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.     

The effect of dietary supplementation with 9-cis:12-trans and 10-trans:12-cis conjugated linoleic acid (CLA) for nine months on serum cholesterol, lymphocyte proliferation and polymorphonuclear cells function in Beagle dogs

Conjugated linoleic acids (CLA), 9-cis:11-trans and 10-trans:12-cis, have been shown to be able to modify some immune cells parameters and plasma lipids in a variety of experiment models. Since lymphocytes and polymorphonuclear cells (PMNC) have a large spectrum functions in the immune response, the knowledge in this field has to be expanded. Beagle dogs were fed a control diet or a CLA supplemented diet for nine months. Blood was collected for biochemical analysis and lymphocyte and PMNC isolation. PMNC were assayed for lysosome content, phagocytic activity and superoxide anion production. A lymphocyte proliferation capacity assay was done. The CLA fed dogs had a 34% reduction in total cholesterol (P < 0.05), 28% in LDL (P < 0.05) and 28% non-HDL-cholesterol (P < 0.05). Neither of the PMNC parameters evaluated demonstrated significant alteration. Lymphocytes from CLA group increased by 45% their mitotic capacity (P < 0.05). Our study demonstrates that CLA can successfully modify the lipid profile of dogs (monogastrics) when fed at reasonable levels, but did not significantly alter inflammatory function as would generally predicted. Further, we had some indication that CLA modulated T cell responsiveness.     

The efficiency of energy retention of broiler chickens and turkeys fed on diets with different lysine concentrations

1.?An experiment determined the effect of 6 different lysine concentrations (40, 52, 58, 65, 90 and 130 g/kg CP) in two protein concentrations (210 and 260 g/kg) on the efficiency of energy utilisation of broiler chickens and turkeys. Ninety-six male Ross 308 broiler chicks and 96 male BUT Big 6 turkey poults were fed at 90% of ad libitum during a 12 d bioassay period. At the end of the bioassay, birds were slaughtered and carcase crude protein, fat and gross energy were measured and nutrient metabolisability determined.

2.?Broiler chickens had lower AME and AME_N than turkeys. AME was higher in the 260 g/kg than in the 210 g/kg protein diet; however, no difference was evident in AME_N between any of the diets in both species. A significant species × lysine interaction indicated that the decrease in AME and AME_N with increasing lysine was greater in turkeys than in broiler chickens.

3.?Turkeys had lower weight gains and efficiencies of energy retention than broiler chickens. The effects of different lysine concentrations on growth and the efficiency of energy retention were significant and growth responses were quadratic but there were negative linear responses in the efficiency of energy retention in both species at both crude protein concentrations. The efficiency of energy retention was increased with decreasing lysine concentrations throughout the wide range of concentrations used in the experiment.     

Estimation of genetic parameters for mature weight in Angus cattle

The aim of this study was to estimate genetic parameters for BW of Angus cattle up to 5 yr of age and to discuss options for including mature weight (MW) in their genetic evaluation. Data were obtained from the American Angus Association. Only records from herds with at least 500 animals and with >10% of animals with BW at ≥ 2 yr of age were considered. Traits were weaning weight (WW, n = 81,525), yearling weight (YW, n = 62,721), and BW measured from 2 to 5 yr of age (MW2, n = 15,927; MW3, n = 12,404; MW4, n = 9,805; MW5, n = 7,546). Genetic parameters were estimated using an AIREML algorithm with a multiple-trait animal model. Fixed effects were contemporary group and departure of the actual age from standard age (205, 365, 730, 1,095, 1,460, and 1,825 d of age for WW, YW, MW2, MW3, MW4, and MW5, respectively). Random effects were animal direct additive genetic, maternal additive genetic, maternal permanent environment, and residual. Estimates of direct genetic variances (kg(2)) were 298 ± 71.8, 563 ± 15.1, 925 ± 52.1, 1,221 ± 65.8, 1,406 ± 80.4, and 1,402 ± 66.9; maternal genetic variances were 167 ± 4.8, 153 ± 6.1, 123 ± 9.1, 136 ± 12.25, 167 ± 18.0, and 110 ± 14.0; maternal permanent environment variances were 124 ± 2.9, 120 ± 4.3, 61 ± 7.5, 69 ± 11.9, 103 ± 15.9, and 134 ± 35.2; and residual variances were 258 ± 3.8, 608 ± 8.6, 829 ± 34.2, 1,016 ± 38.8, 1,017 ± 52.1, and 1,202 ± 63.22 for WW, YW, MW2, MW3, MW4, and MW5, respectively. The direct genetic correlation between WW and YW was 0.84 ± 0.14 and between WW and MW ranged from 0.66 ± 0.06 (WW and MW4) to 0.72 ± 0.11 (WW and MW2). Direct genetic correlations ranged from 0.77 ± 0.08 (YW and MW5) to 0.85 ± 0.07 (YW and MW2) between YW and MW, and they were ≥ 0.95 among MW2, MW3, MW4, and MW5. Maternal genetic correlations between WW and YW and MW ranged from 0.52 ± 0.05 (WW and MW4) to 0.95 ± 0.07 (WW and YW), and among MW they ranged from 0.54 ± 0.14 (MW4 and MW5) to 0.94 ± 0.07 (MW2 and MW3). Genetic correlations suggest that a genetic evaluation for MW may be MW2-based and that including BW from older ages could be accomplished by adjusting records to the scale of MW2.     

