Identification of Hammondia heydorni oocysts by a heminested-PCR (hnPCR-AP10) based on the H. heydorni RAPD fragment AP10

Toxoplasma gondii, Hammondia hammondi, Neospora caninum, Neospora hughesi and Hammondia heydorni are members of the Toxoplasmatinae sub-family. They are closely related coccidians with similarly sized oocysts. Molecular diagnostic techniques, especially those based on polymerase chain reaction (PCR), can be successfully applied for the differentiation of Hammondia-like oocysts. In this paper, we describe a rapid and simple method for the identification of H. heydorni oocysts among other members of the Toxoplasmatinae sub-family, using a heminested-PCR (hnPCR-AP10) based on a H. heydorni RAPD fragment available in molecular database. DNA of oocysts of H. heydorni yielded a specific fragment of 289-290 bp in the heminested-PCR assay. No product was yielded when the primers were used for the amplification of DNA extracted from T. gondii, N. caninum, N. hughesi and H. hammondi, thus allowing the differentiation of H. heydorni among other members of the Toxoplasmatinae sub-family. The hnPCR-AP10 was capable of detecting H. heydorni genetic sequences from suspensions with at least 10 oocysts. In conclusion, the hnPCR-AP10 proved to be a reliable method to be used in the identification of H. heydorni oocysts from feces of dogs.     

Transmission electron microscopy for characterization of acrosomal damage after Percoll gradient centrifugation of cryopreserved bovine spermatozoa

The objective of this study was to characterize acrosomal ultrastructure following discontinuous Percoll gradient centrifugation of cryopreserved bovine sperm. Semen was collected from six bulls of different breeds and three ejaculates per bull were evaluated. Frozen semen samples were thawed and the acrosomal region of sperm cells was evaluated by transmission electron microscopy (TEM) before (n = 18) and after (n = 18) Percoll centrifugation. The evaluation of 20 sperm heads from each of the 36 samples analyzed ensured that a large number of cells were investigated. The data were subjected to analysis of variance at a level of significance of 5%. Percoll centrifugation reduced the percentage of sperm exhibiting normal acrosomes (from 61.77 to 30.24%), reduced the percentage of sperm presenting atypical acrosome reactions (from 28.38 to 4.84%) and increased the percentage of sperm exhibiting damage in the acrosome (from 6.14 to 64.26%). The percentage of sperm with typical acrosome reactions was not significantly different before (3.70%) and after (0.67%) centrifugation. TEM distinguished four different types of acrosomal status and enabled ultrastructural characterization of acrosomal injuries. The percentage of sperm exhibiting normal acrosomes decreased and damage in the acrosome was the most frequent acrosomal injury with the Percoll gradient centrifugation protocol utilized.     

Amyloidosis in Captive Anseriformes

During the period 1945-1962, percentage deaths of anserine birds (ducks, geese and swans) did not vary remarkably from year to year even though the population has more than quadrupled during the same period. The number of deaths with amyloidosis has increased significantly even though the death rate did not change. The number of deaths due to tuberculosis, mould, renal diseases, hepatitis and infection accompanied by amyloidosis also increased, especially during the second half the study period. Cases of known secondary amyloidosis accounted for 71.4% whereas 28.6% of the remaining cases were classified as ”doubtfully” primary. Sites of deposition amyloid in the two categories were the same and it has never been observed in the heart. The amyloid deposits in the birds did not differ from those in man when examined by number of histochemical methods.     

Length of productive life of sows in four pig farms in the tropics of Mexico

The objective of this study was to determine the effect of some factors on the length of productive life (LPL), the number of parities (NP), and the culling interval (CI) of sows. The information from 2003 to 2009 of 3,746 sows with LPL and NP records and 5,819 sows with CI records from the four farms of Yucatan, Mexico was used. LPL was defined as the number of days between the first farrowing and culling or death and CI as the number of days between the last farrowing and culling. To determine the effects of farm, litter size at first farrowing (LSF1) and age at first farrowing (AF1) on LPL and NP, and of farm, LSF1 and culling reason on CI general linear models were used. Means and standard errors for LPL, NP, and CI were 496.4 ± 8.42 days, 3.35 ± 0.04 parities, and 48.3 ± 0.52 days, respectively. All factors had significant effects (P < 0.05) on LPL, NP, and CI. The sows from farm 2 stayed longer (806.0 days) and had more parities (5.25) than the sows from the other farms. Sows with the smallest litters (≤8 piglets) at first farrowing stayed the least time in the farm (675.1 days) and had the fewest parities (4.49). Gilts farrowing the first time before 331 days of age stayed longer time in the farm (726.4 days) and had more parities (4.81) in comparison with 331–348 and more than 348 days age groups. With respect to CI, the sows in farm 3 stayed the longest time in the farm. Sows with more than five parities were culled sooner after their last farrowing. In conclusion, differences between farms for LPL, PN, and CI were found. Females that farrowed at a short age and those with large litters stayed longer in the herd. Old sows and those culled for reproductive reasons stayed the least time in the herd after their last farrowing.     

