Development of an indirect ELISA-NcSRS2 for detection of Neospora caninum antibodies in cattle

Neosporosis is of alarming economic concern in the cattle industry. The effectiveness of diagnostic tests for detecting specific antibodies against Neospora caninum is hampered by potential cross-reaction with other coccidia. Use of a single specific antigen might improve test specificity. An indirect enzyme-linked immunosorbent assay (ELISA) was developed using the truncated protein NcSRS2 expressed in Escherichia coli. The ELISA results were compared with those of the indirect fluorescence antibody test (IFAT). Receiver Operating Characteristic (ROC) and Tests in the Absence of a Gold Standard (TAGS) analysis revealed an assay having 96% specificity and 95% sensitivity when applied to 145 positive and 352 negative sera from two distinct cattle populations. Using OD ≤ 0.095 as the cut-off point, the assay's negative and positive predictive values ranged from 98.8% to 50.8% and from 58.8% to 99.1%, respectively, depending on neosporosis prevalence in a given area. The novel ELISA-NcSRS2 format described in the present report constitutes a specific and sensitive method for detecting N. caninum in cattle.     

Oocysts of Hepatozoon canis in Rhipicephalus (Boophilus) microplus collected from a naturally infected dog

Canine hepatozoonosis is a tick-borne disease caused by protozoans of the genus Hepatozoon. Several tick species have been implicated as potential vectors. Therefore, extensive studies are needed to determine the 'natural' endemic cycle of this parasite. This paper presents the first report of the presence of Hepatozoon canis oocysts in Rhipicephalus (Boophilus) microplus collected from an infected dog.     

In vitro development of IVF-derived bovine embryos following cytoplasmic microinjection for the episomal expression of the IGF2 gene

Important genomic imprinting changes usually occur following the in vitro production (IVP) of bovine embryos, especially in the imprinting pattern of components of the IGF system. This study aimed to evaluate the effects of a transient episomal overexpression of the IGF2 gene in bovine IVP embryos following embryo cytoplasmic microinjection (CMI) at the 1-cell stage on embryo survival, early and late developmental kinetics and morphological quality up to Day 7 of development. Selected cumulus–oocyte complexes (COCs) were matured and fertilized in vitro and subsequently segregated into six experimental groups: non-CMI control group and five CMI groups at increasing doses (0, 10, 20, 40 and 80 ng/μl) of a GFP vector built for the episomal expression of bovine IGF2. Zygote CMI was effective in delivering the expression vector into the ooplasm, irrespective of the groups, with 58% of positive GFP fluorescence in Day 7 blastocysts. Considering developmental rates and late embryo kinetics, the 10-ng/μl CMI vector dose promoted a lower blastocyst rate (10.4%), but for blastocysts at more advanced stages of development (93.0% blastocysts and expanded blastocysts), and higher number of cells (116.0 ± 3.0) than non-CMI controls (23.3%, 75.0% and 75.0 ± 6.8 were obtained, respectively). In conclusion, CMI at the 1-cell stage did not compromise subsequent in vitro development of surviving embryos, with the 10-ng/μl group demonstrating a possible growth-promoting effect of the IGF2 gene on embryo development, from the 1-cell to the blastocyst stage.     

Radiographic anatomy of the cockatiel (Nymphicus hollandicus) axial and appendicular skeleton

Cockatiels are popular pets. Still, despite medical and surgical relevance, the radiographic anatomy of the cockatiel (Nymphicus hollandicus) skeleton, like that of different wild and exotic bird species, has seldom been described. This study set out to describe the radiographic anatomy of the cockatiel skeleton. Twelve adult male and nine adult female specimens were radiographed using a digital X-ray system and different views. The radiographic anatomy of these birds was similar to that of other Psittacidae. However, some particularities inherent to the target species were detected, such as the presence of four flexion zones in the skull (craniofacial, nasal, jugal arch and palatine), complete bony orbit comprising a suborbital arch, 34–38 vertebrae (10 or 11 cervical, 8 or 9 thoracic, 9 or 10 lumbosacral, 5 or 6 caudal vertebrae and a pygostyle comprising 2 fused vertebrae), eight or nine pairs of ribs and a notarium made up of fused T2–T6 vertebrae. Poor radiopacity of the notarium, ribs and respective uncinate processes, and synsacral vertebrae made demarcation of these structures difficult. The appendicular skeleton of the cockatiel was very similar to that of other Psittacidae, and there were no gender-related differences.     

