Increased incidence of DNA amplification in follicular than in uterine and blood samples indicates possible tropism of Neospora caninum to the ovarian follicle

This study evaluated the presence of Neospora caninum in ovarian follicle aspirates and uterine flushes obtained from N. caninum seropositive dairy cows. Ninety-two cows that aborted within the previous 90 days were sampled to determine the presence of antibodies against N. caninum. Thirteen seropositive cows were chosen for collection of blood leukocytes, uterine flushes (UF; n=12) and follicular aspirates (OPU; n=13). Samples were centrifuged and the cellular sediment from the follicular fluid, uterine flushes and blood leukocytes were used for DNA extraction and PCR. Follicular aspirates had the highest frequency of DNA amplification for N. caninum (p<0.05, 92.3%; 12/13). Whereas uterine (4/12) and blood leukocyte (5/13) samples had similar (p>0.05) rate of positive results. Nonetheless, there was no agreement between blood leukocytes and follicular samples taken from the same animal (Cohen Kappa=-0.16). Similarly, blood leukocytes and uterine results had moderate agreement between them (Cohen Kappa=0.47). This study indicates that N. caninum is present in the ovarian follicle and uterus of seropositive cows, suggesting a possible risk of neosporosis transmission between females during oocyte and embryo collection and transfer. Hence, precautions should be taken to minimize the risk of N. caninum transmission. Furthermore, the high incidence of positive results in follicle samples, that exceeded that of their paired blood leukocytes, suggests a possible tropism of N. caninum for the ovarian follicle.     

Effect of yeast products supplementation during transition period on metabolic profile and milk production in dairy cows

Tropical Animal Health and Production - The aim of the current study was to assess the effect that supplementation with yeast culture plus enzymatically hydrolyzed yeast (YC-EHY) during the...     

Effect of human tubal fluid medium and hyperactivation inducers on stallion sperm capacitation and hyperactivation

Conventional in vitro fertilization has not yet been implemented in the equine species. One of the main reasons has been the inability to develop a culture medium and incubation conditions supporting high levels of stallion sperm capacitation and hyperactivation in vitro. Although different culture media have been used for this purpose, human tubal fluid (HTF) medium, widely used in the manipulation of human and mice gametes, has not been reported so far in stallion sperm culture. The first part of this study aimed to compare HTF and Whitten's media on different stallion sperm quality and capacitation variables. Additionally, the effect of procaine, aminopyridine and caffeine in both media was evaluated on sperm motility parameters at different incubation times. Integrity and destabilization of the plasma membrane were evaluated by merocyanine 540/SYTOX Green (MC540), mitochondrial membrane potential (?Ψm) using tetramethylrhodamine methyl ester perchlorate (TMRM), acrosome membrane integrity by PNA/FITC and tyrosine phosphorylation by P‐tyrosine mouse mAb conjugated to Alexa Fluor® by flow cytometry. Motility parameters were evaluated using the integrated semen analysis system (ISAS®). We found no differences between Whitten's and HTF media and incubation time in terms of sperm viability, uninduced acrosome membrane damage or mitochondrial membrane potential at 30‐ and 120‐min incubation. Membrane fluidity (MC540) increased in both media at 30‐ and 120‐min incubation compared to noncapacitating conditions. Similarly, tyrosine phosphorylation increased in both media in capacitating conditions at 2‐ and 4‐hr incubation compared to noncapacitating conditions. Although procaine showed the best result in terms of sperm hyperactivated motility in both media, aminopyridine also showed parameters consistent with the hyperactivation including an increase in curvilinear velocity and decrease in straightness. In conclusion, HTF medium and aminopyridine equally support capacitation‐related parameters in stallion sperm.     

Encapsulated nitrate replacing soybean meal changes in vitro ruminal fermentation and methane production in diets differing in concentrate to forage ratio

The objective of this study was to evaluate the effects of using encapsulated nitrate product (ENP) replacing soybean meal in diets differing in concentrate to forage ratio on ruminal fermentation and methane production in vitro using a semi‐automatic gas production technique. Eight treatments were used in a randomized complete design with a 2 × 4 factorial arrangement: two diet (20C:80F and 80C:20F concentrate to forage ratio) and four levels of ENP addition (0%, 1.5%, 3.0%, and 4.5% of DM) replacing soybean meal. There was a diet × ENP interaction (p = 0.02) for methane production. According to ENP addition, diets with 80C:20F showed more intense reduction on methane production that 20C:80F. A negative linear effect was observed for propionate production with ENP addition in diet with 80C:20F and to the relative abundance of methanogens Archaea, in both diet. The replacement of soybean meal by ENP in levels up to 3% of DM inhibited methane production due to a reduction in the methanogens community without affecting the organic matter degradability. However, ENP at 4.5% of DM level affected fiber degradability, abundance of cellulolytic bacteria, and propionic acid production, indicating that this level of inclusion is not recommended for ruminant production.     