Adjusted method of penis fixation during boar semi-automatic semen collection aiming to reduce bacterial contamination

Semen collection has an essential role in the initial bacterial load in boar ejaculates and extended semen. The study aimed to explore the efficacy of an adjusted penis fixation in a semi-automatic collection system on reducing bacterial contamination of ejaculates in two-boar studs with different scenarios. Historically, stud A had low levels of bacterial load in raw semen, while stud B had a high level of contamination. A total of 56 mature boars had their semen collected using two methods of penis fixation: (a) Traditional: The penis was fixed directly with the artificial cervix and transferred to the adjustable clamp; (b) Adjusted: The fixation was performed with one gloved-hand, and after exteriorization, the penis was gripped using the artificial cervix with the other gloved-hand and transferred to the adjustable clamp. The bacterial load (p = .0045) and the occurrence of ejaculates >231 CFU/ml (p = .0101) were reduced in the Adjusted compared to the Traditional method. Bacterial load was reduced when using the Adjusted method in stud B (p = .0011), which showed a greater occurrence of critical factors for bacterial contamination (p ≤ .0034). The Adjusted method reduced the occurrence of ejaculates >231 CFU/ml when the preputial ostium was dirty (p = .016) and the duration of semen collection was >7 min (p = .022) compared to the Traditional method. In conclusion, the Adjusted penis fixation was efficient in reducing bacterial load of ejaculates, mainly in boar stud B, which had high contamination challenges.     

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.     

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.     

Non-accidental injuries found in necropsies of domestic cats: a review of 191 cases

Animal cruelty is defined as a deliberate action that causes pain and suffering to an animal. In Brazil, legislation known as the Environmental Crimes Law states that cruelty toward all animal species is criminal in nature. From 644 domestic cats necropsied between January 1998 and December 2009, 191 (29.66%) presented lesions highly suggestive of animal cruelty. The main necroscopic finding was exogenous carbamate poisoning (75.39%) followed by blunt-force trauma (21.99%). Cats from 7 months to 2 years of age were the most affected (50.79%). In Brazil, violence is a public health problem and there is a high prevalence of domestic violence. Therefore, even if laws provide for animal welfare and protection, animals are common targets for violent acts. Within a context of social violence, cruelty toward animals is an important parameter to be considered, and the non-accidental lesions that were found are evidence of malicious actions.     

Platelet‐Activating Factor and Evidence of Oxidative Stress in the Bronchoalveolar Fluid of Thoroughbred Colts during Race Training

Background: Inflammatory airway disease (IAD) is prevalent in young racehorses during training, being the 2nd most commonly diagnosed ailment interrupting training of 2‐year‐old Thoroughbred racehorses. Hypothesis: That stabling and exercise cause oxidative stress, release of platelet‐activating factor (PAF) and inflammation in airways of Thoroughbred colts. Animals: Colts in breeding farms (NC, n = 45), stabled for 30 days (EC, n = 40), and race trained (EX, n = 34). Methods: Cytological profile and parameters of bronchoalveolar lavage fluid (BALF) related to oxidative stress, bioactivity of the proinflammatory mediator PAF, catalase activity, and alveolar macrophage function. Results: Percentages of neutrophils and eosinophils in the BALF of the EX group were higher (5.4 ± 6.4% versus 0.9 ± 1.2%) than the upper limits for normal horses (3–5%). BALF from the EX group (45.6 ± 2.8 cells/μL of BALF) also displayed significantly (P= .017) higher total nucleated cell count. PAF bioactivity and the total protein concentration in the BALF were higher in the EX group (0.0683 ± 0.076 versus 0.0056 ± 0.007 340 : 380 nm ratio P= .0039, 0.36 ± 0.30 versus 0.14 ± 0.15 mg of proteins/mL of BALF P < .001). Concentration of BALF hydroperoxides was higher in the EC group (104.7 ± 80.0 versus 35.2 ± 28.0 nmol/mg of proteins, P= .013) and catalase activity was higher in the EX group (0.24 ± 0.16 versus 0.06 ± 0.02 μmol H₂O₂/min/mg of proteins, P= .0021). Alveolar macrophage phagocytosis (P= .048) as well as production of superoxide anion (P= .0014) and hydrogen peroxide (P= .0011) were significantly lower in EX group. Conclusions and Clinical Importance: Further studies should be performed to elucidate the role of PAF in the pathophysiology of IAD. Its presence in bronchoalveolar fluid of young athletic horses makes it a potential therapeutic target to be investigated.     

Disseminated pythiosis in three horses

Three cases of equine subcutaneous pythiosis with dissemination to the internal organs were investigated. The subcutaneous lesions were observed on the mammary gland, nostrils and limbs of the infected horses. Histopathological analysis of the infected tissues revealed a strong eosinophilic reaction, with macrophages, mast cells and giant cells. Sparsely septated hyphal filaments of 4-6 microm diameter were identified in the center of the eosinophilic areas. Specific fluorescent antibody against Pythium insidiosum confirmed the hyphae in the infected tissues in three examined horses. In one of the three cases, the DNA sequences amplified from the infected subcutaneous tissues and internal organs, revealed that P. insidiosum's 18S SSU rDNA amplicons shared 100% identity with those sequences deposit in GenBank. This is the first report confirming by immunochemical and genetic techniques that P. insidiosum can disseminated from superficial to deep structures.