Photodynamic therapy for the treatment of periocular squamous cell carcinoma in horses: a pilot study

Objective  Local photodynamic therapy (PDT) is a novel cancer therapy in veterinary ophthalmology. A prospective pilot study seeking to demonstrate proof of principle and safety for the treatment of equine periocular squamous cell carcinoma (PSCC) was therefore conducted. We hypothesized that surgical excision with adjunctive local PDT is an effective and safe treatment for equine PSCC.
Procedures  Nine horses (10 eyes) with PSCC were treated with surgical resection, local infiltration of resulting wound beds with 2-[1-hexyloxyethyl]-2-devinylpyropheophorbide-a (HPPH) and irradiation with 665-nm wavelength diode laser. Regular follow-up ophthalmic examinations were performed.
Results  Surgical resection and PDT yielded disease-free intervals of 25–68 months in our study horses as of January, 2008. These results were obtained following a single treatment in seven horses and two treatments in one horse. In one horse, carcinoma in situ developed 2.5 months after partial surgical excision and PDT, requiring local excision under standing sedation.
Conclusions  Preliminary results suggest that surgical resection and adjunctive local PDT is a safe and effective novel treatment for PSCC in horses. More research is needed before PDT for the treatment of equine PSCC can be adequately compared with other current modalities. Important to future investigations regarding PDT, tumor recurrence rate, length of hospitalization, number of treatment episodes required to effect tumor remission, and total treatment costs should be examined in a controlled manner. Our present results and experiences suggest that this treatment may be useful in the treatment of equine PSCC.     

A comparison of the uterine proteome of mares in oestrus and dioestrus

Proteomic analysis of mare uterine flush fluid provides a minimally invasive technique for studying protein changes associated with the oestrous cycle. The aim of this study was to identify differentially abundant proteins in the uterine flush fluid of mares in oestrus and dioestrus. In this study, uterine flush fluid samples were collected from eight reproductively healthy mares in either oestrus (n = 5) or dioestrus (n = 3). Proteomic analysis was performed using liquid chromatography‐tandem mass spectrometry. Of 172 proteins identified, six proteins (immunoglobulin lambda‐like polypeptide 1, haemoglobin subunit alpha, alpha‐1B‐glycoprotein, serotransferrin, apolipoprotein A‐1, and haemoglobin subunit beta) were significantly more abundant in oestrus. These proteins may contribute to the endometrial defence system through roles in inflammation, immunity or antimicrobial activity. In other species, some of these proteins have been described as immunoglobulins, negative acute phase proteins or defence agents against micro‐organisms. During dioestrus, immunoglobulin alpha‐1 chain C region‐related, complement factor I, CD 109 antigen and uterocalin, were significantly more abundant. Research in other species suggests that these four proteins contribute to the immune response through proposed immunoregulatory characteristics, complement system involvement or roles in B cell–T cell interactions. In conclusion, ten differentially abundant proteins were identified in the uterine flush fluid of mares in oestrus and dioestrus. Targeted studies on these proteins could elucidate their role in uterine defence mechanisms during the oestrous cycle in the mare.     

Application of different pharmacokinetic models to describe and predict pharmacokinetics of voriconazole in magellanic penguins following oral administration

Aspergillosis is a condition causing serious morbidity and mortality in captive penguins and other bird species. It can be treated with antifungal drugs, such as voriconazole. However, the pharmacokinetics of voriconazole are variable between different animal and bird species. Therefore, the pharmacokinetics of voriconazole were investigated in this study in Magellanic penguins. Pharmacokinetic models were constructed and applied to predict the pharmacokinetics of voriconazole during long‐term treatment in Magellanic penguins, since the voriconazole treatment duration in chronic aspergillosis cases can last up to several months. Plasma voriconazole concentration–time data from adult Magellanic penguins (Spheniscus magellanicus; n = 15) following a single oral (PO) dose of either 2.5 mg/kg or 5 mg/kg in a herring in three separate study periods 7–12 months apart were collected. Mean plasma voriconazole concentrations were above the targeted MIC for Aspergillus fumigatus for 2 hr following a single 2.5 mg/kg voriconazole dose while the plasma concentrations exceeded the MIC for least 24 hr following a 5 mg/kg dose. Nonlinear mixed‐effects modeling was used to fit two pharmacokinetic models, one with first‐order and another with saturable elimination, to the single‐dose data. Fits were good for both, as long as dose was included as a covariate for the first‐order model so that clearance was lower and the half‐life longer for animals receiving the 5 mg/kg dose. Although the single‐dose data suggested saturated elimination at higher concentrations, the model with saturable elimination did not predict plasma voriconazole concentrations well for a clinical aspergillosis case receiving long‐term treatment, possibly because of induction of metabolizing enzymes with chronic exposure. Pharmacokinetic models should accurately predict plasma drug concentrations for different dosage regimens in order to be applicable in the field. Future studies should focus on determining clearance at steady‐state to be able to refine the pharmacokinetic models presented here and improve model performance for long‐term oral voriconazole administration in Magellanic penguins.     

