistar rats.

Silver nanoparticles (AgNPs) have become crucial players in the field of medicine and various other industries. AgNPs have a wide array of applications, which includes production of electronic goods, cosmetics, synthesis of dyes, and printing inks, as well as targeted delivery of drugs to specialized cells inside the body. Even though humans readily come in contact with these particles, the organ-specific accumulation and resulting mechanisms of toxicity induced by inhaled AgNPs are still under investigation. The goal of this study was to determine the organ distribution of inhaled AgNPs and investigate the resulting systemic toxicity. To do this, male Wistar rats were exposed by inhalation to AgNPs for 4 hr/day (200 parts per billion/day) for five consecutive days. The nanoparticles were generated using a laser ablation technique using a soft-landing ion mobility (SLIM) instrument. Inductively coupled plasma mass spectrometric (ICP-MS) analysis showed organ-specific accumulation of the nanoparticles, with the highest concentration present in the lungs, followed by the liver and kidneys. Nanoparticle distribution was characterized in the organs using scanning electron microscopy (SEM) and matrix-assisted laser desorption/ionization mass spectrometric (MALDI-MS) imaging. Bone marrow cytotoxicity assay of the cells from the femur of rats showed micronuclei formation and signs of cellular cytotoxicity. Moreover, rats displayed increased levels of circulating lactate and glutathione disulphide (GSSG), as determined by liquid chromatography-mass spectrometry (LC-MS) analysis. Collectively, our observations suggest that inhaled subacute exposure to AgNP results in accumulation of AgNPs in the lungs, liver, and kidneys, preferentially, as well as mediates induced systemic toxicity.

Hemorrhagic diathesis in avian species following intramuscular administration of polysulfated glycosaminoglycan.

Polysulfated glycosaminoglycans (PSGAGs) have been used for decades in a variety of species for the management of osteoarthritic pain. However, reports on the use of PSGAGs in avian species are scarce. In domestic cats and dogs, PSGAG injections have caused prolongation of clotting times but are considered to be an efficacious drug with a wide margin of safety. This publication documents four cases of fatal coagulopathies in different avian species (one coraciiforme, two raptors, and one psittacine) following the intramuscular administration of PSGAG. All affected birds received varying dosages and dosing intervals of PSGAG. Three of the four birds experienced fatal hemorrhage into the pectoral muscle, while the fourth bled continuously from the injection site. Only one bird had chronic, severe pre-existing disease; the remainder were being managed for osteoarthritis. This report highlights the importance of species-specific dosing of PSGAG and warrants further investigation into the etiopathogenesis of this process.

Evolution of modern birds revealed by mitogenomics: timing the radiation and origin of major orders.

Mitochondrial (mt) genes and genomes are among the major sources of data for evolutionary studies in birds. This places mitogenomic studies in birds at the core of intense debates in avian evolutionary biology. Indeed, complete mt genomes are actively been used to unveil the phylogenetic relationships among major orders, whereas single genes (e.g., cytochrome c oxidase I [COX1]) are considered standard for species identification and defining species boundaries (DNA barcoding). In this investigation, we study the time of origin and evolutionary relationships among Neoaves orders using complete mt genomes. First, we were able to solve polytomies previously observed at the deep nodes of the Neoaves phylogeny by analyzing 80 mt genomes, including 17 new sequences reported in this investigation. As an example, we found evidence indicating that columbiforms and charadriforms are sister groups. Overall, our analyses indicate that by improving the taxonomic sampling, complete mt genomes can solve the evolutionary relationships among major bird groups. Second, we used our phylogenetic hypotheses to estimate the time of origin of major avian orders as a way to test if their diversification took place prior to the Cretaceous/Tertiary (K/T) boundary. Such timetrees were estimated using several molecular dating approaches and conservative calibration points. Whereas we found time estimates slightly younger than those reported by others, most of the major orders originated prior to the K/T boundary. Finally, we used our timetrees to estimate the rate of evolution of each mt gene. We found great variation on the mutation rates among mt genes and within different bird groups. COX1 was the gene with less variation among Neoaves orders and the one with the least amount of rate heterogeneity across lineages. Such findings support the choice of COX 1 among mt genes as target for developing DNA barcoding approaches in birds.

