Acaricidal properties of the essential oil and precocene II obtained from Calea serrata (Asteraceae) on the cattle tick Rhipicephalus (Boophilus) microplus (Acari: Ixodidae)

Calea serrata Less. (Asteraceae), an endemic species of south Brazil known as "quebra-tudo", is used in Afro-Brazilian religious rituals and in folk medicine for treating liver disorders. Phytochemical studies of the n-hexane extract of this plant demonstrated the presence of precocene II, a benzopyran derivative known for its insecticidal activity. The aim of this work was to isolate this benzopyran and determine the chemical composition of the essential oil of C. serrata and further to evaluate the acaricidal activity of the essential oil and precocene II against the larvae of Rhipicephalus (Boophilus) microplus. The LC(99.9) and LC(50) values obtained with the oil, which presents precocene II and sesquiterpenes, were 3.94 μL/mL and 0.28 μL/mL, respectively. For precocene II this values were 4.25mg/mL and 1.78 mg/mL, respectively. The results indicate a synergistic interaction between the components of the oil and precocene II.     

Priapism Secondary to Perineal Abscess in a Dog – A Case Report

A 7‐year‐old intact male Boxer was referred to our services at the Veterinary Teaching Hospital of the University of Trás‐os‐Montes and Alto Douro, suffering from a persistently erect penis (including the bulbus glandis) that had been exposed for several days. Radiographic and ultrasonographic examinations detected a 5.0 × 3.5 cm mass located dorso‐laterally to the urinary bladder. The microbial culture of the mass revealed Staphylococcus spp. At that time, we suspected the involvement of an abscess in the origin of the priapism. Medical and surgical treatments were promptly instituted, which allowed for penile withdrawal into the prepuce; however, the resolution of the penile erection was not accomplished in the following days and penile amputation was required. Histological evaluation of the excised penis revealed extensive infarction of the erectile tissue of the pars longa and bulbus glandis, and also of the blood vessels of the penis. Following penile amputation and antimicrobial therapy, the animal fully recovered. Ultimately, the animal died as a consequence of gastric torsion. At necropsy, some lesions compatible with a previous perforation of the intestinal wall were recorded. The data gathered from the anamnesis, the physical and imaging examinations, along with the post‐mortem findings, allowed us to conclude that in this clinical case the primary cause of priapism was a perineal abscess due to bowel perforation.     

Response of Thawed Epidi dymal Red Deer Spermatozoa to Increasing Concentrations of Hydrogen Peroxide,and Importance of Individual Male Variability

Oxidative stress represents a challenge during sperm manipulation. We have tested the effect of increasing hydrogen peroxide (H₂O₂) levels on red deer spermatozoa after cryopreservation, and the role of male‐to‐male variation in that response. In a first experiment, eight thawed samples were submitted to 0, 25, 50, 100 and 200 μm H₂O₂ for 2 h at 37°C. Intracellular reactive oxygen species (H₂DCFDA‐CM) increased with H₂O₂ concentration, but we only detected a decrease in sperm function (motility by CASA and chromatin damage by sperm chromatin structure assay) with 200 μm . Lipoperoxidation assessed by the thiobarbituric acid reactive substance (TBARS) method increased slightly with 50 μm H₂O₂ and above. In a second experiment, samples from seven males were submitted to 0 and 200 μm H₂O₂ for 2 h, triplicating the experiment within each male. Males differed at thawing and regarding their response to incubation and H₂O₂ presence. We found that the kinematic parameters reflected male‐to‐male variability, whereas the response of the different males was similar for lipid peroxidation and viability. A multiparametric analysis showed that males grouped differently if samples were assessed after thawing, after incubation without H₂O₂ or after incubation with H₂O₂. Red deer spermatozoa are relatively resilient to H₂O₂ after thawing, but it seems to be a great male‐to‐male variability regarding the response to oxidative stress. The acknowledgement of this individual variability might improve the development of optimized sperm work protocols.     