Role of sugars in surface microbe-host interactions and immune reaction modulation

Sugars in the form of monosaccharides, oligosaccharides, polysaccharides and glycoconjugates (glycoproteins, glycolipids) are vital components of infecting microbes and host cells, and are involved in cell signalling associated with modulation of inflammation in all integumental structures. Indeed, sugars are the molecules most commonly involved in cell recognition and communication. In skin, they are essential to epidermal development and homeostasis. They play important roles in microbial adherence, colonization and biofilm formation, and in virulence. Two groups of pathogen recognition receptors, C-type lectins (CTL) and their receptors (CTLR), and the Toll-like receptors enable the host to recognize pathogen-associated molecular patterns (PAMPs), which are mainly glycolipids. The CTLs can recognize a wide variety of bacteria, fungi and parasites and are important in phagocytosis and endocytosis. TLRs are expressed on the surfaces of a variety of cells, including keratinocytes, dendritic cells, monocytes and macrophages; they play a major role in innate immunity. Interaction of TLRs with PAMPs initiates a cascade of events leading to production of reactive oxygen intermediates, cytokines and chemokines, and promotes inflammation. Exogenous sugars can block carbohydrate receptors and competitively displace bacteria from attachment to cells, including keratinocytes. Thus sugars may provide valuable adjunctive anti-inflammatory and/or antimicrobial treatment. A promising approach is the use of a panel of carbohydrate derivatives with anti-adhesive efficacy against bacteria frequently involved in diseases affecting skin and other epithelia. More complete characterization of sugar receptors and their ligands will provide further keys to use of carbohydrates in immunomodulation and infection control in skin.     

Detection of QTL for forage yield,lodging resistance and spring vigor traits in alfalfa (Medicago sativa L.)

Alfalfa (Medicago sativa L.) is an internationally significant forage crop. Forage yield, lodging resistance and spring vigor are important agronomic traits conditioned by quantitative genetic and environmental effects. The objective of this study was to identify quantitative trait loci (QTL) and molecular markers associated with increased forage yield, resistance to lodging, and spring vigor. A backcross population composed of 128 progeny was developed by crossing the breeding parents DW000577 (lodging susceptible) and NL002724 (lodging-resistant) and back-crossing an individual F₁ plant to the maternal parent (i.e. DW000577). A linkage map of NL002724 was developed based upon the segregation of 236 AFLP, SRAP, and SSR markers among the backcross progeny. The markers were distributed among 14 linkage groups, covering an estimated recombination distance of 1497.6 centiMorgans (cM). Replicated clones of both parents and backcross progeny were evaluated in the field for estimated forage yield, lodging, and spring vigor in Washington and Wisconsin during 2007 and 2008. Significant QTL were found for all three traits. In particular, two QTL for lodging resistance were identified that explained ≥14 % of trait variation, and were significant in all years and locations. Major QTL explaining over 25 % of trait variation for forage yield were detected in multiple environments at two separate locations on chromosome III. Several QTL for spring vigor were located in the same or similar positions as QTL for forage yield, possibly explaining the significant correlation between these traits. Molecular markers associated with the aforementioned QTL were also identified.     

Reduced effective population size in an overexploited population of the Nile crocodile (Crocodylus niloticus)

Unchecked exploitation of wildlife resources is one of the major factors influencing species persistence throughout the world today. A significant consequence of exploitation is the increasing rate at which genetic diversity is lost as populations decline. Recent studies suggest that life history traits affecting population growth, particularly in long-lived species, may act to moderate the impact of population decline on genetic variation and lead to remnant populations that appear genetically diverse despite having passed through substantial demographic bottlenecks. In this study we show that the retention of genetic variation in a partially recovered population of Nile crocodile is deceptive, as it masks the reality of a significant decline in the population’s effective size (N_e). Repeated episodes of unchecked hunting in the mid to late 20th century have today led to a five-fold decrease in the population’s N_e. Using current census data we estimate the contemporary N_e/N ratio as 0.05 and, in light of quotas that permit the ongoing removal of adults, simulated the likely effects of genetic drift on extant levels of variation. Results indicate that even if the current effective size is maintained, both allelic diversity and heterozygosity will decline. Our findings have complex implications for long-lived species; an emphasis on the retention of genetic variation alone, whilst disregarding the effects of population decline on effective size, may ultimately obscure the continued decline and extinction of exploited populations.     