Ingluviotomy tube placement for lead-induced crop stasis in the California condor (Gymnogyps californianus).

Six free-flying California condors (Gymnogyps californianus) were diagnosed with acute lead toxicosis that caused crop distension and stasis. Between January 2006 and January 2007, the birds were referred to the Phoenix Zoo in Arizona for emergency treatment. In 5 birds, an ingluviotomy was performed to place a feeding tube from the crop to the proventriculus, which allowed a temporary bypass of the dysfunctional esophagus until normal function and motility were regained. A crop-support pressure bandage was placed in 4 birds to improve crop emptying into the proventriculus and to prevent crop distension. Although chelation therapy is the gold standard treatment for lead toxicosis, severe cases of lead-induced crop stasis are not acutely reversible with pharmaceuticals. In these condors, placement of a feeding tube was deemed prudent to ensure a viable enteric route of nutritional support during the standard treatment and recovery period in acute lead toxicosis with crop stasis.

Long-term treatment of chronic lymphocytic leukemia in a green-winged macaw (Ara chloroptera).

A 32-year-old green-winged macaw (Ara chloroptera) was diagnosed with chronic lymphocytic leukemia based on progressive lymphocytosis and the presence of a monomorphic population of well-differentiated lymphocytes in the bone marrow of a clinically normal bird. Chemotherapy was initiated because of rapidly increasing peripheral lymphocyte counts. In addition to oral prednisone (1 mg/kg once daily), oral chlorambucil (1 mg/kg twice weekly) was initiated but was discontinued after 6 weeks because of thrombocytopenia. The leukocyte count was stabilized for 29 weeks with the concurrent use of oral cyclophosphamide (5 mg/kg 4 d/wk) and daily prednisone, and the bird exhibited a good quality of life. The bird died shortly after the chemotherapy was inadvertently discontinued. The neoplastic cells from this macaw stained positive for CD-3 antibody and negative for Bla.36, suggesting the leukemia was of T-cell origin. This is the first report of long-term treatment of a macaw with cyclophosphamide and documents thrombocytopenia in a macaw secondary to chlorambucil treatment.

Metallic nanoparticle production and exposure/deposition system for toxicological research applications using zebrafish.

Metallic nanoparticles (NPs) have been accepted for various applications ranging from cosmetics to medicine. However, no method has been established in the scientific community that is capable of analyzing various metals, sizes, and levels of exposures without the concern of background chemical contaminations. We present here a system utilizing soft-landing ion mobility (SLIM) exposures of laser ablated metallic clusters capable of operating pressures of reduced vacuum (1 Torr) up to ambient (760 Torr) in the presence of a buffer gas. Clusters experience kinetic energies of less than 1 eV upon exiting the SLIM, allowing for the exposure of NPs to take place in a passive manner. While there is no mass-selection of cluster sizes in this work, it does show for the first time the creation and soft-landing of nanoclusters at ambient pressures. Factors such as area coverage and percentage distribution were studied, as well as the different effects that varying surfaces may cause in the agglomeration of the clusters. Furthermore, the system was successfully used to study the effects of silver nanoparticle exposure and determine the specific organs the NPs accumulate in using zebrafish (Danio rerio) as a model organism. This method provides a novel way to synthesize NPs and expose biological organisms for various toxicological analysis.

The endangered California Condor (Gymnogyps californianus) population is exposed to local haemosporidian parasites.

