Low‐density lipoproteins and milk serum proteins improve the quality of stallion sperm after vitrification in straws

Lipids and proteins can be used for sperm vitrification to preserve the integrity of sperm membranes or to increase the viscosity of the medium. This study evaluated the effect of low‐density lipoproteins (LDL) and milk serum proteins (Pronexcell) for stallion sperm vitrification. Hippex extender (Barex Biochemical Products, The Netherlands), plus 1% of bovine serum albumin and 100 mM of trehalose, was used as control for sperm vitrification. In experiment 1, different concentrations of LDL (L1 = 0.25, L2 = 0.5, L3 = 1%) and in experiment 2 of Pronexcell (P1 = 1, P2 = 5, P3 = 10%) were added to control extender. Vitrification was performed in 0.25‐ml straws directly plunged into liquid nitrogen. Total motility (TM, %) and progressive motility (PM, %) were analysed by CASA, and plasma membrane (IMS, %) and acrosome membrane integrity (AIS, %) were assessed under epifluorescence microscopy. Post‐warmed sperm parameters were compared between treatments by ANOVA. Results were expressed as mean ± SEM. In both experiments, the minimum concentration of LDL and Pronexcell obtained significantly higher values (p < 0.01) than the control extender for TM (L1 = 52.95 ± 4.4; P1 = 58.99 ± 4.6; C = 30.88 ± 3.0), PM (L1 = 36.79 ± 5.5; P1 = 47.25 ± 4.3; C = 19.20 ± 2.4), IMS (L1 = 68.88 ± 3.6; P1 = 47.25 ± 4.3; C = 52.81 ± 2.6) and AIS (L1 = 45.88 ± 3.6; P1 = 47.25 ± 4.3; C = 26.00 ± 2.1). No differences in sperm parameters were found among different concentrations of LDL or Pronexcell. In conclusion, the addition of 0.25% LDL and 1% Pronexcell to the vitrification extender is recommended to improve the quality of stallion sperm after vitrification.     

Morphometric and quantitative evaluation of the NADH-diaphorase positive myenteric neurons of the jejunum of streptozotocin-diabetic rats supplemented with acetyl-L-carnitine

Summary In this study we investigated the effect of the acetyl-L-carnitine (ALC) supplementation on the myenteric neurons of the jejunum of rats made diabetic at the age of 105 days by streptozotocin (35 mg/kg body weight). Four groups were used: non-diabetic (C), non-diabetic supplemented with ALC (CC), diabetic (D), diabetic supplemented with ALC (DC). After 15 weeks of diabetes induction the blood was collected by cardiac puncture to evaluate glycaemia and glycated haemoglobin. Next the animals were killed and the jejunum was collected and subjected to whole-mount preparation to evidence the myenteric neurons through the histochemical technique of the NADH-diaphorase. The neuronal counts were made in 80 microscopic fields, in tissue samples of five animals of each group. The profiles of the cell bodies of 1000 neurons per group were analysed. Diabetes induced a significant increase in the area of the cell body and decrease in the number of NADH-diaphorase positive myoenteric neurons. ALC suplementation to the diabetic group promoted smaller hypertrophic effects and less neuronal loss than in the myoenteric neurons of the diabetic rats, and in addition diminished the body weight decrease and reduced the fasting glycaemia.     

How to Perform and Interpret Testicular Fine Needle Aspiration in Stallions

Although several methods of testicular biopsy have been proposed previously, testicular fine needle aspiration (FNA) has proved to be the simplest, the most rapid, inexpensive, and overall the least invasive technique for obtaining testicular biopsies. Testicular FNA is indicated for fertility investigations in stallions with oligozoospermia or azoospermia. It is also used for differential diagnosis of testicular enlargement. After sedation, the stallion’s testis is punctured to obtain testicular parenchyma samples containing cells mainly from the seminiferous epithelium. The material obtained is used to perform smears which are analyzed for identification and quantification of germ cells and Sertoli cells. The results are based on the presence of the cell types found in the smears and the proportions of Sertoli cells per germ cells. In addition to being a very useful diagnostic tool, testicular FNA is also used for follow-up examinations, as it is minimally invasive.     