In vitro susceptibility patterns of Aspergillus and Fusarium species isolated from equine ulcerative keratomycosis cases in the midwestern and southern United States with inclusion of the new antifungal agent voriconazole

Objective  To evaluate and compare the in vitro susceptibility of Aspergillus and Fusarium spp. isolated from horses with ulcerative keratomycosis, address regional differences in equine keratomycosis isolates, and provide susceptibility data to update prior studies.
Animal studied  Fourteen horses with ulcerative keratomycosis.
Procedures  Banked fungal isolates from equine corneal ulcers (eight Aspergillus spp. and six Fusarium spp.) were identified at The University of Texas Health Science Center at San Antonio. In vitro minimum inhibitory concentration and susceptibility to natamycin, fluconazole, itraconazole, voriconazole, ketoconazole, and miconazole were determined for each isolate.
Results  Fungi were significantly more susceptible to voriconazole than to natamycin, itraconazole, fluconazole, and ketoconazole, but miconazole susceptibility did not differ significantly from voriconazole. Aspergillus spp. were most susceptible to voriconazole, miconazole, and itraconazole, which were significantly better to fluconazole and ketoconazole. Fusarium spp. susceptibility was greatest to natamycin and voriconazole and lowest to itraconazole and ketoconazole. Fusarium spp. were significantly less susceptible to itraconazole and ketoconazole compared to natamycin. No significant differences in susceptibility were found when isolates from Florida were compared with isolates from other states.
Conclusions and clinical relevance  Based on in vitro evidence, voriconazole appears to be the most effective antifungal for initial treatment of equine keratomycosis in the midwestern and southern United States. Results are comparable with previous studies in that isolated fungi from equine keratomycosis cases showed consistently poor susceptibility to fluconazole. Organisms isolated in different geographic locations of the midwestern and southern United States appeared to have similar patterns of antifungal susceptibility.     

Occurrence of Neospora caninum in dogs and its correlation with visceral leishmaniasis in the urban area of Campo Grande, Mato Grosso do Sul, Brazil

Neospora caninum is an obligate intracellular protozoan that can infect domestic and wild canids, as well as ruminants and equines, and is described as causing neuromuscular alteration and death in dogs. Visceral leishmaniasis (VL) is an infectious disease that affects both humans and animals, being caused by protozoan parasites of the genus Leishmania, of which Leishmania (Leishmania) chagasi is found in Brazil -- transmitted by sand flies, such as Lutzomyia longipalpis, in most of the American continent. The immunosuppression caused by VL can promote the occurrence of co-infections with other agents. In order to determine the frequency of N. caninum and its relationship to VL in Campo Grande, MS, Brazil, 345 blood sera were collected from dogs. The sera were submitted to an indirect fluorescent antibody test (IFAT) for detection of anti-N. caninum antibodies and VL antibodies. N. caninum was found in 26.5% of VL-negative dogs and in 29% of VL-positive ones. Among males, it was found in 30.7% of VL-negative animals and in 30.4% of VL-positive ones; among females, in 21% of VL-negative animals and in 27.7% of VL-positive ones. Among juvenile dogs (under 1 year), N. caninum was detected in 10.5% of VL-negative animals and in 11.2% of VL-positive ones. For adult dogs (1 year and older) the results were 31.4% for VL-negative animals and 28.8% of VL-positive ones. The study revealed a statistically significant association with age (chi(2)=9.76, P<0.05) in the N. caninum results for VL-negative animals. No significant correlation in N. caninum seroprevalence was found when VL-positive or VL-negative dogs were compared (chi(2)=0.21, P=0.64). The findings suggest that in Campo Grande N. caninum and VL co-infection is common in dogs, though VL does not appear to enhance susceptibility to N. caninum.     

Trawl fishing impacts on the status of seabed fauna in diverse regions of the globe

Bottom trawl fishing is a controversial activity. It yields about a quarter of the world's wild seafood, but also has impacts on the marine environment. Recent advances have quantified and improved understanding of large‐scale impacts of trawling on the seabed. However, such information needs to be coupled with distributions of benthic invertebrates (benthos) to assess whether these populations are being sustained under current trawling regimes. This study collated data from 13 diverse regions of the globe spanning four continents. Within each region, we combined trawl intensity distributions and predicted abundance distributions of benthos groups with impact and recovery parameters for taxonomic classes in a risk assessment model to estimate benthos status. The exposure of 220 predicted benthos‐group distributions to trawling intensity (as swept area ratio) ranged between 0% and 210% (mean = 37%) of abundance. However, benthos status, an indicator of the depleted abundance under chronic trawling pressure as a proportion of untrawled state, ranged between 0.86 and 1 (mean = 0.99), with 78% of benthos groups > 0.95. Mean benthos status was lowest in regions of Europe and Africa, and for taxonomic classes Bivalvia and Gastropoda. Our results demonstrate that while spatial overlap studies can help infer general patterns of potential risk, actual risks cannot be evaluated without using an assessment model that incorporates trawl impact and recovery metrics. These quantitative outputs are essential for sustainability assessments, and together with reference points and thresholds, can help managers ensure use of the marine environment is sustainable under the ecosystem approach to management.