The endangered California Condor (Gymnogyps californianus) is the largest New World Vulture in North America. Despite recovery program success in saving the species from extinction, condors remain compromised by lead poisoning and limited genetic diversity. The latter makes this species especially vulnerable to infectious diseases. Thus, taking advantage of the program of blood lead testing in Arizona, condor blood samples from 2008 to 2018 were screened for haemosporidian parasites using a nested polymerase chain reaction (PCR) protocol that targets the parasite mitochondrial cytochrome b gene. Plasmodium homopolare (Family Plasmodiidae, Order Haemosporida, Phylum Apicomplexa), was detected in condors captured in 2014 and 2017. This is the first report of a haemosporidian species infecting California Condors, and the first evidence of P. homopolare circulating in the Condor population from Arizona. Although no evidence of pathogenicity of P. homopolare in Condors was found, this study showed that the California Condors from Arizona are exposed to haemosporidian parasites that likely are spilling over from other local bird species. Thus, active surveillance should be an essential part of conservation efforts to mitigate the impact of infectious diseases, an increasingly recognized cause of global wildlife extinctions worldwide, particularly in avian populations considered vulnerable or endangered.

O-Glycosylated 24 kDa human growth hormone has a mucin-like biantennary disialylated tetrasaccharide attached at Thr-60.

MS was used to characterize the 24 kDa human growth hormone (hGH) glycoprotein isoform and determine the locus of O-linked oligosaccharide attachment, the oligosaccharide branching topology, and the monosaccharide sequence. MALDI-TOF/MS and ESI-MS/MS analyses of glycosylated 24 kDa hGH tryptic peptides showed that this hGH isoform is a product of the hGH normal gene. Analysis of the glycoprotein hydrolysate by high-performance anion-exchange chromatography with pulsed amperometric detection and HPLC with fluorescent detection for N-acetyl neuraminic acid (NeuAc) yielded the oligosaccharide composition (NeuAc(2), N-acetyl galactosamine(1), Gal(1)). After beta-elimination to release the oligosaccharide from glycosylated 24 kDa hGH, collision-induced dissociation of tryptic glycopeptide T6 indicated that there had been an O-linked oligosaccharide attached to Thr-60. The sequence and branching structure of the oligosaccharide were determined by ESI-MS/MS analysis of tryptic glycopeptide T6. The mucin-like O-oligosaccharide sequence linked to Thr-60 begins with N-acetyl galactosamine and branches in a bifurcated topology with one appendage consisting of galactose followed by NeuAc and the other consisting of a single NeuAc. The oligosaccharide moiety lies in the high-affinity binding site 1 structural epitope of hGH that interfaces with both the growth hormone and the prolactin receptors and is predicted to sterically affect receptor interactions and alter the biological actions of hGH.

Health evaluation of free-ranging and captive pichis (Zaedyus pichiy; Mammalia, Dasypodidae), in Mendoza Province, Argentina.

The health of free-ranging and captive pichis (Zaedyus pichiy) was assessed in Mendoza Province, Argentina, between November 2001 and December 2006. Postmortem examinations of 150 confiscated and vehicle-killed pichis and clinical examinations of 139 wild-caught individuals suggest that the wild populations are currently in good health. Lesions and scars were observed in a large proportion of wild-caught pichis. The most common lesions were associated with parasitism or parasite larva migration. Sarcocystis cysts were relatively common in the skeletal muscle, and Besnoitia cysts were observed in the lungs of 24 evaluated animals. Elevated ambient humidity levels often caused moist dermatitis with epidermal detachment in captive pichis. This report constitutes the first health evaluation of free-ranging and captive Z. pichiy. It will be a starting point for future health studies and will be beneficial for the captive management of this species.

Respiratory disease associated with Bordetella bronchiseptica in a Hoffmann's two-toed sloth (Choloepus hoffmanni).

A 2-yr-old female captive-born Hoffmann's two-toed sloth (Choloepus hoffmanni) presented with respiratory disease. A severe inspiratory dyspnea with nasal congestion was observed with open-mouthed breathing and bilateral mucopurulent nasal exudate. Despite initial treatment with broad-spectrum antimicrobial therapy and anti-inflammatory and supportive care, the dyspnea persisted. The animal was anesthetized for bronchoscopy to obtain a deep tracheal sample. Based on culture of Bordetella bronchiseptica and sensitivity, a combination of systemic enrofloxacin, dexamethasone, and coupage with nebulization of saline, gentamicin, and albuterol as well as supportive care resulted in full recovery after 6 weeks of treatment.