Using BCG,MPT-51 and Ag85 as antigens in an indirect ELISA for the diagnosis of bovine tuberculosis

This study evaluated the Mycobacterium tuberculosis protein antigen MPT-51, the trimeric antigen 85 (Ag85) complex, and Bacillus Calmette-Guérin (BCG) in an indirect ELISA to diagnose bovine tuberculosis (TB) from serum samples. Serum was collected from 208 intra-dermal tuberculin test (ITT)-positive and 54 ITT-negative animals from a region where bovine TB is endemic. Using the Ag85 and BCG antigens, the indirect ELISA was able to discriminate ITT-positive from ITT-negative animals. This level of discrimination was not achieved when using the MPT-51 antigen. The highest sensitivity (Se) and specificity (Sp) of the test was found when BCG was used (Se, 82%; Sp, 91%). Further work in different epidemiological settings and with larger numbers of animals will be required to validate these findings.     

Genotyping of potentially zoonotic Giardia duodenalis from exotic and wild animals kept in captivity in Brazil

We have studied the variability of glutamate dehydrogenase (gdh) and small subunit ribosomal (SSU) rRNA coding genes of Giardia species in fecal samples isolated from wild and exotic animals in Brazil, and compared with homologous sequences of isolates from human and domestic animals characterized in previous studies. Cysts of Giardia duodenalis were obtained from feces of naturally infected monkeys (Alouatta fusca) (n=20), chinchillas (Chinchilla lanigera) (n=3), ostriches (Struthio camelus) (n=2) and jaguar (Panthera onca) (n=1). Assemblage AI was assigned to the unique isolate of jaguar. All the samples from monkeys, chinchillas, and ostriches were assigned to Assemblage B. There was little evolutionary divergence between the referred isolates and isolates described elsewhere. The Assemblage B isolates identified in this study were closely related to Assemblage BIV isolated from humans. The molecular identification of Assemblages A and B of G. duodenalis isolates from exotic and wild animals demonstrates that such hosts may be a potential reservoir for zoonotic transmission of G. duodenalis.     

Bovine Oocyte Quality in Relation to Ultrastructural Characteristics of Zona Pellucida,Polyspermic Penetration and Developmental Competence

In the present study, the effect of bovine oocyte quality related to ultrastructural characteristics of zona pellucida (ZP), polyspermic penetration and embryo developmental competence was evaluated. Cumulus–oocyte complexes were punctioned from 453 ovaries, classified as 1, 2, 3 and 4 according to their morphological aspect, matured for 24 h and then divided into two groups. In group A, oocytes were fixed in 2.5% glutaraldehyde and 0.1 m sodium cacodylate and examined under a scanning electron microscope. Photomicrographs were taken and ZP’s pores were evaluated in squares of 6.4‐μm width. In group B, oocytes were fertilized in vitro. After 48 h, non‐cleaved oocytes were fixed for polyspermy evaluation. On days 7, 9 and 10, embryos were classified as developed (blastocysts and hatched blastocysts). Results showed that quality 1 oocytes revealed a ZP pore diameter of 0.50 ± 0.07 μm, which was smaller than the observed on oocytes of quality 2 (0.83 ± 0.10 μm), quality 3 (1.02 ± 0.22 μm) and quality 4 (1.38 ± 0.59 μm) (p ≤ 0.05). For In Vitro Fertilization (IVF), results showed that embryos originating from oocytes classed as 3 and 4 had lower cleavage rate (68.4% and 43.8%) than those belonging to class 1 and 2 (79.5% and 69.3%) (p ≤ 0.05). None oocyte classified as 3 and 4 developed to hatch blastocysts, while for oocytes belonging to quality 1 and 2, these values were, respectively, 15.2% and 12.5%. Concerning polyspermy, oocytes class 1 and 2 had lower polyspermic penetration than those belonging to class 3 and 4 (respectively 4.1%, 4.5%, 11.1% and 9.8%, for class 1, 2, 3 and 4). In conclusion, the present study demonstrated that oocytes with low qualities result in lower developmental competence and with high percentage of polyspermy after IVF, which can be the result of the ZP structure such as the number and the pore’s diameter.     

