Mapping genomic regions for reproductive traits in beef cattle: Inclusion of the X chromosome

Although the second largest chromosome of the genome, the X chromosome is usually excluded from genome-wide association studies (GWAS). Considering the presence and importance of genes on this chromosome that are involved in reproduction, the aim of this study was to evaluate the effect of its inclusion in GWAS on reproductive traits (scrotal circumference [SC], early pregnancy [P16] and age at first calving [AFC]) in a Nelore herd. Genotype data from 3,263 animals with the above-mentioned phenotypes were used. The results showed an increase in the variances explained by the autosomal markers for all traits when the X chromosome was not included. For SC, there was an increase of more than 10% for the windows on chromosomes 2 and 6. For P16, the effect was increased by almost 20% for windows on chromosome 5. The same pattern was found for AFC, with an increase of more than 10% for the most important windows. The results indicate that the noninclusion of the X chromosome can overestimate the effects of autosomes on SC, P16 and AFC not only because of the additive effect of the X chromosome itself but also because of its epistatic effect on autosomal genes.     

Treatment of experimental ureteral strictures by endourological ureterotomy and implantation of stents in the porcine animal model

The objective of this study is to evaluate the dilation of the ureter using endoureterotomy and an expanding-sheath double pigtail ureteral stent in the treatment of experimentally induced ureteral strictures in the porcine animal model. This is a new treatment in the ureteral strictures resolution in Veterinary Urology, although it is not a common affection, it usually appears as a consequence of ureteritis and in the iatrogenic female genital surgery. The experimental study is design in three phases: induction of experimental stricture, diagnosis and treatment of the stricture and follow-up. We have used 10 healthy Large White female pigs. The internal ureteral diameter was measured prior to laparoscopic ligature stricture induction using retrograde ureteropyelography (RUPG). Experimental stricture was diagnosed 4 weeks after intervention, using RUPG and ultrasound, and treated by endoureterotomy and subsequent placement of a double pigtail ureteral stent, which was removed 6 weeks later. The study finished 4 weeks later with measurement of ureteral diameters using RPUG and ultrasound evaluation. Except in one case, all ureters displayed permanent dilation of the strictured area for 10 weeks after treatment (6 weeks with ureteral stent and 4 more weeks without stent). Finally, this technique proved to be effective in cases of short-length and short-living ureteral strictures, and represents a viable alternative to conventional surgery in animals.     

Ultrastructural pathogenesis of the PRRS virus

Porcine Reproductive and Respiratory Syndrome Virus (PRRSV) was first isolated in swine alveolar macrophages (SAMs) and has subsequently been reported to replicate in other cell lines. Entry of the virus inside the cell takes place by receptor-mediated endocytosis. Following the entry of the virus into the cell, several not completely understood changes take place. PRRSV has been reported to be an apoptotic-inductor virus both in vivo and in vitro. Interestingly, it has been suggested that PRRSV-induced apoptosis occurs in cells other than those in which PRRSV replicates by a bystander mechanism. In this paper the ultrastructural pathogenesis of PRRSV will be reviewed.     

Morphometric analysis of the corneal endothelium of Yacare caiman (Caiman yacare) using scanning electron microscopy

The objective of this study was to examine the endothelial surface morphology and to perform morphometric analysis of the corneal endothelial cells of Yacare caiman (Caiman yacare) using scanning electron microscopy. Morphometric analysis with regard to polygonality, mean cell area, cell density and coefficient of variation of mean cell area was performed. Cell areas were measured using image analysis software. The normal corneal endothelium of Yacare caiman consisted of polygonal cells of uniform size and shape with interdigitations of the cell borders. Microvilli appeared as protrusions on the cellular surface. The average cell area was 270 +/- 24 microm(2) and the endothelial cell density was 3704 +/- 324 cells/mm(2). The coefficient of variation of cell area was 0.22. This study demonstrates that the Yacare caiman corneal endothelium is similar to those described in other vertebrates.     

Investigation of field outbreaks of turkey haemorrhagic enteritis in Hungary

Epidemiological, pathological, serological and virological investigations are reported on turkey haemorrhagic enteritis virus (THEV) infection in Hungarian turkey flocks. The pathogenesis of infection in experimentally infected turkeys and chickens, as well as the usefulness of polymerase chain reaction (PCR)/sequencing method for epidemiological investigation and for the differentiation of vaccine and field strains of THEV was also studied. Since the first recognition of the disease in Hungary in the late 1970s, until recently the disease has been diagnosed sporadically in its mild form. In the last few years (2000-2005), however, the number of outbreaks and the severity of the disease increased (9-23 affected flocks/year). Most of the outbreaks occurred at the age of 6 to 8 weeks and was complicated with Escherichia coli infection. The antibody levels to THEV in turkey flocks gradually declined till 5-7 weeks of age, and then they increased sharply due to natural infection with THEV. The immune response to vaccination (at 5 weeks of age) showed no significant antibody level increase one week postvaccination, but four weeks later the antibody level reached high values and then remained at this high level. The agar gel immunodiffusion (AGID) test to detect turkey adenovirus A (TAdV-A) antigen and PCR methods for THEV-specific DNA gave similarly positive results if spleens with pathognomonic lesions were tested; however, PCR proved to be more sensitive in cases with less characteristic pathological lesions. Nucleotide sequence alignment of PCR products amplified from Hungarian field strains and the Domermuth vaccine strain and that of the published THEV hexon sequences in GenBank database revealed slight differences between the sequences.     