Quantitative and descriptive histological aspects of jaguar (Panthera onca Linnaeus, 1758) ear skin as a step towards formation of biobanks

Skin of mammals vulnerable to extinction, such as the jaguar, is used as a source of material in conservation strategies. The composition of skin is not uniform among species, and the ability to distinguish similarities in skin morphology in animal groups is fundamental in the application of skin tissue for use in biobanks. The aim of our study was to evaluate the structure, composition and capacity for culture of ear skin from the yellow and black jaguars. Both qualitative and quantitative methods were used, focusing on skin thickness, cell quantification and distribution, collagen density, proliferative activity and viability. Histomorphometrical study of the skin showed a total thickness of 273.2 and 274.6 µm for the yellow and black jaguars, respectively. Melanocytes and fibroblasts were, respectively, 9.7 and 23.0 for the yellow jaguar and 11.3 and 26.8 for the black jaguar. A collagen density of 67.0% and 49.0% was observed for yellow and black jaguars, respectively. Both animals presented a proliferative activity varying between 1.20 and 1.30. All tissues could promote cellular detachment, reaching subconfluence in 10–15 days. This kind of information from histomorphometrical features and cell cultures can be essential for a more targeted application of ear skin cryopreservation in this species, as such information will enable understanding the action of substances on tissues during the conservation process.     

Digit innervation of the thoracic limb of Criollo horses: Anatomical description and consequences to perineural blocks

Criollo horse breeding is an important economic activity in South America. Because of their athletic performance, these animals tend to show great incidence of musculoskeletal disorders, many of them diagnosed by means of perineural blocks. However, incorrect interpretation of these blocks may be due to anatomical differences in nerve distribution. The objective of this study was to describe the innervation of the digit region of thoracic limbs in Criollo horses, in order to improve the interpretation of tests for claudication diagnosis based on nerve block. Thirty thoracic limbs from Criollo horses were dissected. It could be observed that in 90% of the limbs, dorsal branches of the palmar nerve originated proximally to the proximal sesamoid bone. In 93% of the cases, the palmar digital nerve and the dorsal branches communicated; in 87% of the cases, communication between branches of the dorsal branch was observed; and in 27% (8/30) of the limbs, the palmar metacarpal nerve and the dorsal branch presented communications. None of the specimens showed complete symmetry in the distribution of nerves in contralateral limbs. The high frequency of communication between the nerves may be a particularity of the Criollo breed that may interfere with the interpretation of perineural blocks. Based on the anatomical position, it may be inferred that divergent results in Criollo horses may occur when abaxial sesamoid nerve block is used. Palmar digital nerve block may be less influenced by these variations, provided it is performed as distal as possible from the ungular cartilage.     

Removal of Metaldehyde Through Photocatalytic Reactions Using Nano-Sized Zinc Oxide Composites

In this study, we have reported the synthesis of new amino-substituted p-tert-butylcalix[4]arene (3) and its application for the removal of two carcinogenic azo dyes, i.e., Chicago Sky Blue (CSB) and Tropaeolin 000 (TP) from aqueous environment. The newly synthesized calix–ligand 3 is characterized by FT-IR and ¹H NMR spectroscopy as well as elemental analysis. The extraction efficiency of newly calix–ligand 3 for CSB and TP dyes from aqueous media was evaluated through liquid–liquid extraction experiments. The newly synthesized calix–ligand 3 showed outstanding extraction percentage and maximum percent extraction, i.e., 97 and 96 % of CSB and TP dyes was achieved at pH 9, respectively. During the extraction process, effect of various parameters was monitored and found that extraction is highly dependent on pH and salinity. Moreover, cyclic structure, cavity size, functional groups of the calixarene derivative, hydrophobicity, and the ionic property of guest molecules also affect the extraction efficiency. The comparative data prop up calix–ligand 3 as an effective extractant for both CSB and TP dyes.     

Punctodera achalensis n. sp. (Nematoda: Punctoderidae), a new cyst nematode from Argentina

European Journal of Plant Pathology - Trichoderma aggressivum, a mycopathogen causing green mould disease, is a major problem in Agaricus bisporus cultivation due to crop loss, and resistance to...