Compressive myelopathy of the cervical spine in Komodo dragons (Varanus komodoensis).

Cervical subluxation and compressive myelopathy appears to be a cause of morbidity and mortality in captive Komodo dragons (Varanus komodoensis). Four cases of cervical subluxation resulting in nerve root compression or spinal cord compression were identified. Three were presumptively induced by trauma, and one had an unknown inciting cause. Two dragons exhibited signs of chronic instability. Cervical vertebrae affected included C1-C4. Clinical signs on presentation included ataxia, ambulatory paraparesis or tetraparesis to tetraplegia, depression to stupor, cervical scoliosis, and anorexia. Antemortem diagnosis of compression was only confirmed with magnetic resonance imaging or computed tomography. Treatment ranged from supportive care to attempted surgical decompression. All dragons died or were euthanatized, at 4 days to 12 mo postpresentation. Studies to define normal vertebral anatomy in the species are necessary to determine whether the pathology is linked to cervical malformation, resulting in ligament laxity, subsequent instability, and subluxation.

Lead poisoning and the reintroduction of the California condor in northern Arizona.

Since 1996, The Peregrine Fund has released California condors (Gymnogyps californianus) in the Grand Canyon region of northern Arizona with the goal of establishing a self-sustaining population, disjunct from other released populations in California and Baja California. A free-ranging population of more than 60 individuals now ranges within northern Arizona and southern Utah and has produced 9 wild young. The most frequent cause of death is lead poisoning from the ingestion of lead bullet fragments and shotgun pellets in the remains of gun-killed animals. In response, the Arizona Game and Fish Department has effectively reduced lead occurrence within the foraging range of the condors through hunter education and the promotion of nonlead ammunition. Most hunters have participated in the program. Throughout the course of the reintroduction effort, veterinary science and application have played essential roles in diagnosing fatalities and treating lead-exposed condors, a species with such a low natural reproductive rate that every adult is significant to the population.

DAPNe with micro-capillary separatory chemistry-coupled to MALDI-MS for the analysis of polar and non-polar lipid metabolism in one cell.

The cellular metabolome is considered to be a representation of cellular phenotype and cellular response to changes to internal or external events. Methods to expand the coverage of the expansive physiochemical properties that makeup the metabolome currently utilize multi-step extractions and chromatographic separations prior to chemical detection, leading to lengthy analysis times. In this study, a single-step procedure for the extraction and separation of a sample using a micro-capillary as a separatory funnel to achieve analyte partitioning within an organic/aqueous immiscible solvent system is described. The separated analytes are then spotted for MALDI-MS imaging and distribution ratios are calculated. Initially, the method is applied to standard mixtures for proof of partitioning. The extraction of an individual cell is non-reproducible; therefore, a broad chemical analysis of metabolites is necessary and will be illustrated with the one-cell analysis of a single Snu-5 gastric cancer cell taken from a cellular suspension. The method presented here shows a broad partitioning dynamic range as a single-step method for lipid analysis demonstrating a decrease in ion suppression often present in MALDI analysis of lipids. Graphical Abstract ᅟ.

FORCED MOLT IN FOUR JUVENILE YELLOW-EYED PENGUINS (MEGADYPTES ANTIPODES).

Penguins are dependent on waterproof plumage for survival. The molt in sub-Antarctic penguin species is a seasonal and catastrophic process during which the animals go through periods of fasting and high levels of stress. Their entire plumage is usually replaced in 3 wk. Attempts at consistent hormonal induction of molt in penguins have been unsuccessful. Four Yellow-eyed Penguins ( Megadyptes antipodes ) were referred for treatment at Wildbase, Massey University, Palmerston North, New Zealand, in late April 2014, following loss of waterproofing, feather breakage, increased body weight, pododermatitis, and damage to caudal feathers from hock sitting. Feather plucking of damaged areas to stimulate feather regrowth was attempted with poor results. Waiting 10-12 mo for a natural molt was not tenable. Catastrophic molt was induced by treatment with 10 g/kg of fresh beef thyroid gland orally once a day. The molt was complete in 18-26 d during which the animals regained full plumage and waterproofing after feather regrowth. The forced molt feathers had abnormal pigmentation but were of sufficient quality to allow release of the birds back to the wild.