Influence of different supplements and sugarcane (Saccharum officinarum L.) cultivars on intake, digestible variables and methane production of dairy heifers under tropical conditions

The sulphur hexafluoride (SF(6)) gas tracer method was used to measure methane (CH(4)) production of crossbred (3/4 Holstein x Zebu) dairy heifers fed two types of sugarcane (Saccharum officinarum L.; cultivar IAC-862480 (CC1) or cultivar IAC-873184 (CC2)) and supplemented with urea or concentrate. The study was performed at Embrapa Southeast Cattle, S?o Carlos, SP, Brazil, using a completely randomised design. Differences between treatments were significant for digestibility of dry matter, organic matter and energy. When animals were supplemented with urea differences between sugarcane cultivars did occur for NDF consumption, but not for daily methane production. This suggest that variation in chemical composition of sugarcane did not affect bovine ruminal CH(4) emissions. Concentrate inclusion in animal diet increased digestible organic matter intake, improving the nutrient intake by animals, but did not reduce CH(4) production expressed as a percentage of gross energy intake.     

Trans-10, cis-12 conjugated linoleic acid and the PPAR-γ agonist rosiglitazone attenuate lipopolysaccharide-induced TNF-α production by bovine immune cells

Lipopolysaccharide (LPS) modulates innate immunity through alteration of cytokine production by immune cells. The objective of this study was to examine the effect of exogenous conjugated linoleic acid (CLA) and PPAR-γ agonist, rosiglitazone, on LPS-induced tumor necrosis factor α (TNF-α) production by cultured whole blood from prepubertal Holstein heifers (mean age, 5.5 mo). Compared with unstimulated cells, addition of LPS (10 μg/mL) to the culture medium increased (P < 0.03) peripheral blood mononuclear cell proliferation ≤2.5-fold. Coincubation with interferon γ (5 ng/mL) further stimulated (P < 0.01) the lymphoproliferative response to LPS. Lipopolysaccharide increased (P < 0.01) TNF-α concentration in cultured whole blood in a dose- and time-dependent manner. The greatest TNF-α stimulation occurred after 12 h of exposure to 1 μg/mL LPS. Coincubation with trans-10, cis-12 CLA isomer (100 μM) or rosiglitazone (10 μM), a PPAR-γ agonist, decreased (P < 0.01) LPS-induced TNF-α production by 13% and 29%, respectively. Linoleic acid and cis-9, trans-11 CLA isomer had no detectable effects on LPS-induced TNF-α production in cultured bovine blood. The PPAR-γ agonist-induced TNF-α attenuation was reversed when blood was treated with both rosiglitazone and GW9662, a selective PPAR-γ antagonist. Addition of rosiglitazone to the culture medium tended to reduce nuclear factor-κ Bp65 concentration in nuclear and cytosolic extracts isolated from cultured peripheral blood mononuclear cells. Results show that LPS is a potent inducer of TNF-α production in bovine blood cells and that trans-10, cis-12 CLA and PPAR-γ agonists may attenuate the pro-inflammatory response induced by LPS in growing dairy heifers. Additional studies are needed to fully characterize the involvement of nuclear factor-κ B in LPS signaling in bovine blood cells.     

Surgical Repair of an Inguinal Herniation of the Urinary Bladder in an Intact Female Domestic Rabbit (Oryctolagus cuniculus)

A 4-month-old intact female domestic rabbit (Oryctolagus cuniculus) was evaluated for a recent onset of lethargy, anorexia, foul-smelling red urine, diarrhea, and a soft subcutaneous swelling in the inguinal region. Physical examination confirmed the presence of a flocculent subcutaneous swelling in the ventral pelvic region. Whole body radiographs showed an approximately 5 × 3.5 × 3.5 cm structure of mineral opacity within the subcutaneous tissues ventral to the pelvic girdle. Cytology of fluid aspirated from the mass and an excretory urogram supported the diagnosis of a herniation of the urinary bladder through the right caudoventral abdominal body wall. The hernia was successfully repaired with an inguinal herniorrhaphy and replacement of the bladder within the abdominal cavity. The etiology of this herniation could not be definitively determined but was likely congenital or non-traumatically acquired in origin. This is the first published report describing the surgical repair of an inguinal herniation and incarceration of the urinary bladder in an intact female rabbit.