Detection of pseudocowpox virus in water buffalo (Bubalus bubalis) with vesicular disease in the state of São Paulo,Brazil, in 2016

Background: Parapoxviruses are zoonotic viruses that infect cattle, goats and sheep; there have also been reports of infections in camels, domestic cats and seals.

Objective: The objective of this report was to describe a case of vesicular disease caused by pseudocowpox virus (PCPV) in water buffalo (Bubalus bubalis) in Brazil.

Animals: Sixty buffalo less than 6 months old exhibited ulcers and widespread peeling of the tongue epithelium. There were no cases of vesicular disease in pigs or horses on the same property.

Methods: Samples were analysed by PCR and sequencing. Phylogenetic analysis in MEGA 7.01 was reconstructed using major envelope protein (B2L) by the Tamura three-parameter nucleotide substitution model and the maximum likelihood and neighbor joining models, both with 1000 bootstrap replicates. The genetic distance between the groups was analysed in MEGA using the maximum composite likelihood model. The rate variation among sites was modeled using gamma distribution.

Results: The presence of PCPV in the buffalo herd could be demonstrated in epithelium and serum. The minimum genetic distance between the isolated PCPV strain (262-2016) and orf virus and bovine papular stomatitis virus was 6.7% and 18.4%, respectively. The maximum genetic distance calculated was 4.6% when compared with a PCPV detected in a camel.

Conclusions/Clinical Importance: The peculiar position of the isolated strain in the phylogenetic trees does not necessarily indicate a different kind of PCPV that infects buffalo. More samples from cattle and buffalo in Brazil must be sequenced and compared to verify if PCPV from buffalo are genetically different from samples derived from cattle.     

Unraveling avian and reptilian hematology: An optical and electron microscopic study of the buffy coat

Background

Blood centrifugation and buffy coats are at the cornerstone of hematology. In mammals, the buffy coat has a layered disposition (from bottom to top) with neutrophils on top of erythrocytes, followed by monocytes/lymphocytes, and platelets. In nonmammals, this distribution is unknown. Recently, the cell tube block (CTB) technique was developed to study the buffy coat, but it was never applied to nonmammal buffy coats.

Objectives

This study aimed to evaluate using the CTB technique to study reptilian and avian buffy coats and to propose its use for clinical applications.

Methods

Blood from five birds and eight reptiles of different species was obtained to make CTBs that were processed for optical/electron microscopy. H&E, Sirius red, and immunohistochemistry staining against CD3 (to label T lymphocytes) were applied to the CTBs.

Results

In birds, the buffy coat had a layered appearance with the granulocyte layer containing granulocytes (heterophils and eosinophils) and nucleated erythrocytes followed by a mononuclear cell layer containing lymphocytes, monocytes, and thrombocytes. In some animals, a nucleated erythrocyte layer was observed admixed with the granulocyte/mononuclear cell layer. A small clot within the buffy coat was seen in seven reptiles, and less definition of layers occurred in reptiles, with only one or two layers. Lymphocytes appeared toward the top of the buffy coat.

Conclusions

From a comparative hematology perspective, the buffy coat of mammals differs from that of birds and more from that of reptiles. The CTB technique can be used to study these differences in avian and reptilian hematology, especially to study atypical circulating cells, hemoparasites, or blood cell proportions in health and disease.     

Pilot Study Investigating Fecal Shedding of Salmonella spp. by Horses Used in Competition

This pilot study was performed to determine the need for future investigation of the effect of transportation, environmental changes, and athletic competition on fecal shedding of Salmonella spp. in sport horses. Fecal samples were collected once daily from 20 healthy horses that were competing in National Collegiate Athletic Association equestrian event that occurred over a 3-day period. The horses were divided into four groups with five horses per group. The groups were divided based on travel distance. Fresh fecal samples were collected daily and cultured for Salmonella spp., through bacterial culture, for a minimum of 3 days for three groups that traveled to compete and for 15 days for a group that was housed at the competition location. All fecal samples were negative for Salmonella spp. Although this was a pilot study with a small sample size and limited collection days, the results of this study question the effects of the stress of transportation, new environment, and athletic activity on fecal shedding of Salmonella spp. in horses. A larger study, however, is needed to further investigate these findings.     

Management of the traumatic oronasal fistula in the cat with a conical silastic prosthetic device

Management of a traumatic oronasal fistula with a prosthetic device in an 8-year-old male neutered domestic shorthair cat is described. The animal had fallen from a height of three stories at the age of 1 year. The fall had resulted in a fracture of the hard palate and a split in the palatal mucosa. Two surgical attempts to close the defect had been unsuccessful and an autopolymerizable acrylic resin prosthesis had produced unsatisfactory results. The management of the chronic traumatic fistula with a conical prosthetic device was fast, easy and efficient. The device was removed and a second one was put in place after 2 years. This method would appear particularly useful for obturation of large palatal defects that have failed to heal after attempts at surgical closure and is an alternative treatment for debilitated cats which may not be candidates for longer surgical procedures.