Brucellosis in Brazil

This paper reviews the epidemiology of bovine, swine, ovine, caprine, and canine brucellosis in Brazil. The zoonotic aspects of Brucella infection in Brazil is also discussed. Emphasis is given to the new program for the control of brucellosis in cattle and buffaloes that is likely to provide important insights into the prospects and strategies for controlling brucellosis in developing countries.     

In Vitro Assessment of the Equine Hoof Wall Strains in Flat Weight Bearing and After Heel Elevation

Horseshoeing is a common practice, but effects on the hoof wall are poorly understood. Strain gauges were used to document and compare hoof behavior in vitro during flat weight bearing and after artificial heel elevation. Ten front limbs of Thoroughbred race horses, shod with conventional flat shoes, were used. Eight strain gauges were symmetrically distributed around the toe, quarters, and heels. Each limb was mounted to a testing machine (Kratos K5002; Kratos Dynamômetros, Ltda., Cotia-SP-Brazil) and subjected to a load equivalent to 30% of the donor's body weight. Strains (μ) were acquired by means of a computerized system and the results compared using Friedman and Wilcoxon statistical tests. There was greater strain variation when the heels were elevated. Compression predominated during flat weight bearing, with a tendency to horizontal traction after heel elevation. The changes in strain caused by heel elevation were not always symmetrical. Elevation of the heels tensed the toe and the medial quarter horizontally, increased load at the posterior portion of the hoof capsule, and hindered its expansion.     

Effect of oral trazodone on the minimum alveolar concentration of isoflurane in dogs

Objective

To determine the effect of oral trazodone on the minimum alveolar concentration (MAC) of isoflurane in dogs.

Influence of murine Toxocara canis infection on plasma and bronchoalveolar lavage fluid eosinophil numbers and its correlation with cytokine levels

Toxocara canis is a nematode of the Ascaridae family that normally parasites the small intestine of canid species. Humans are accidentally infected upon ingestion of embryonated eggs, and can manifest several clinical alterations such as fever, hepatomegaly, splenomegaly, respiratory symptoms, muscle pain and anorexia. In the present work, we investigated the kinetics of tissue distribution of L2 larva in lungs, liver, kidney, brain, skeletal muscle and myocardium. Also, we analyzed the blood and bronchoalveolar lavage fluid (BAL) for levels of IL-6, IFN-gamma, eotaxin and Regulated on Activation Normal T Cell Expressed and Secreted (RANTES) in experimental murine T. canis infection. We observed liver, lung and kidney lesions correlated to larva migration as early as the first day of infection. After the seventh post-infection day, larva could also be detected in brain, skeletal muscle and heart, as an indicator of biphasic migration pattern. Increased inflammatory activity was detected in BAL and plasma of infected animals, as was an intense eosinophil migration associated with an increase in the levels of all the cytokines studied. In conclusion, our results establish a tight correlation between tissue lesions caused by larva migration and increased plasma levels of pro-inflammatory and eosinophil chemotactic cytokines. Thus, murine T. canis infection may prove to be useful in understanding the role of cytokines in infection.     

Different isolates from Leishmania braziliensis complex induce distinct histopathological features in a murine model of infection

The aim of this study was to evaluate the histopathological features in tissues of mice infected by human isolates (I, II, and III) or the reference M2903 strain of Leishmania braziliensis complex. BALB/c and C57Bl/6 mice were infected in the hind footpad with 10⁶ stationary-phase promastigotes of L. braziliensis complex. The evolution of lesions was observed for 10 weeks and the animals were then euthanized and liver, spleen and popliteal lymph nodes were collected. Tissues were stained with hematoxylin and eosin and analyzed by immunohistochemistry assay. Increased thickness of infected footpads was observed in all animals, lesions were nodular and non-ulcerated. Mice infected with isolate I presented inflammatory infiltrates consisting predominantly of mononuclear cells in all tissues examined, and also a great number of megakaryocytes, compared with other isolates. Infection with isolate II led to an infected footpad enlargement not seen in other isolates. In addition, mononuclear infiltrates in the liver and hemosiderin in spleen were noted. Conversely, mice infected with either isolate III or M2903 strain only showed an increased number of megakaryocytes in spleen. All tissues examined had detectable amastigote forms of Leishmania by immunohistochemistry in all groups. Taking together, our results showed an unforeseen behavior of different isolates of L. braziliensis complex that led to diverse pathological 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.