Health evaluation of a radiocollared population of free-ranging Baird's tapirs (Tapirus bairdii) in Costa Rica.

The health of a population of free-ranging tapirs (Tapirus bairdii) inhabiting Corcovado National Park, Costa Rica, was assessed as part of an ongoing ecologic study. Nineteen tapirs were immobilized between March 1997 and February 2000, and samples of blood, skin biopsies, and ectoparasites were collected. Ticks were identified as Amblyomma oblongoguttatum or A. coelebs. Hematology and serum biochemistry results suggest statistically significant differences between free-ranging and captive populations that should be interpreted with caution in view of inherent environmental differences between the two populations. Five of 17 animals tested positive for Leptospira bratislava, and 12 individuals tested positive for Venezuelan equine encephalitis. One of nine skin biopsies examined was abnormal and diagnosed as leukoderma. This report represents the first health assessment of a free-ranging population of tapirs.

Population pharmacokinetics of antituberculous drugs and treatment of Mycobacterium bovis infection in bongo antelope (Tragelaphus eurycerus isaaci).

After clinical illness, treatment, and death of a captive male bongo antelope (Tragelaphus eurycerus isaaci) caused by tuberculosis involving Mycobacterium bovis, four tuberculin test reactive captive bongos were treated for 6 mo with isoniazid (INH) and rifampin (RIF) and intermittent single doses of other medications before being euthanized. In all cases, postmortem examination indicated no evidence of active disease and cultures of multiple organs were negative. We present detailed pharmacokinetic (PK) data for amikacin (AMK), ethambutol (EMB), INH, pyrazinamide (PZA), RIF, and levofloxacin in four female bongos. Adequate absorption and serum levels were obtained after parenteral administration of AMK, EMB, and INH and after oral administration of INH and PZA. Parenterally administered drugs were well described by a one-compartment PK model with first-order absorption and elimination processes. Treatment with INH and RIF over a 6-mo period did not result in demonstrable adverse effects. Starting doses of 10-15 mg/kg, i.m., or 30 mg/kg, p.o., of INH, 50 mg/kg, p.o., of EMB, and 25 mg/kg, i.m., s.i.d., of AMK are recommended. The treatment is continued with at least two drugs to which the organism is susceptible for a total treatment length of 6-12 mo. Treatment may be an option to eradicate M. bovis from suspect animals, with carefully administered and monitored drug treatment.

[Frequency and etiology of seminal infections in the study of infertile couples]. / Frecuencia y etiología de las infecciones seminales en el estudio de la pareja infértil.

UNLABELLED: Male genital infections are a relevant cause in the etiology of infertility, due to abnormalities in sperm quality, affecting spermatozoal count and motility. OBJECTIVE: Identify the most common bacteria isolated from seminal cultures and their impact on seminal quality. MATERIAL AND METHODS: A retrospective, longitudinal and observational study was performed by reviewing 295 files of infertile males seen in our clinic of Reproductive Biology at the Hospital Juárez de México, in whom seminogram and seminal cultures were practiced for detection of seminal infection and quality assessment. RESULTS: 47% of cases presented infection by one or two bacteria, Staphylococcus epidermidis, Streptococcus (alpha and beta hemolitic) and Escherichia coli prevailed. The main alterations in quality were motility, pH, morphology and viscosity in association with S. epidermidis and E. coli. CONCLUSIONS: Nearly half of our group of study disclosed seminal infections and it resembled the percentages reported in the literature; most of them were asymptomatic. In disparity with other reports the most common bacterial agent found in association with changes in seminal quality was a skin saprophyte.