Teratological examination of the insecticide methylparathion (Wofatox 50 EC) on pheasant embryos. 1. Morphological study

Teratogenic effects of methylparathion / Wofatox 50 EC/ and Parathion 20 WP as positive control material were tested in pheasant embryos. On the 12th day of incubation 0.1 ml of the emulsions or suspensions of the insecticides at different concentrations was inoculated into the air space of embryonated eggs. The following dose levels were employed: 27 and 270 mg/kg egg of Wofatox 50 EC, and 10 mg/kg of Parathion 20 WP. Morphological changes were evaluated by macroscopic, skeletal staining and light microscopic examinations of the embryos. Primary hypoplasia or atrophy developed in the cervical musculature /m. longus colli/ accompanied by lordosis and scoliosis of the cervical spine. In most cases we also found cyllosis .     

Acute toxoplasmosis in squirrel monkeys (Saimiri sciureus) in Mexico

Toxoplasma gondii causes fatal multisystemic disease in New World primates, with respiratory failure and multifocal necrotic lesions. Although cases and outbreaks of toxoplasmosis have been described, there are few genotyping studies and none has included parasite load quantification. In this article, we describe two cases of lethal acute toxoplasmosis in squirrel monkeys (Saimiri sciureus) of Mexico city. The main pathological findings included pulmonary edema, interstitial pneumonia, hepatitis and necrotizing lymphadenitis, and structures similar to T. gondii tachyzoites observed by histopathology in these organs. Diagnosis was confirmed by immunohistochemistry, transmission electron microscopy and both end point and real time PCR. The load was between <14 and 23 parasites/mg tissue. Digestion of the SAG3 gene amplicon showed similar bands to type I reference strains. These are the first cases of toxoplasmosis in primates studied in Mexico, with clinical features similar to others reported in Israel and French Guiana, although apparently caused by a different T. gondii variant.     

Analysis of the effect of the bovine adenosine triphosphate-binding cassette transporter G2 single nucleotide polymorphism Y581S on transcellular transport of veterinary drugs using new cell culture models

In commercial dairy production, the risk of drug residues and environmental pollutants in milk from ruminants has become an outstanding problem. One of the main determinants of active drug secretion into milk is the ATP-binding cassette transporter G2/breast cancer resistance protein (ABCG2/BCRP). It is located in several organs associated with drug absorption, metabolism, and excretion, and its expression is highly induced during lactation in the mammary gland of ruminants, mice, and humans. As a consequence, potential contamination of milk could expose suckling infants to xenotoxins. In cows, a SNP for this protein affecting quality and quantity of milk production has been described previously (Y581S). In this study, our main purpose was to determine whether this polymorphism has an effect on transcellular transport of veterinary drugs because this could alter substrate pharmacokinetics and milk residues. We stably expressed the wild-type bovine ABCG2 and the Y581S variant in Madin-Darby canine kidney epithelial cells (MDCKII) and MEF3.8 cell lines generating cell models in which the functionality of the bovine transporter could be addressed. Functional studies confirmed the greater functional activity in mitoxantrone accumulation assays for the Y581S variant with a greater relative V(MAX) value (P = 0.040) and showed for the first time that the Y581S variant presents greater transcellular transport of the model ABCG2 substrate nitrofurantoin (P = 0.024) and of 3 veterinary antibiotics, the fluoroquinolone agents enrofloxacin (P = 0.035), danofloxacin (P = 0.001), and difloxacin (P = 0.008), identified as new substrates of the bovine ABCG2. In addition, the inhibitory effect of the macrocyclic lactone ivermectin on the activity of wild-type bovine ABCG2 and the Y581S variant was also confirmed, showing a greater inhibitory potency on the wild-type protein at all the concentrations tested (5 μM, P = 0.017; 10 μM, P = 0.001; 25 μM, P = 0.008; and 50 μM, P = 0.003). Differential transport activity depending on the genotype together with the differential inhibition pattern might have clinical consequences, including changes in substrate pharmacokinetics (and subsequently pharmacodynamics) and more specifically, changes in secretion of ABCG2 substrates into milk, potentially implying important consequences to veterinary therapeutics.     

PCR-sequence characterization of new adenoviruses found in reptiles and the first successful isolation of a lizard adenovirus

A consensus nested PCR was used to screen diagnostic samples from approximately 70 reptiles for the presence of adenoviruses (AdV) in the years 2006-2007. Classical virus isolation methods were also used with all samples. After adenoviruses were detected in a group of helodermatid lizards in a Danish zoo, a follow-up study was also carried out on lizards from this group (10 Mexican beaded lizards and 24 Gila monsters) over the period of a year. Adenoviruses were detected in a total of 26 lizards and snakes by PCR. The PCR amplicons from all positive animals were sequenced and the resulting polymerase gene sequences were used for phylogenetic analysis. Altogether six Agamid AdVs were amplified, with a minimal sequence variation between one another and between these and GenBank Agamid AdVs. The sequence obtained from one of the Gila monsters is identical with the GenBank Helodermatid AdV, while the sequences from the Mexican beaded lizards differ from this. In a snake collection we have detected a new AdV from an Asp viper. All of the above mentioned adenoviruses cluster in the Atadenovirus genus. However, the sequence from a new Varanid AdV detected in this study clusters outside this genus. On cell culture, viruses were isolated from three of the AdV positive helodermatid lizards (one Mexican beaded lizard and two Gila monsters) and identified as AdVs based on electron microscopy and PCR and sequencing using cell culture supernatant. This is the first report of the successful isolation of a lizard AdV.