A multiplex PCR for the detection of Brucella spp. and Leptospira spp. DNA from aborted bovine fetuses

Bovine brucellosis and leptospirosis are important causes of bovine abortion around the world. Both diseases can be serologically diagnosed, but many factors may cause false positive and negative results. Direct methods based on bacteriological isolation are usually employed, but they are difficult, time consuming and dangerous. Monoplex polymerase chain reaction (PCR) have been successfully described for the detection of Brucella spp. and Leptospira spp. Aiming at improvement in the direct diagnosis, a multiplex PCR (mPCR) for the detection of these agents in aborted bovine fetuses is described. The detection threshold of the mPCR was evaluated in experimentally contaminated bovine clinical samples using a conventional proteinase K/SDS or a boiling-based extraction protocols. The mPCR was applied to two groups of clinical samples: 63 episodes of bovine abortion and eight hamsters experimentally infected with Leptospira interrogans serovar pomona. Adopting microbiological isolation as reference, the test showed a sensitivity of 100% in both groups of clinical samples. Seven samples collected from bovine fetuses were Brucella spp. culture negative but showed positive results in mPCR. Regarding Leptospira spp. detection, similar results were observed in three bovine clinical samples. All hamsters infected with Leptospira were positive in both microbiological culture and mPCR. The boiling extraction protocol showed better results in some clinical samples, probably by the removal of PCR inhibitors by heat treatment. The high sensitivity, simplicity and the possibility of detection of both bacteria in a single tube reaction support the use of the mPCR described in the routine diagnosis.     

Hyaluronidase administered with xylazine–tiletamine–zolazepam into adipose tissue shortens recovery from anesthesia in pigs

Objective

To evaluate the effect of hyaluronidase on uptake, duration and speed of elimination of xylazine–tiletamine–zolazepam administered in the subcutaneous fat over the dorsal lumbar region of swine.

Pulse pressure variation as a guide for volume expansion in dogs undergoing orthopedic surgery

Objective

To investigate whether pulse pressure variation (PPV) can predict fluid responsiveness in healthy dogs during clinical surgery.

Development and validation of a radioimmunoassay for thyrotropin in cattle.

In mammals, thyrotropin, or thyroid-stimulating hormone (TSH), assay is used for the diagnosis of primary hypothyroidism. Hypothyroidism is the most common type of thyroid disorder in cattle. The aim of this study was to develop and validate, under physiologic and pathologic conditions, a radioimmunoassay (RIA) for bovine TSH (bTSH). Double RIA was performed with purified bTSH and specific bovine antiserum. Laboratory validation included research of minimal detection limit, accuracy, and reproducibility. The physiologic validation included a thyrotropin-releasing hormone (TRH) challenge performed on euthyroid cows and a follow-up of bTSH concentration over a 24-hour period. Furthermore, bTSH concentration was assayed in a large population of healthy dairy and beef cows to define reference interval. The pathologic validation was made by assaying bTSH and thyroid hormones on healthy and goitrous newborn calves. The minimum detection limit (MDL) for bTSH assay was 1.3 microU/ml. The recovery was 101% to 106%. The intra- and interassay coefficients of variation (CVs) ranged from 5% to 11% and 11% to 15%, respectively. The RIA covered the whole range of physiologic bTSH values, as shown by bTSH values induced by TRH-challenge. A pulsatile secretion of bTSH was observed, accompanied by a diurnal variation with lower night values than day values. Reference intervals of bTSH ranged from 1.3 to 13.0 microU/ml for beef and dairy breeds. Finally, bTSH easily discriminated goitrous newborn calves from healthy ones, leading to the definition of a cutoff value of 35 microU/ml. The bTSH assay positively reacted to physiologic and pathologic conditions. The accuracy and precision of the RIA were satisfying.